DE2356862A1 - 1-Oxa 6-amino-penicillanic acid deriv. analogues - including antibacterials, prepd. in several stages starting with anhydropenicillin - Google Patents

Abstract

Cpds of formulae (I) and (II), cationic salts of (I) and pharmacologically acceptable non-toxic salts of (II) are new. (where R1CO is acyl). (II) are antibacterials used as 5-200 (pref. 5-20) mg/kg/day (I) are intermediates for (I). Specif claimed (II) are those in which R is benzyl, phenoxymethyl, 2,6-dimethoxyphenyl, alpha-methoxy-3,4-dichlorobenzyl, alpha-acetoxybenzyl or alpha-aminobenzyl.

Description

PROF, DR. DR. J. REITSTÖTTER

DR.-ING. WOLFRAM BUNTE. ; DR. WERNER KINZEBACH

O - βΟΟΟ MUNICH 4O. BAUERSTRA88K ZZ · FERNRUF (ΟΘ0) 37 ββ θ »■ TELEX * 02102O8 ISAR D POSTANSCHRIFT« D-βΟΟΟ MUNICH 43. POST BOX 7SO

Munich, the 7th new animal in 1974 M / 12860.

Queen's University RRl, Kingston, Ontario / CANADA. ',

Oxygen analogs of 6-aminopenicillanic acid derivatives and

Process for their manufacture

The invention relates to oxygen analogs of 6-aminopenicillanic acid derivatives and methods of making them.

The process begins with the chlorination of various types of anhydropenicillins, producing sulfur-free dichlorides that react with water or alcohols, whereby acids or esters are formed. These esters still contain one chlorine atom, which is a slight intermolecular one

1"

509820/1161

nucleophilic displacement by azide, cyanide, acetate or Formations, or subject to intramolecular displacement, forming oxazolines, oxazolidones and oxa-anhydropenicillins will. Allyl bromination of these monocyclic and bicyclic azetidinones followed by reaction with a Azide ion results in an allylic azide group that can be reduced to an allylic amino group. Most of the intermediates produced in this way are new. From these Compounds are new ß-lactam antibiotics, such as 1-0xahsnzylpenicillin, manufactured.

Penicillins and cephalosporins comprise a group of well-known antibacterial agents which are generally grouped into a class called β-lactam antibiotics. JP Hou and JW Poole, ß-Lactam-Antibiotics: Their Physicochemical Properties and Biological Activities in Relation to Structure, J. Pharmaceutical Sciences, 60 ( .4) , 503, 532 (April, 1971) * Most of the work in this area has, generally speaking, been undertaken essentially with 6-aminopenicillanic acid, 7-aminocephalosporanic acid and derivatives prepared therefrom by fermentation.

Considerable work has also been done on total chemical synthesis. The text provides a more recent overview by M.S. Mannas and A.K, Böse, Synthesis of Penicillin, Cephalosporin C and Analogues, Marcel Decker, Inc., 95 Madison Avenue, New York, New York 1969. An even more recent one R.B. Morin and B.G. Jackson, Chemistry of Cephalosporin Antibiotics, Portschr. Chem. Org. Naturst, 28, 343-403 (1970), in particular on pages 379-393; the now famous "Woodward Intermediate" is going on there Shown on page 387 as compound 146.

509820/1161

^{Μ / 12β60} :, 23S68G2

There have been releases within the last few months. appeared describing "new works" and older works summarize and quote:

a) From Imperial College, London, and Glaxo together with DHRBarton et al _o , J. Ghern. Soc. (C), 1971,

b) from Oxford University by D.M. Brunwin et al., J. Chem ,. Soc. (C), 1971, 3756-3762;

c) from the University of Newcastle upon Tyne, by B.G. Ramsay and R.jr.Stoodley, J. Chem. Soc. (C), 1971, 3859-3867;

d) from Lilly through S. Kukolqa, J. Amer. Chem. Soc. 93 » 6267-6270 (1971); .

e) by Lilly by G.E. Gutowski, et al., Tetrahedron Letters No. 37: 3433-3436 (1971);

f) by Lilly a series of publications called Chemistry of Cephalosporin Antibiotics, for example No. 25 in J. Medicinal Chemistry 14 (1 ) , 11-3.6-1138 (1971) and No. 21 in J. Qrg «Chem. > 36 (9) . 1259- ^ 1267 (1971). '■' · ■

The anhydride compounds used as starting materials in the present invention were first described by Saul Wolfe in J. Amer. Chem. Soc. 85,, 643-644 / March 1963), the Belgian. . See patent specification 621 452 and iJS-rES 3 311 638 described. Further subscribers to anhydropenicillins through Saul Wx> lfe include Can. J. € hem. 46, 459; and 2549 (1968). ''

Various azetidinones have been described in patents of fenbart, so "as described in US Patents 3,487,070, 3,487 W ^1" 3487 £ 72 (Class 260-239) and 3,487,090 (lOLass; e 2 ^ 0 ^ 326) of Sheeha ^ iwid, for example Woodward ^r s W-2S- 3 483 215 (class 260-306 ^ 7) and US-PS 3 449 336 _f C class 26Ö-243) \ ^{ -

- _: 3, - ■

Μ / 128βΟ

The oxazoline of the structure

CH-CH

C - OC

Il

H,

which is commonly described as

CO ₀ G

2 ^GEE 3

has been prepared from the methyl ester of benzylpenicillin in 18 yield as described by Barton et al., J. Amer. Chem. Soc., 91, 1529 (1969) and a), supra. Just Another bicyclic, sulfur-free compound of this type has been described [by F.G.Brain et al.,

- k - 509820/1161

M / 12860

62

J. Chem. Soc. Chem. ' Comm., 229-230 (1972) 3, However, one is expedient synthesis of such esters or acids not published. \ /

The stereochemistry of the methyl ester of benzylpenicillin (i.e. the penicillin G) produced by fermentation is like is shown as follows:

Kukolja [J. Amer. Chem. Soc. 93, 6267-6270 (1971)] chlorinated (with two equivalents of chlorine) methyl 6-phthalimidopenicillanate

HH

> —N- 0 (f »

GH

being a mixture of the two epimers of the structure

originated.

CO ₂ CH ₃

509 8 2071161

Outline of the invention

The invention creates a number of processes (and certain new intermediate products produced as a result, which begin with the chlorination of an anhydropenicillin. ;

The processes are listed schematically in the usual manner directly below, with the starting material being exemplified Phthalimido-anhydropenicillin of structure

is needed. For the sake of simplicity, this link is written below in the following form:

TFA means trifluoroacetic acid;
DIC means diisopropylcarbodiimide;
HBS means N-bromosuccinimide;
tert-BuOH means tert-butyl alcohol; Ph means phenyl,

FT means phthalimides

509820/1161

The invention includes the reaction schemes set out below a, which only explain the invention, however, they do not limit:

5099167-1.181

M / 12860

H H Ftp »-,

-N

3a

.N

'COCl

4a

CH ^ OH

Cl H Cl

H Cl

-N.

Ptfe »

COCl

(Cf. Kukolja, supra) * *

Pt 6 »

tert-BuOH H Cl

C0 ₂ t-Bu

TPA

Pt «» ·

H_ Cl

LJ

■ COOH

4c

H Cl

FtP "COCl

4b

8a

Br Br

. 5,09820 / 116.1;

M / 12860

H Cl

356862 '

Ftp

CO ₂ Me. ■ '·

-9- - ■; \; - 5Q982JD / T161,

Μ / 128βΟ

«· ■

• A

O - C - H

H Cl

H Cl

Ft »

8 2 0/1161

M / 12860

The above and additional procedures and intermediates according to the present invention are listed below, the starting material being the toluenesulfonic acid salt (TsOH) is used by 6-aminoanhydropenicillin.

This connection has the structure:

TsOH-H ₂ Ni

I IA

C C = O

which are shown below for simplicity as follows
will:

Ts *

■ *

- 11 '■ - ·

5098 20/1 161

M / 12860 Ts *

H Cl

■ Ν

H Cl

; 1. NaHCO-

2.

(one isomer isolated) ■ Η Cl

■ Ν

H Cl

Ft * -
O ^

H Cl

CO ₂ CH,

6b,

-12- '

5 0.9 8 2 07 1161 ;:

M / 12860

H . Cl

Ts

- ο

'■

5c

COOH - ■

NaHCO ₃

H H

Nl /

CO ₂ H PhCH ₂ COCl ' H Cl

.N,

COOH

NaHCO.

• \ /

H Cl

CO ₂ H

■ \ PhCH ₂ COCl

HH

PhCH ₂ CONH-

COOH PhCH ₂ CONH

- 13 '- v ..

ORIGINAL INSPECTED

M / 12860

DIC

H Cl (jiCHgCONH * r -T

-N

K Cl

H ₂ N

CO ₂ CH ₅

Silicates ¹

CH ₂ Ph

Kiiii

CO ₂ CH ₅

L-N,

CO ₂ OH,

6a

H Bi

3O ₂ CH ₅

DIC

"3"

^ CH ₅ COHH -

6d

Co ₂ CH ₅

^: 56 9 8.2 0./1 161V ,.

ORIGINAL INSPECTED

η. approx

-N.

^ CH ₀ COOH

DIG

(J) CH ₀ CONiI r

spontaneous ' - ■

Cyclization '

CH ₀ EtW "·

Huri

.NSECO-

, earner

le

CO ₂ CH ₅ .2

O

(J) CH ₂ COOH

Slide. .

• 7c

: 2NBS
■ "V

"= 5 ■

.Br

Br

-15-

·: ORSß ^ ÄO INSPECTED

M / 12860

H Cl

H Cl

Ft

ke

COpCH,

2 parts

ο. '

CO ₂ CH

■ 2 ·

Et ,, NCl

H Cl

FTP

CO

3-5 parts

CO ₂ CH,

H Cl

-m;

H Cl ".

1 part -16-

H Q.1

CO ₀ CH,

²⁵ 6b

β parts

50.9820 / 11 6Ί

ORIGINAL INSPECTED

/ 2350862;

M / 12860 * n

Thus, direct chlorination (excess Cl ₂ , CH _{2 Cl 2} , 20 ° C, 3 min.) Of anhydro-e-phthalimidopenicillin (3a) followed by removal of the solvent under reduced pressure yields a 3: 2 mixture of two isomeric compounds. The major isomers 4a of melting point 210 ⁰ C with decomposition is obtained in 38 ^ yield (from 3a) by crystallization from CHCl ^ -Petroläther (calculated for C ₁₆ H ₁₂ N ₂ O ₄ Cl: C 52.3; H 3.31 ; N 7.69; Cl 19.4. Found: C 52.03; H 2.96; N 7.76; Cl 19.53),

²⁴² ( ^Λο

The infrared spectrum of 4a (and of the mixture of 4a and 4b) shows peaks at 5.48 (COCl), 5.55 (ß-lactam), 5.62, 5.79yu (phthalimido).

The hydrolysis of 4a (boric acid borax buffer [* Saul Wölfe, R. N. Bas sett, S. M. Caldwell and F.T .. Wasson, J. Amer. Chem. ·. Soc., 91, 7205 (1969)] or Aqueous Acetone) provides the Acid 4c from melting point 168 to 174 ° with decomposition in 73 l & lger yield. '

(Calculated for C ₁₆ H ₁₃ N ₂ O ₅ Cl: C 55.05; H 3.76; N 8.02; Cl 10.02. Found: "'C 54.65;" H 3.79; N. 8.46; Cl 10.55)

f 236 (7f00), 296 (1,800); ;

. 5.52, 5.60, - 5.80, ⁴ '5.85 / U ..

Methylation of 4c (CH ₂ N ₂ , At ₂ O-CH ₂ Cl ₂ ) gives the ester

4e of melting point 178-180 ⁰ C'unter decomposition in 97% yield. The same compound is obtained from ^ a on treatment with absolute methanol (30 min., 20 ° C) in 93% yield. The treatment of the mixture of 4a and 4b with methanol (30 min., 20 C ⁰⁾ and crystallization of the resulting residue (CHCl ^ -Petroläther) provides 4e in 55% yield · chromatography of the remaining parent

509820/1181

Caustic solution (Woelm aluminum oxide, activity level II; 1: 1 CH ₂ Cl ₂ : C ₆ H ₆ ) leads to 22 % of 4f, the methyl ester derivative of 4b with a melting point of 192 to 194 ° C in the first fractions, followed by an additional 9% 4e. The tert-butyl ester 4g is obtained in 33 % yield by refluxing 4a in anhydrous tert-BuOH for 21 hours. Dissolving 4g in trifluoroacetic acid and evaporating the solvent after 3 minutes gives 4a in quantitative yield.

The relative amounts of 4a / and 4b are not significantly affected by chlorination in the presence of excess Et ^ NCl; other chlorinating agents (SOpCl ₂ , pyridinium trichloride, pyrrolidone hydrotrichloride) give more complicated mixtures. The esters 4e and 4f are equilibrated with excess Ät.NCl [under identical experimental conditions the same mixture is obtained from both directions], the trans isomer 4f predominating. Depending on the conditions, ratios varying from 3: 2 to 5: 2 are obtained and the isomers can be separated by chromatography as described above.

The reaction of sodium azide with 4e [4c decomposes under these conditions] (DMF 90 °, 3 hr.) Gives the trans-Azid'4h with melting point 144-145 ⁰ C (67%); Under the same conditions, 4f is converted to 4i with a melting point of 183-187 ° C (52 %) . The reaction of 4f with Et ^ NOAc "(pure CHCl", reflux, 17 hrs.) Gives the cis-acetate 4j (62 %). The reaction of 4f with tetramethylguanidinium formate [an agent invented for this purpose] leads to the cis-formate 4k (35 %).

The above reaction sequence was carried out with various modifications to the para-toluenesulfonic acid salt [S. Wolfe, Can. J. Chem., 46, 459 (1968)] from anhydro-6-aminopenicillin (3b). The chlorination (0 to 5 ° C, CH ₂ Cl ₂ ) followed by

- 18 509820/1161

M / 12860

The removal of the solvent and trituration with ether gives a v ^ i-MischurigvoiL ^ ₁ and gb in the form of a stable (below,., 20 ⁰ C) white powder with a melting point of 148 to 149 ⁰ C with decomposition, the hydrolysis of this powder ( Acetone / water) _; results in an Ari mix. of acids 5c and 5d. Alternatively, treating the mixture of 5a and 5b provides _; Methanol and crystallization from chloroform / hexane give the pure cis-methyl ester from melting point 125 to 130 ° C with decomposition in 67% yield. The neutralization of ge (NaHCO ₅ ) - and reaction of the free base 6a with JK-carbethoxyphthalimide [GHL Nefkens, Nature, 185 , 309 (1960)] gives 4e, - whereby it is confirmed that the chlorination reaction is in the same catfish , with the two anhydro- ^ penicillins. . "_. '." -.......:. · "

The equilibration of 6a with the trans isomer en 6b proceeds smoothly in the presence of chloride ions, the best ratio being achieved in favor of 6b (6: 1 ) with tetramethylguanidinium chloride (CH ₂ Cl ₂ ^ 12 h reflux). The isomers are separated by aluminum oyster chromatography. The cis-6a Aminochlorid reacts smoothly with tetramethylguanidinium azide (2 equivalents ^ ^ _CHCl; 1 h »reflux) (Ä.J. Bapa, J.Org.Chem, 32, 1426 (I966).), The trans -Aminoazid 6d (90 %) is obtained; the cis amino azide 6c with a melting point of 116 to 11: 7 ° C is obtained in the same quiet manner from 6b in 85% yield,.

The 5-chloropenicillin G analogue £ §. yom melting point 111 to 115 ⁰ C (decomposition) is obtained in 82 9oiger yield in the Phenylacetylierung of 6a (PhCH ₂ COOH, diisopropylcarbodiimide (DIC), CHpCl _0). The C5 epimer 7b, which is obtained in the same way from 6b, cyclizes spontaneously to give the oxazoline 2 ^ [JC Sheehan, "Molecular Modification in Drug Design", Advances in Chemistry Series, No. 45 _f American Chemical Society, Washington, DC ₅ 1964, page 15; D »HR Barton,

. ; - 19 - ■ '■ "" ■■■■ ■ - ■ -. ■

509820/1161

M / 12860

F. Comer and P.G. Sammes, J. Amer. Chem. Soc, 91, 1529 j

(1969)] from melting point 126.5 to 127 ° C; additional amounts of 2 are obtained from 7a by rapid chromatography on silica gel or aluminum oxide [if the trans-acylaminochloride 7b is shaken with bicarbonate, varying amounts (up to 10 %) of 7e can be isolated in addition to the oxazoline]. The 5-azidopenicillin-Gr analogue 7c from! Melting point 102 to 103 ° C. is obtained in 83% yield in the phenylacetylation of 6c (PhCH ₂ COOH, diisopropylcarbodiimide, CH ₂ Cl ₂ ); the C5 epimer 7d is in. obtained in the same way from 6d or by reaction of 7a with tetramethylguanidinium azide (CHCl ,, reflux). *

The allyl oxidation of 4e with N-bromosuccinimide (NBS, 2 equivalents, COIr ₉ benzoyl peroxide, Photoflood No.2 lamp [S. Wolfe and DVC Awang, Can, J. Chem., 49, 1384 (1971); this result should correspond to the behavior of the underlying anhydropenicillin versus allyl oxidation, S. Wolfe, C. Ferrari and WS Lee, Tetrahedron Letters, 3385 (1969). As noted, S. Wolfe, RN Bassett, SM Caldwell and FI Wasson, J. Amer. Chem. Soc ,, 91_, 7205 (1969), the enamine character of the double bond of these compounds is not found in the monocyclic systems]), is complete within 5 minutes. The product 8a crystallizes from CCl ^ petroleum ether in 81 % yield and has a melting point of 70 to 72 ° C. with decomposition. Under the same conditions, the C5 epimer 4f gives 8b in 70% yield. The extraordinary ease of this NBS oxidation is found under the same conditions in the successful functionalization of 7c and Z ^ . The methyl groups of 4e and 4f show no significant difference in reactivity. Oxidation of 4e with one equivalent of NBS yields a 1: 1 mixture of 8c and 8d. Treatment of this mixture with tetramethylguanidinium azide (1 equivalent, CHCl ,, 3 hours, 20 ° C [the 5-chloro-substituent is stable under these conditions])

- 20 509820/1161

M / 12860/4

provides a 1: 1 mixture of 8e and 8f :;

4.7 (AzId), 5.55 (β-lactam), 5.62, 5.7.9 (phthalimido), 5.78 (ester). Hydrogenation (PtO ₂ , benzene, 3.16 kg / cm (45 psi) 6 hours) provides the mixture of amines 8g and 8h; XmaS " ³ⁱ⁵ ' ⁵⁵ ' ⁵ ' ⁶² ' ⁵ ' ⁷⁹ ' ⁵ » ⁸³ / ^u · This mixture is recovered unchanged after refluxing in chloroform. Treatment with K-tert-BuO-tert-BuOH (20 ° C, 1 hour), followed by careful chromatography, (neutral aluminum oxide, activity level II, eluting with graded mixtures of CgHg-CH ₂ Cl ₂ ) gives a crystalline one in 12% yield (24 % if only one reacts in 8g or 8h) Compound with a melting point of-121 to 122 ⁰ C, which is based on its infrared spectrum (Xmax ^ »92» 5.56, 5.62, 5.79, 5.82, 6.03 / u) and the nuclear magnetic resonance spectra (2nd , 25 (4H), 3.87 (1Ή, d, 2.0 Hz), 4.30 (1H, d, 2.0 Hz), 6.23 (3H), 7.70 "(2H), ' 7-88 (3H)), which has structure 9. '

Another important embodiment of the present invention is schematically explained in the following, where R has the meaning of N-propyl or tert-butyl and HMPT

0
(CH ₃ ) ₂ NPN (CH ₃ ) ₂

means:

. - 21 -

5 09820/1161

M / 12860

CH ₂ Ph

N · 0 Hii "| / muH

COOH

JT

Hm

iiH

CO ₂ CH ₅

1. RSLi / HMPT, 2. H ^ O ⁺

H-

H H.

(as in the US patent
5,499,909)

PhCH ₂ CONH ■ ·

'' COOH

3. CHjOH

- 22 -

⁴ 3

CH.

COOH '

50.9 8 20/1 161

M / 12860

¥ f s / θ 0 1. Cl ₂ ■ 2. H ₃ O ⁺ 3.
4th H ₂ O ;;

^{H H}

Tt ι

0 '

^J J »« »'uC00H

3a

fs

or.CH ₃ NH ₂

(■ as in S. ¥ olfe f - and SK Hasan ^ Can * J. Chem., 48, 3572 (1970)); ">

- 23 -

5038 20/1161

M / 12860

H Cl

H Cl

H Cl

Ftp

-N,

CO ₂ CH ₅

1 NBS

Br

4- H Cl

CO ₂ CH ₃

H Cl

FTP

CO ₂ CH ₅

Br

5 0 9 8 2 0/1 I 6 1

Μ / 128βΟ

The above and in the examples are expressed more generally described method is carried out with numerous side chains at the 6-position, "for example by the starting reagent is an anhydropenicillin of the formula

R - CH - CH

N-

C = C (CH ₃ ) ₂

■ where R is acylamino., Including phthalimido or amino is used.

Thus, in the methods disclosed herein, instead of the Phthalimido group (Ft) or the free amino group or the tosylate (Ts) of the corresponding reagents, also intermediates and products used in which each of these groups

1 pen by an acylamino group R of the formulas

CHC

R ^J O

I 11

NH-; Ar - Χ "- C-C - NH

0
Ar - C- NH -

OR '

- NH -

Ar

- 25 -

- NH -

509820/1161

M / 12860

Il

■ ο

Il

C-NH R ⁹

JtS

δ -

- NH -

Ar-NH-C -NH -

OH NH

ζ ¹ ο

I Il

C-C

-NH-

and

I!

Il

R ¹¹ , - NH - C - NH - and 'R ¹² - O - C - NH -

is replaced,., wherein R is a group selected from hydrogen, Amine, carbobenzoxyamino, phenyl, fluorine, chlorine, Means bromine, iodine, hydroxy, lower alkanoylpxy and lower alkoxy;

X is a group selected from oxygen and sulfur;
R ^ and R each represent a group selected from hydrogen, phenyl, benzyl, phenethyl and lower alkyl;

7th
R 'is lower alkyl; 8 9

R and R are each a group selected from lower alkyl, Lower alkylthio, benzylthio, cyclohexyl, cyclopentyl, cycloheptyl, benzyl, phenethyl, phenylpropyl, furyl, thienyl, Naphthyl and Ar, ~ mean;

- 26 -

509820/1161

Μ / 128βΟ

Ί Π
R - a group selected from lower alkylamino, di (lower) alkylamino, cycloalkylamino having 3 "to 7 carbon atoms including, allylamine, diallylamino, phenyl (lower) alkylamino, morpholino, lower (alkyl) morpholino, di - (Lower) -alkylmorpholino, morpholino- (lower) -alkylamino, pyrrolidino, lower alkylpyrrolidino, di- (lower) -alkylpyrrolidino, Ν, Ν-hexamethyleneimino, piperidino, lower alkylpiperidino, di- (lower) -alkylpiperidino, 1,2,5 , 6-Tetrahydropyridino, N- (lower) -alkylpiperazino, N-phenylpiperazino, N- (lower) -alkyl-lower-alkylpiperazino, N- (lower) -alkyl-di- (lower) -alkylpiperazino, furfurylamino, tetrahydrofurfurylamino, N-lower alkyl N-furfurylamino, N-alkyl-N-anilino and

Lower alkoxyanilino means. ·.

12 3
Z, Z and Z each represent a group selected from lower alkyl and aryl;

11 ·

R is a group selected from lower alkyl, lower cycloalkyl, Naphthyl, benzyl, phenethyl and

I! ·

Ar-C- means;

12 '"" "■" ■

R 2,2,2-trichloroethyl or benzyl and Ar, 'a monovalent Remainder with one of the formulas »

R ¹ '■ R ¹ R1

Il> l

■ r ²

12 3

in which R, R and R ^ each represent a group selected from hydrogen, chlorine, "bromine, iodine, trifluoromethyl, phenyl, lower alkyl and lower alkoxy, but only one of these radicals R, R and R ⁵ can be phenyl .

- 27 5098 207 1 16 1

M / 12860

For example, a preferred group becomes more inventive Intermediate and end products on the basis of end products made by acids, explained with the following formulas:

O NH - CH- Q -CH ir I. I. CH - C - O - c - -N τ
R.

C (CH ₃ ) ₂ CHCOOH

wherein R is lower alkyl;

It

-C-NH-. CH-CH

CH-COOH

1 2 3

wherein R is lower alkyl and R and R are each a group under hydrogen and chlorine, - mean,

Il

• 0

C-NH-CH-CH

. I * I

0 = »C ^ N-

CH-COOH

1 2

wherein R is lower alkyl and R is a group selected from

Mean hydrogen and chlorine;

- 28 -.

509820/1161

M / 12860

O - NHR ' CH- > ° \ Il I. C. -NH- O
1! c- O = CH C (CH-,) ,, I. N CH-COOH

wherein R is lower alkyl;

CH - C - NH - CH 0 - C - R

I! 0 Il

CH C (CH ₃ ) ₂

CH-COOH

wherein R is lower alkyl;

OR.,

_ ^

C-NH-CH-CH \ M- I

O = C-N--

wherein R is lower alkyl; .CH-COOH

f R. \\ - o - CH
= / ■■; ■ R 0 CH
I.
N
"■.> S CH ₅
I ²
C - CH ₃ { - C - NH COOH- Lower alkyl - - wherein means
mm- - CH-
I.
O = C—- • -
>
29 -

509820/1161

M / 12860

, 3 / V = I.

"N.

H • Ν

NH - OH - CH

I.

O = C N

I.

CH ₂

C -

COOH

1 2 5

wherein R is lower alkyl and R and R are each a group

are selected from hydrogen and chlorine;

1 2

wherein R is lower alkyl and R is a group selected from water

mean hydrogen and chlorine;

- NHR

-NH-CH-CH-CH

I · I

O = C N

CH ₂ C -

wherein R is lower alkyl;

509820/1161

M / 12860

11

CH- C-NH- CH- CH "

-. C- R

11

CH ₂

O = G.

• C ·
COOH

wherein R is lower alkyl; .,

OR

■ H

-C «

NH - CH-

■ I

O = C-

CH

-N

C - ".CH ₃

C ^: COOH

■ where R means lower alkyl.

The present invention creates the process for manufacturing an antibacterial agent, which thereby characterized is that one is a compound of the formula

H ₂ N -

• 0 «

CH -

It

CH C (CH ₃ ) 2 N -: CH - COOH

or a salt thereof with an organic monocarboxylic acid chloride or a functional equivalent thereof, and also the process for producing one antibacterial agent, which is characterized in that one is a compound of the formula

- 31 -

509820/1161

H
• Ν

H ₀ N - CH-CH

ι · ι

O = C N C -

COOH

or a salt thereof reacts with an organic monocarboxylic acid chloride or a functional equivalent thereof leaves. '·

In a preferred embodiment, the present invention provides the method for producing an antibacterial agent which is characterized in that one connection of the formulas

CH-CH -C (CH,) _O HpN-CH CH CH ₀

Il I ^{3 2} or ^ III ²

C - N- OH-COOH O = C N 'C-CH-

COOH

or a salt thereof with either an organic monocarboxylic acid chloride of the formulas

0 R ⁵ 0 0

Ar-CHC-Cl; Ar-X-C-C-Cl; Ar-C-Cl;

509820/1161

M / 12860

Ό'-

- Cl

II

■ C - Cl. ; -.R ⁹

Ar

Il

It

C -. Cl

.- R

- Cl

X Ar-NH-C-Cl

Z ² -

"1

^z 0

t It

C-C

-Cl

^ \ 0
It
ft .- PH X - Cl L. f - Cl ^^ Il
0 and R ¹¹ X
Il
- NH -> C

wherein ^ R is a group selected from hydrogen, amino, Carbobenzoxyamino, phenyl, fluorine, chlorine, bromine, iodine, hydroxy, Means lower alkanoyloxy and lower alkoxy; X is a group selected from oxygen and sulfur;

R and R are each a group selected from hydrogen, Denotes phenyl, benzyl, phenethyl and lower alkyl; R ^ is lower alkyl; :.

R and R ^ each represent a group selected from lower alkyl, Lower alkylthio, benzylthio, cyclohexyl, cyclopentyl, cyclo-

Β09 8 20 / 1Ί61

heptyl, benzyl, phenethyl, phenylpropyl, furyl, thienyl,

Naphthyl and Ar '"mean;

R is a group selected from lower alkylamino, di (lower) alkylamino, Cycloalkylamino with 3 to 7 carbon atoms including, Allylamino, DialIyIamino, Phenyl- (lower) -alkylamino, morpholino, (lower) -alkylmorpholino, Di- (lower) -alkylmorpholino, morpholino- (lower) -alkylamino, Pyrrolidino, lower alkyl pyrrolidino, di- (lower) alkyl pyrrolidino, N, N-hexamethyleneimino, Biperidino, (lower) -alkylpiperidino, di- (lower) -alkylpiperidino, 1,2,5,6-tetrahydropyridines, N- (lower) -alkylpiperazino, N-phenylpiperazino, N- (lower) -alkyl- (lower) -alkylpiperazino, N- (lower) -alkyl-di- (lower) -alkylpiperazino, Furfurylamino, tetrahydrofurfurylamino, N- (lower) -alkyl- N- furfurylamino, N-alkyl-N-anilino and (lower) -

means alkoxyanilino;

12 3
Z, Z and Z each have a group selected from low-

alkyl and Ar 'are;

11 ''

R is a group selected from lower alkyl, lower cycloalkyl, naphthyl, benzyl, phenethyl and

Ar-C- means; and Ar- is a monovalent residue of one of the Formulas

and R ² -U-

12 ^

means in which R, R and R are each a group selected under hydrogen, chlorine, bromine, iodine, trifluoromethyl, Phenyl, lower alkyl and lower alkoxy mean, but only

- 34 -

509820/1161

a group R can be phenyl; or with a functional equivalent of the acid chloride

lets react. . "

The term "lower ■ rigalkyl" used in the present invention means both straight-chain and branched "al I-phatic hydrocarbon radicals with 1 to 10 carbon atoms, like methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, amyl, hexyl, 2-ethylhexyl, heptyl, decyl etc. In the same way, the term; "Low" in those cases where it is used as part of the description of another group, for example "lower alkoxy" is, on / the alkyl portion of such a group and has hence the same meaning as above in connection with "Lower alkyl" was described »"

The functional equivalents of the above acid chlorides used as acylating agents for a primary amino group include the corresponding carboxylic acid bromides, acid anhydrides, including mixed anhydrides, and especially those of stronger acids; such as the lower aliphatic monoesters of carbonic acid, of alkyl and aryl sulfonic acids, and mixed anhydrides produced by more sterically hindered acids such as diphenylacetic acid. Furthermore, an acid azide or an active ester or thioester (for example with p-nitrophenol, 2,4-dinitrophenol, thiophenol, thioacetic acid) can be used, or the free acid can be coupled with the primary amine itself, after first this free acid with Ν , Ν'-dimethylchloroforminium chloride [GB-PS 1 008 1? 0 and Novak and Weichet, Experientia XXI / 6, 360 (1965)], or through the use of enzymes or an N, N ^T -carbonyldiimidazole or, a Ν , Ν'-carbonylditriazole [South African patent 6: 5/2684], or a carbodiimide

- 35 -

50 9 82 0/1161

Reagent [esp. NjN'-dicyclohexylcarbodiimide, Ν, Ν'-diisopropylcarbodiimide or N-cyclohexyl-N ¹ - (2 ^ morpholinoethyl) -carbodiimide; See Sheehan and Hess, J. Amer. Chem., Soc. 77 , IO67 (1955)] or an alkynylamine reagent [R ,; Buijle and HG Viehe, Angew. Chem. International Edition J5, 582 (1964)] or a ketenimine reagent [CLStevens. And MEMonk, J. Amer. Chem. Soc. 80, 4065 0958)] or an isoxazolium salt reagent [RB Woodward, RAOlofson and H. Mayer, J. Amer. Chem. Soc. 83, 1Q10 (1961)]. Another equivalent of the acid chloride is a corresponding azolide, ie an amide of the corresponding acid whose amide nitrogen is a member of a quasi-aromatic 5-membered ring which contains at least two nitrogen atoms, ie imidazo !, pyrazole, the triazoles, benzimidazole, benzotriazoles and their substituted ones Derivatives. As an example of the general method for preparing an azolide, Ν, Ν'-carbonyldiimidazole is reacted with a carboxylic acid in equimolar proportions at room temperature in tetrahydrofuran, chloroform, dimethylformamide or a similar "inert solvent, with the release of carbon dioxide and one mole of imidazo }, the carboxylic acid imidazolide is formed in virtually quantitative yield. The by-product imidazole precipitates, and can be separated and the imidazolide can be isolated, but this is not essential. The methods of carrying out these reactions to produce a penicillin and those of isolating the so The methods used to produce penicillins are well known in the art, and dicarboxylic acids give diimidazolides.

The nontoxic, pharmaceutically acceptable salts include metal salts such as the sodium salt, the potassium salt, the calcium and aluminum salts, the ammonium salt and substituted ammonium salts, for example salts of nontoxic amines such as trialkylamines including triethylamine, procaine, dibenzylamine, N -Benzyl-ß-phenethylamine, 1-ephenamine, N, N ¹ -dibenzylethylenediamine, dehydro-

- 36 509820 / ΐ161

abiethylamine, NjN'-bis-dehydroabietylethylene-diamine, N- (lower) -alkylpiperidines, for example N-ethylpiperidine ■ and other amines which form salts with benzylpenicillin be used· · .

In particular included within the scope of the present invention are the antibacterial agents which are used in the acylation process described above by the local use of the organic monocarboxylic acids or their acid chlorides or other equivalents which were previously used for the acylation of 6-aminopenicillanic acid, as for example in US-Psen 2 941 995, 2 951 839, 2 985 648, 2 996 501, 3 007 920, 3 025 290, 3 028 379, 3 035 047, 3 040 032, 3 040 033, 3 041 332, 3 041 333, 3 043 831 , 3 053 831, 3 071 575, 3 071 576, 3 079 305, 3 079 306, 3 080 356, 3 082 204, 3 093 547, 3 093 633, 3 116 -28S, 3 117 119, 3 HS 877, 3 120 512, 3 120 513, 3 120 514, 3 127 394, 3 140 282, 3 142 673 »3 147 247, 3 174 964, 3 180 863, 3 198 804, 3 202-653, 3 202 654, 3 202 655, 3 210 337, 3 157 639, 3 134 767, 3 132 136. And in the GB ^ PSs 874 414, 874 416, 876 516, -876 662, 877 12.0, 877 323, 877 531, 878 233, 880 042, 880 400, 882 335, 888 110, 888 552, 889 066, 889 069, 889 070, 889 168, 889 231, 89 0 201, 891 ^ 74, 891 279, 891 586, "891 777, 891 938, 893 518, 894 247, 894 457, 894 460, 896 072, 899 199, 900 666, 902 703, 903 785, 904 576, 905 778, 906 383, 908 787, 914 419, 916 097, 916 204, 916 205, 916 488, 918 169, 920176, 920177, 920 300, 921 513, 922 278, 924 037, 925 281, 931 567, 932 644, 938 066, 938 321, 939 708, 940 488, 943 608, 944 417; in numerous Belgian patents, for example 593 ²²² »595 171, 597 859, 602 494, 603 703, 609 039, 616 419 and 617 187 and in South African patents, for example 60/3057, 60/3748, 61/1649, R61 / 2751, "62/54, 62/4920, 63/1612 and 63/2423.

• - 37 509820/1161 ,

«Yeah

¥ enn the acylamino group of the compounds according to the invention also contains a strongly basic group, for example a primary amino group, as is the case in the preferred Embodiments of the formulas

Ar - CH - C - NH - CH - CH

t 11 \ ^J

^m 2 O = CN CHCOOH

and

H 0 ^ <

Ar - CH - C - NH - CH - CH CH ₅ -

O = CNC - CH _x

COOH

in which Ar has the same meaning as above (and R, R and R in Ar are preferably hydrogen), the products are amphoteric and are normally in the zwitterion form before, but can form acid addition salts, such as for example with non-toxic, pharmaceutically acceptable organic Acids such as acetic acid, citric acid, succinic acid, ascorbic acid and the like and with inorganic ones Acids such as hydrochloric acid, hydrobromic acid, Sulfuric acid, phosphoric acid and the like.

The methods of the invention also include the use of the compound of the formula

- 38 -

50982071161

JS

H ₀ N-CH-CH

■ ί- Γ '

O = C-N-CHCOOH

and their salts with acids and bases and the compound the formula

H-.

- CH.- CH CH ₂

C · N C - CH,

0 .- ■ - ^ C ^

COOH ■

and their salts with acids and bases as "final reagents"^; . ·. .

These "end reagents" are produced, for example, by making the compound of the formula

C ₆ H ₅ CH ₂ O - C - NH - CH - CIt

C - T - N - CH.

Il I

0 O = C

subjecting to hydrogenolysis, or by removing the compound of formula

- 39 - ■

509820/1161

M / 12860

0
it

■ N

-NH- CH - CH

CH

I.

Il

.N

^ c

C - CH,

subjecting to hydrogenolysis, or by either breaking the compound the formula

(CH ₃ ) ₃ C _; -

It

C -

- NH - CH - CH II

C N

it "v

H • N

"-CH ₂ O = C-O-C (CH _x ).

or the compound of the formula

(C ₆ H ₅ ) ₃ C - NH - CH-

ti

-CH I

CH ₂

> C ι

O = C-O-

subjecting to an acidic cleavage, or by breaking the compound the formula

- 40-

509820/1161

Cl ₃ C-CH ₂ QC-NH-CH-GH C (CH ₅ )

C-N-CH

»

⁰ O = C

OCH ₂ CCl ₃

or the compound of the formula

• ο ■ I
I. ι -
O = C - C - CH,
> ■ -.- .. · -3. Cl ₃ C-CH ₂ OC-NH - N OCH ₂ CCl ₃ o < -

treated with zinc and acetic acid.

Such esters are produced, for example, by using an anhydropenicillin of the formula as starting material

R ¹ - CH-CH C = O

0 '

I I i

.C_-NC = C (CH,) ₂

in the R

- 41 -

5098 20/1161

ο ο

C ₆ H ₅ CH ₂ OC-NH-, (CH ₃ ), COC-NH-,

Cl.O
Cl-C-CH ₂ OC-NH-,
Cl

Is formyl, o-nitrophenylsulfenylamino or o-nitrophenoxyacetamido, or its equivalents as described in US Pat. No. 3,271,409; used and in the appropriate later stage (the reaction with acid chloride) the methanol either against tert '. -Butyl alcohol or trityl alcohol or trichloroethyl alcohol replaced; or 'in the later stage (der.Reaktion with carboxylic acid) the diazomethane is replaced by either diphenyldiazomethane, 2,2,2-trichloroethyl chloroformate, benzyl chloroformate or tert-butyl chloroformate.

These esters are then converted into "final reagents" by the ester is removed and the blocked amino group either sequentially or simultaneously into a free amino group convicted. So if the blocked amino group is phthalimido, it becomes by treatment with hydrazine hydrate in dioxane be! "about room temperature for at least 12 hours or with methylamine in aqueous dioxane in

« Ti

converted to a primary amine. When the blocked amino group means carbobenzyloxyamino, "it becomes catalytic Hydrogenation, converted to a primary amino group, leaving the product behind. When the blocked amino group Formyl, carbo-tert-butyloxy-amino or o-nitrophenylsulfenylamino means it is either by reacting with anhydrous hydrogen chloride in a non-protonating one Solvent such as benzene or methylene chloride or by reacting with trifluoroacetic acid into a primary Amino group converted. The · Trichloräthoxycarbonylaminogruppe is made by reaction with zinc in aqueous acetic acid converted into a primary amino group. In addition,

- 42 509820/1161

Μ / 12860. ; YS

■ if the "blocked amino group is carbo-tert-butyloxyamino is the final two reactions by using a stronger acid for longer periods of time and optionally carried out at higher temperatures in one stage. . :

When the blocked amino group is o-nitrophenoxyacetamido means it is converted to a primary amino group either

Ca) by catalytic hydrogenation (for example in water at room temperature using 30 % palladium on diatomaceous earth). and then allowing the mixture at an acidic pH (for example in water with 20! & Lger hydrochloric acid was acidified to a pH of 2, at about 10 ⁰ C for at least 20 min., or at 25 ° C for at least 24 hrs.) or

(b) by rapid, for example over 1 to 3 minutesj, adding the blocked compound (for example 7 mmol) in cold water, for example 30 ml, to 5 % Pd-C (for example 0.05 g) _; which is suspended in a cold solution of KBH ^ (for example 14 mmol), dissolved in water, for example 70 ml,

convicted. · "''.

- 43 -

509820 / 116t

Μ / 12860

Chlorination of anhydro-6-phthalimidopenicillin

Chlorine gas is passed into a solution of 500 mg (1.52 mmol) of anhydro-6-phthalimidopenicillin in 15 ml of methylene chloride at room temperature. The introduction of the gas is ended after 3 minutes and after a further 5 minutes the yellow solution is evaporated to dryness. The resulting white foam was crystallized from a mixture of chloroform and petroleum ether to give 210 ⁰ C are obtained by decomposition in two portions a total of 250 mg (32%) of melting point material. The infrared spectrum of the remaining mother liquor is identical to that of the crystalline material listed below.

Analysis C ₁₆ H ₁₂ N ₂ O ^ Cl ₂ :. ·

C H N Cl

calc .: 52.3 3.31 7/69 19.4 %

found: 52.03 2.96 7.76 19.53%

The infrared spectrum shows peaks in, the carbonyl region at 5.48, 5.55, 5.62 x and 5.79 / u.

The NMR spectrum shows peaks at 2.13 (4H), 3.78 (1H, d, J = 4.2 Hz), 4.24 (1H, d, J = 4.2 Hz), 7.60 (3H), 7.69 (3H). "

The UV spectrum shows a ^ ^ J2 ^H of 242 (10,000).

The connection has the structure

- 44 -

509820/1161

Hydrolysis of the dichloride

500 mg of the dichloride (1.36 mmol) in 500 ml of dimethyl sulfoxide is added ^{dropwise at 0} ° C. with 50. "Ml of a buffer - of pH 7.95 (prepared by diluting 50 ml of 0.1 molar boric acid and 2 , 6 ml 0.1 molar NaOH to 100 ml). At the end of the addition, the pH of the reaction mixture is 6.8. This mixture is allowed to warm to room temperature and stirring is continued for 3.25 hours. 300 ml of ice-cold water are then added added, the mixture is brought to pH 3 with ice-cold 1N hydrochloric acid and extracted with five 100 ml portions of methylene chloride; the combined extracts are washed twice with water (100 ml portions) and dried over anhydrous sodium sulfate. * Evaporation under reduced pressure at 40 ^° C. gives a mobile liquid which contains a considerable amount of DMSO. It is redissolved in 60 ml of chloroform and the solution is washed 10 times with water (10 ml portions). After drying and evaporation, 0 is obtained. 59 g of a crystalline, white solid from the chloroform phase. It is recrystallized from a mixture of chloroform and petroleum ether, a total of 350 mg (73 %) of short white needles with a melting point of 168 to 174 ° C. being obtained with decomposition.

Analysis C

55 C. : .3 Ή 8th N Cl 02 ber.:.-. 54 , 05 V 3 , 76 8th , 02 10, 53 found: , 65 .79: , 46 10,

The infrared spectrum shows peaks in the carbonyl region at 5.52, 5, 60, 5.80 and 5.85 / U. _: - \

The NMR spectrum shows peaks at 2.13 (4H), 3.78 (1H, d, ■ J = 4.1 Hz), 4.29 (1H, d, J = 4.1 Hz), _f 7.67 (3H), 7.69 (3H) The UV spectrum shows \ fJ2 ^H of 236 (7400), 296 (1800).

- 44a -

5 0 9 8 2 0/1161

The connection has the structure

H H Cl

■ SS Ώ "/

COOH

Conversion of the acid into the methyl ester

200 mg (0.57 mmol) of the acid is dissolved in 30 ml of a 1: 1 mixture of methylene chloride and ether. The solution is cooled and treated with excess ethereal diazomethane. The resulting solution is stirred for 0.5 hour, allowed to warm to room temperature for a further hour and evaporated at 40 ° C., a white crystalline residue being formed. Uinkristallisation from a mixture of chloroform and petroleum ether gives 0.20 g (97%) of colorless needles melting at -feiner 178 to 18O ⁰ C with decomposition.

The connection owns the. structure

COgMe

- 45 -

509820/1161

Conversion of the acid chloride into the methyl ester

500 mg (1.43 mmol) of the crystalline acid chloride is dissolved in 10 ml of methylene chloride and 10 ml of absolute methanol are added. The mixture is stirred at room temperature for 30 minutes, then with 50 ml of methylene chloride. diluted and one after the other with 10 ml of ice-cold 5% bicarbonate solution and washed water. After drying over anhydrous magnesium sulfate and removing the solvent a crystalline residue is obtained. It is recrystallized from methylene chloride / petroleum ether, being 463 mg (93%) of the methyl ester which is identical to the is the methyl ester described above, can be obtained.

analysis

C. 3 4, H 7th N 9 Cl % ber .: 56, 58 4, 16 7th , 71 9 , 78 % found: 56, 37: , 55 , 82

The NMR spectrum shows peaks at 2.10 (m, 4H), 3.77 (1H, d, J = 4.1 Hz), 4.23 (1H, d, J = 4.1 Hz), 6.18 (3H), 7.57 (3H), 7.66 (3H). ■

- 46 -

509820/1161

Direct conversion of anhydro-6-pftthalimidop.enicillin into a mixture of methyl ^ - ^ 'R-cmor ^' S-phthalimido ^ '- oxo) -azetidinyl-3-methyl-2-butenoate and its 2 * S-epimers

Chlorine gas is passed into a solution of 1.0 g (3.04 mmol) of anhydro-6-phthalimidopenicillin in 30 ml of methylene chloride at room temperature. The introduction of gas is ended after 3 minutes and after a further 5 minutes the solvent is removed at 30 ° C. under reduced pressure. The solid residue is redissolved in 20 ml of methylene chlorid and 20 ml of absolute methanol is added. This solution is stirred at room temperature for 30 minutes and evaporated to dryness ^{at 40 ° C. under reduced pressure.} The residue is crystallized from chloroform / petroleum ether, 600 mg (55 %) of the ester having the 2 »R-ChI or configuration being obtained. The mother liquor is chromatographed over a 1.5 × 14 cm aluminum oxide column (Woelm, activity level II). Eluting with 1: -1 methylene chloride / benzene provides 202 mg (18 %) of a new compound in the first 25 ml, followed by 93 mg of a mixture of the 2'R-ester and the new compound in the next 25 ml. Eluting with 40 ml of methylene chloride then provides an additional 68 mg of the 2'R ester.

The new compound is crystallized from methylene chloride / petroleum ether to give the ester with .the 2 ¹ S-chloro configuration is obtained with melting point 192-194 ° C.

,Analysis

C. 3 4, H 7, N 9, Cl ber. ι 56, 42 4, 16 7, 71 9, 78 found: 56, 43 68 60

The NMR spectrum shows peaks at 2.17 (4H, m), 3.77 (1H, d, J = 2.0 Hz), 4.4 (1H, d, J = 2.0 Hz), 6.17 (3H), 7.7 (3H), 7.92 (3H).

- 47 -

509820/1161

m / 12860 _VS 2356852

The structure of this compound is

Stereo selectivity of the chlorination of anhydro-6- phthalimidopenicillin

As already described, 0.5 g (1.52 mmol) of anhydro-6-phthalimidopenicillin are used chlorinated in methylene chloride. That NMR spectrum of the total product shows peaks at 2.12 (4H, m), 3.78 (0.6H, d, J = 4.2 Hz), 3.77 (0.4H, d, J = 1.8 Hz),. 4.25 (0.6H, · d, J = '4.2 Hz), 4.33 (0.4H, d, J = 1.8 Hz), 7.57 (1.8H), 7.65 '(1.2H), 7.68 (1.2H), 7.77 (1

It follows from this that the chlorination reaction proceeds stereoselectively ^{under these conditions, a 60:40 mixture of the 2 f} R-chlorine: 2'S-chlorine epimers being obtained under these conditions. . ·

Crystallization of the product. Methyl en chloride / petroleum ether provides 300 mg (38 %) of the pure dichloride with the 2'R configuration. The mother liquor is then dissolved in 10 ml of absolute methanol and the solution is left to stand for 30 minutes at room temperature with occasional shaking. It is then diluted with methylene chloride, and successively with ice-cold sodium bicarbonate solution and

509 820/1161

Washed with water and dried over anhydrous magnesium sulfate. Evaporation of the solvent gives a residue whose NMR spectrum shows that it consists of a 1: 2 mixture of the 2 ¹ R and 2'S methyl esters. It is chromatographed on aluminum oxide (oil, activity level II). Eluting with 1: 1 benzene / methylene chloride provides 120 mg of the 2'S methyl ester in the first 10 ml of the eluate, followed by a mixture of the 2 ¹ R and 2'S esters. The first eluate is crystallized from methylene chloride / petroleum ether, 55 mg (7 %) of pure 2'S-ester being obtained.

In a second attempt, the. Chlorination carried out as already described for 3 minutes and the solution is left to stand at room temperature for 3 hours. Evaporation yields a mixture containing 68 % 2- (2 ^l S-chloro-3 '| -phthalimido-4 ¹ -oxo) · azetidinyl-3-methyl-2-butenoyl chloride and 32 % of the 2'R-epimer , ie under these conditions the cis-trans ratio is improved in favor of the trans isomer. The same 68:32 ratio is obtained if the chlorination is carried out for 30 seconds and the solution is immediately evaporated to dryness.

If the chlorination is carried out for 10 seconds with a slow stream of chlorine, and the mixture is immediately evaporated to dryness, a single new compound is obtained. The NMR spectrum of this new compound shows peaks at (CDCl ₃ ) 2.1 (4H, d), 401 (2H, s), 7.60 (3H), 7.64 (3H), In CD ₃ COCD ₃ shows the * two protons' singlet at 4.01 a slight splitting to a doublet. The connection becomes the structure

- 49 -. 509820/1161

M / 12860

COSOl

assigned.

Their conversion into the 2 ¹ R- and 2 ¹ S-DiChIOrIdB on further chlorination shows that the compound is an intermediate product in the chlorination of anhydro-penicillin.

- 50

509820/1161

M / 12860

Equilibration of methyl 2- (2 ¹ R-ChIOr-J ^f S-phthalimido-4'-oxo) -acetidinyl-3-methyl-2-butenoate and its 2'S-epimer

A. Starting from the 2 ¹ R isomers

100 mg (0.28 mmol) of the pure 2'R-ester becomes one ■ warm solution of 95.2 mg of tetraethylammonium chloride (0.57 mmol; two molar equivalents) in 15 ml of acetone ^ give. The solution is refluxed for 2 hours, cooled to room temperature and poured onto a mixture of water and ice. Extraction with methylene chloride, followed by washing the extract with water, drying over anhydrous magnesium sulfate and evaporation to provide a crystalline residue. The NMR spectrum of this residue is identical to that of the starting material with the exception that a faint doublet X4.4 (J = 2 Hz) occurs.

The substance is therefore redissolved in 15 ml of acetone containing 95.2 mg of tetraethylammonium chloride and refluxing is continued for a further 14 hours. Isolation in the manner described above gives a residue whose NMR spectrum shows that it consists of a 1: 1 mixture of the 2 ¹ R and 2 ^f S esters.

The experiment is repeated with 100 mg (0.28 mmol) of pure 2'R ester and 238 mg (5 molar equivalents) of tetraethylammonium chloride in 10 ml of methyl ethyl ketone. After refluxing for 4 hours, the product is isolated and determined to be a 3: 2 mixture of the 2 ¹ R and 2'S esters. This ratio does not change if the material is refluxed for a further 8.5 hours in methyl ethyl ketone, which contains 5 molar equivalents of tetraethylammonium chloride.

• - 51 509820/1161

M / 12860

The experiment is carried out with 100 mg of pure 2'R-ester in 15 ml Acetone, the 5 molar equivalents of tetraethylammonium chloride contains, repeated. After 12 hours and after 24 hours of refluxing, a 3: 2 2 · S: 2 · R ratio becomes the ester observed. The product of a 12 hour Reaction to aluminum oxide (Woelm, activity

l -j

stage II) chromatographed. Elute with Benzene / Methylene chloride (1: 1) - as already described - delivers 40 mg of the 2'S compound in the first 25 ml, followed of 3.4 mg of the 2 · S compound in the next 45 ml Eluate.

After refluxing for 6 hours in 30 ml of chloroform containing 5 molar equivalents of tetraethylammonium chloride, 430 mg of the 2'R ester are converted into a 2: 1 2 ^f R: 2 ^f S mixture. , '

B) Starting from 2 ^{t of} S isomers . ·

40 mg of the 2'S-ester for 12 hours in a mixture of 10 ml of acetone and 5 ml of methylene chloride, the 2 molar equivalents Contains tetraethylammonium chloride, kept under reflux. The product is a 1: 2 ^ 'R ^' S mixture.

- 52 -

509820/1161

M / 12860

Chlorination of anhydro-6-phthalimidopenicillin in the presence of tetraethylanunonium chloride '

A solution of 100 mg (0.30 mmol) of anhydro-6-phthalimidopenicillin in 5 ml of methylene chloride containing 252 mg (5 molar equivalents) of tetraethylammonium chloride is chlorinated in the usual way. Evaporation of the solvent gives a residue which consists of a 3: 2 mixture of the 2'S: 2 ^f R-dichlorides.

The experiment is carried out using 10 ml of methylene chloride "is repeated for the reaction. After removing the solvent, 5 ml of absolute methanoi and 5 ml Methylene chloride added and the mixture is 30 min. touched. It is then washed with ice-cold sodium bicarbonate solution and water and dried over magnesium sulfate and evaporated. The residue (132 mg) is a 1: 1 mixture of the methyl esters.

»
The chlorination of the anhydropenicillin in the presence of added chloride ions thus causes a slight increase in the 2'S: 2 ^J R ratioJ

Attempted reaction of 2- (2 ¹ R-ChIOr ^ 'S-phthalimido-4'- oxo) -azetidinyl-3-methyl-2-butenoic acid with sodium azide

50 mg (0.14 mmol) of the 2'R acid is added to a solution of 10 mg of sodium azide (0.15 mmol) in 10 ml of water. After adding a few drops of 5% bicarbonate solution, complete dissolution is achieved. The solution is heated on the steam bath for 4 hours, during which time it turns a dark orange color. After cooling, the solution is brought to a pH of 4 with 1N hydrochloric acid.

- 53 509820/1161

M / 12860 -

and extracted with methylene chloride. The extract is washed with water, dried over anhydrous magnesium sulfate and evaporated to give 24 mg of material. The IR spectrum of this material shows no β-lactam absorption at 5.6 / U. . ; ^; . ; "■ '.-. ■:, ....

In a second experiment, a solution of 20.5 mg (0.33 mmol) sodium azide prepared in 30 ml dimethylformamide, indem.man briefly warmed up on the steam bath and 100 mg "Her 2'R acid (0.29 mmol) is added." The received The solution is heated for 19 hours on the steam bath, cooled, with Water, diluted with 1'ri hydrochloric acid to pH brought from 4 and extracted with methylene chloride. After this Washing with water and drying over anhydrous magnesium sulfate, the methylene chloride is evaporated, whereby 47 mg residue accumulates «A small amount of crystalline Material is used in treating this residue with Obtained methylene chloride / petroleum ether. The IR spectrum of this material shows no azide peak at 4.7 / U and only a phthalimido absorption at 5.62 / U. Evaporation of the Mother liquor gives a residue, its infrared spectrum a weak azide peak at 4.72 / U and something shows β-lactam absorption at 5.60 / U.

Conversion of methyl 2- (2'R-Chror-3'S-phthalimido-4 ¹ -oxo -) - azetidinyl-3-methyl-2-butenoate into methyl 2- (2 * S-azido-3'S-phthalimido-4 '-oxo) -azetidinyl-3-methyl-2-butenoate.

By briefly heating on the steam bath, 10.1 g (0.16 mmol) of sodium azide are dissolved in 15 ml of dimethylformamide and 50 mg of the methyl ester (0.14 'mmol) with the 2 ^l R-chlorine configuration are added to this solution. The resulting solution is stirred at room temperature for 24 hours and then

- 54 -

5098 20/1161

M / 12860

diluted with water and extracted with methylene chloride. The methylene chloride extract is washed thoroughly with water, dried over anhydrous magnesium sulfate and evaporated to dryness. The white crystalline pesticide obtained is recrystallized from chloroform / petroleum ether, 42 mg of unreacted 2'R-ester being obtained.

In a second experiment, the reaction mixture is heated on the steam bath for 5 hours, during which time the 5 color becomes deep yellowish orange. Isolation of the product - as described above - provides 38 mg of a solid residue, the IR spectrum of which is azide absorption at 4.7 / U and Shows carbonyl peaks at 5.58, 5.62, 5.79 and 5.82 / U. That NMR spectrum shows 1-proton absorptions in the β-lactam region at 4.22 and 4.71 (d, J = 2.0).

In a third experiment, 20.2 mg (0.31 mmol) of sodium azide are used in 10 ml of freshly distilled dimethylformamide dissolved, 100 mg (0.28 mmol) of the 2'R-ester are added and the solution is heated on the steam bath for 5 hours. That Product is isolated in the usual manner and is crystallized from 2-butanone / ethanol, with 25 mg of 2'S-azidoester (first crystal fraction) from melting point 142 to t44. C. «,

Analysis C ₁₇ H ₁₅ N ₅ O ₅ :

C H N

calc .: 55.29! 4.09 18.97% found: 55.45 4.02 18.68 %

The NMR spectrum shows peaks at 2.22 (4H, d), 4.22 (1H, d, J = 2.0 Hz), 4.71 (IH, d, J = 2.0 Hz), 6, 17 (3H), 7.70 (3H), 7.91 (3H).

- 55 -

509820/1161

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The structure of the connection is

• O O

CO ₂ Me

In a further experiment, 20.2 mg (0.28 mmol) of sodium azide are dissolved in 10 ml of freshly distilled dimethylformamide by heating on the steam bath for 0.5 hours. 100 mg (0.28 mmol) of 2'R-chloroester are then added, followed by an additional 20.2 mg of sodium azide. The resulting mixture is heated under stirring for 3 hrs. At 9O ⁰ C. Isolation - as described above - yields a semi-solid residue which weighs 68.2 mg (6%) and whose MR spectrum is identical to that described above. Chromatography over a 1x2 column with aluminum oxide (Woelm, activity level II) and elution with methylene chloride leads to 62 mg of crystalline material. Recrystallization from Methylenchiοrid / petroleum ether yields in the first fraction 38 mg of colorless prisms of melting point 144-145 ⁰ C.

- 56 -

509 & 20/1161

M / 12860

Conversion of methyl 2- (2 ^I S-chloro-3 ^l S-phthalimido-4 ^I -oxo) -azetidinyl-3-methyl-2-butenoate into methyl 2- (2'R-azido-3 ¹ S- phthalimido-4 ^l -oxo) -azetidinyl-5-methyl-2-butenoate

100 mg (0.28 mmol) of the pure 2'S chloroester v / earth with 2 molar equivalents of sodium azide in 10 ml of dimethylformamide, as implemented in the experiment described above. insulation in the same way gives 52.2 mg of material. This material is on a 4.5 x 1 cm column with aluminum oxide (Woelm, activity level II) chromatographed. Elute with methylene chloride provides no material in the first 10 ml, followed by 13.0 and 11.1 mg of crystalline material in the next 10 milliliters. Both residues have the same NMR spectrum. You will therefore combined and recirculated from chloroform / petroleum ether, being 19 mg of colorless needles from melting point. 183 to 187 ° C (Decomposition) can be obtained.

The NMR spectrum shows peaks at 2.10 (4H, d), 4.18 (1H, d, 4.0 Hz), 4.33 (1H, d, 4.0 Hz), 6.10 "(3H), 7.47 (6H).

analysis

55 C. 4th H 18th N ber .: .. 55 , 29 4th , 09 19th , 97 ge f .: , 06 , 25 , 10

The structure of this connection is:

- 57 -

509820/1161

M / 12860

Chlorination of the p-toluenesulfonic acid salt of anhydro-6-aminopenicillin . ""'' . '

328 mg (0.89 mmol) of the salt are suspended in 30 ml of methylene chloride, the mixture is ^{cooled to 0} ° C. and gaseous chlorine is passed in for 15 seconds. Subsequently, the resulting yellow solution is 15 min. dry nitrogen is passed in to remove excess chlorine and the solution is evaporated to dryness. Trituration of the residue with dry ether gives a quantitative yield of the mixture of the two compounds shown below:. '

Ts OH

H H

TsOH-

COCl

CQCl

2 ¹ R isomer

2'S isomer

The mixture is a stable (below 20 ^° C.) white powder with a melting point of 148 to 149 ^° C. with decomposition. The IR spectrum shows peaks at 5.55 and [5.62 / U. The 2 ¹ R isomer (formed in 80% yield) shows NMR absorptions (CDCl ₃ ) at ^ 2.20 (2H, d), 2.80 (2H, d), 4.02 (1H, d, 4.2 Hz), 4.82 (1H, d, 4.2 Hz), 7.63 (3H), 7.80 (3H), 8.00 (3H). (The NH ₂ protons are not observed). The 2 ^! S-isomers (formed in 20% yield) shows NMR absorptions at 2.20 (2H, d), 2.80 (2H, d), 4.82 (2H, m), 7.63 (3H), 7.83 (3H), 8.00 (3H).

'- 58 -

509820/1161

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The chlorination of anhydro-6-tritylamino-penicillin, the precursor of the p-toluenesulfonic acid _{salt, does not succeed using Cl 2} , SO ₂ Cl ₂ , pyrrolidone hydrotrichloride, pyridinium trichloride, PCl ₅ and (CgH ₅ O) ₅ PCl. In each case a reaction of the desired type occurs, but the trityl group is lost and the product decomposes.

The chlorination of the p-toluenesulfonic acid salt in the presence of excess tetraethylammonium chloride does not change the ratio of the 2 ¹ R and 2 ¹ S isomers.

Reaction of the chlorination product of the p-toluenesulfonic acid salt of anhydro-6-aminopenicillin with methanol

3.80 g (10.3 mmol) of Anhydropenicillinsalzes are suspended in 150 ml of methylene chloride, the mixture is cooled to -5 ⁰ C and a slow stream of dry nitrogen is passed. Gaseous chlorine is passed into the mixture for 30 seconds and the nitrogen flow is continued for 15 minutes. 75 ml of methanol are added and the yellow solution obtained is at O ⁰ C for 20 minutes with stick

fabric rinsed. Now, the cooling bath is removed and after 15 min. The solvent under reduced pressure at 25 ⁰ C is removed. The residue is treated with 5 ml of methylene chloride and the resulting solution is evaporated again; This procedure is repeated three times, at which point the pale yellow foam no longer smells of chlorine or hydrogen chloride. The foam is dissolved in dry acetone, the orange-colored solution is filtered to remove a small amount of insoluble material to be removed and petroleum ether (30 to 6O ⁰ C) is added to the cloud point. You then add a few drops of acetone to obtain a clear solution and leaves at 10 ⁰ C critical

- 59 -

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crystallize * The white crystals obtained in this way are washed with cold 1: 1 acetone / petroleum ether and dried in vacuo. The yield is 2.7 g (6 %) ■ · Melting point = 125 to 130 ⁰ C (decomposition) «The compound has the following structure:

HH

TsOH.

CO ₂ CH ₅

analysis

47 C- 5, H- ber .: 47 , 48 ■■ 5, 23 ge f .: , 62 54

The NMR spectrum shows peaks at 2.22 (2H, d), 2.90 (2H, d), 4.02 (1H, d, 4.0 Hz), 4.87 (1H, d, 4.0 Hz), 6.48 7.65 DH), 7.78 (3H), 8.15 (3H).

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Conversion of the methyl esters of the 2'R-chloro-3 ¹ S-aminotοsylats into the 2'R-chloro-3'S-phthalimido series

The crude methyl ester obtained by chlorination of 193 mg (0.48 mmol) of p-toluenesulfonic acid salt of anhydro-6-aminopenicillin and subsequent treatment with methanol is dissolved in methylene chloride. The solution is ^{cooled to 0} ° C. and shaken with an ice-cold 5% sodium bicarbonate solution. The colorless organic layer is dried over anhydrous magnesium sulfate, the volume is reduced to 5 ml and 100 mg of N-carbethoxyphthalimide (0.46 mmol) is added, the reaction mixture is allowed to stand at room temperature overnight and the solvent is then removed. The residue is chromatographed on a column with aluminum oxide (Woelm, neutral, activity level II). Elution with 1: 1 benzene / methylene chloride gives 37 mg of undefined material, followed by 68 mg (after recrystallization from chloroform / petroleum ether) methyl ester of 2- (2 · R-chloro-3 ^f S-phthalimido-4 ¹ -oxo) -azetidinyl -3-methyl-2-butenoic acid _f which is identical in every way to the compound obtained from anhydro-6-phthalimido-penicillin.

The sequence of reactions just described is as follows:

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TsOH-H ₂ N-

- J * 5 5

NaHCO-

H- N "" "i

-N

GO ₂ CH ₃ ■

• N.

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Hydrolysis of the chlorination product of the p-toluenesulfonic acid salt of anhydro-a-aminopenicillin -

200 mg of the salt are chlorinated in the usual way and, after the solvent has been removed, the residue is kept under high vacuum for 30 minutes. It is then dissolved in a mixture of 2 ml of D ₂ O and 2 ml of CD ₃ COCD ₃ , cooled to ⁰ ° C. and the NMR spectrum is recorded at intervals. The hydrolysis is over after 3 hours and the solvent is therefore removed by lyophilization. The foam obtained is redissolved in DpO. The NMR spectrum of this solution shows that quantitative conversion into a 4: 1 mixture of 2'R-: 2 ^I S-chloric acids has been achieved.

HH TsOH-H ₂ NJ- ~

1 cl

D ₂ O-CD ₃ COCD ₃ TsOD-

COCl

2'R-chloric acid

TsOH-H ₂ N

COCl

. 5
D O-CD COCD TsOD> D ₂ N-

COOD

■ - 63 -

2'S-hydrochloric acid

509820/1161

Μ / 12,860 ξ $

The 2'R-chloric acid shows NMR absorptions at 2 ^ 37 (2H, d), 2.78 (2H, d), 3.78 (1H, d, 4.0 Hz), 4.93 (1H ,. d, 4.0 Hz), 7.77 (3H), 7.88 (3H), 8.09 (3H). The epimeric 2VS chloric acid shows NMR absorptions at 2.37 (2H, d), 2.78 (2H, d), 4.53 (IH, d, 2.0 Hz), 4.57 (in, d, 2.0 Hz), 7.77 (3H), 7.98 (3H), 8.24 (3H). ' \\: ■ - ■ - ■ -., -

Methyl ester of 2- (2 ^l R-chloro-3'S-amino-4 ^f -oxo) -azetidinyl-3-methyl-2-butenoic acid

The methyl ester of the 2 ^l R-chloro-3 ^l S-aminotosylate is, as already described, neutralized with bicarbonate. Evaporation of the methylene chloride solution yields the free base as a crystalline compound with a melting point of 67 to 70 ° C. with decomposition. The compound is not stable in the solid state and becomes noticeably dark after 10 "minutes at room temperature.

The NMR spectrum of the compound shows peaks at 3.97 (1H, d, 4.1 Hz), 5.38 (1H, d, 4.1 Hz), 6 ^ 22 (3H), 7.70 (3H), 8.00 (3H). The position of the NHp protons varies over time; although these peaks shift, there are no further changes in the spectrum. The IR spectrum of the compound shows peaks at 2.94, 5.60, 5.80, 6.12 / U which (in ) do not change over time.

The structure of this compound is

'- I S

COOCB

-64 -

50982Ό / 116Τ

M / 12860

Methyl ester of 2- (2'S-ChIOr ^ 'S-amino-4'-oxo) -azetidinyl-3-methyl-2-butenoic acid by epimerization of the 2 ¹ R compound

440 mg (1.09 mmol) of 2 ^f R-chloro-3'S-aminotosylate and 1.387 g (9.1 mmol) of tetramethylguanidinium chloride (9 molar equivalents) are refluxed in 9 ml of chloroform (spectroscopic quality) for 4.5 hours . The solution is then ^{cooled to 0 °} C., washed with ice-cold 5% strength bicarbonate solution, dried over anhydrous magnesium sulfate and evaporated to dryness ^{at 25 ° C. under reduced pressure.} The NMR spectrum of the residue indicates that there is a 5: 1 mixture of the 2 ¹ S and 2'H epimers, the predominant isomer being the 2 ¹ S compound, the structure of which is as follows

• H H

is present.

The NMR spectrum of this compound shows peaks at 4.45 (1H, d, 1.8 Hz), 5.63 (1H, d, 1.8 Hz), 6.18 (3H), 7.70 (3H) , 8.00 (3H). The position of the NH ₂ protons depends on the age of the solution.

Complete separation of the two epimers can be achieved by chromatography with aluminum oxide and eluting with 1: 1 benzene / ethyl acetate. However, it is generally because of the instability of these compounds more convenient to separate at a later stage.

- 65 5 0 9 8 2 0/1161

2356062.

M / 12860

Conversion of the p-toluenesulfonic acid salt of methyl-2-r- (2'R-chloro-3'S-amino-4'-oxo) -azetidinyl-3-diethyl-2-butenpat into the 2 ^J S-azido-3 ⁱ S- aiainoester '- .; _: .-

448 mg (1.1 mmol) of the crystalline methyl ester ¹ with the '2'R-ChIOr-J ¹ S-TsOH-H ₂ N configuration and 347 mg of' tetramethylguanidinium azide (2.2 mmol, 2 molar equivalents) - .. are refluxed for 75 minutes in 20 ml of anhydrous chloroform. The solution is allowed to cool and an excess of anhydrous ether is added to precipitate tetramethylguanidinium chloride, which is filtered off. After removing the solvent, the material obtained is the desired 2 ^l S-azido ^ 3 ^l S-amino ester (237 mg, - 90 %) , which is pure according to thin-layer chromatography and NtIR spectrum. - 'The NMR spectrum shows peaks at 4.97 (IH, d, 1.8 Hz),' 5.87 (1H, d, 1.8 Hz), 6.20 (3H), 7.00 (2H >., '7.72 (3H), 8.02 (3H). ·' _ ■ - ^: ·

The connection has the structure ..

JT.

COOCH ₃

The compound can also be removed by washing the chloroform reaction mixture isolated with cold water, drying and evaporation. Tetraethylammonium azide (in various Solvents) and sodium azide (in dimethylformamide) less effective in this reaction * ..

-66 -

509820 / T161

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Conversion of the p-toluenesulfonic acid salt of methyl 2- (2'S-chloro-3 'S-amino-4 ^l -oxo) -acetidinyl-3-methyl-2-butenoate into the 2 ^f R-azido-3'S-aminomethyl ester

532 mg (2.29 mmol) of a 4: 1 mixture of the 2 ¹ S- and 2 ¹ R-ChIOr-3'S-amino esters from an epimerization of the 2'R-chlorine compound and 500 mg (3.2 mmol) of tetramethylguanidinium azide are in 10 ml of pure chloroform kept under reflux for 4.5 hours. The mixture is then washed with ice-cold 5% strength bicarbonate solution, dried over anhydrous magnesium sulfate and evaporated, with 465 mg (85 %) of a 3.3: 1 mixture of 2 ¹ R and 2 ^t S-azido-3 ^l S -aminomethyl ester can be obtained. This mixture is separated by chromatography on neutral aluminum oxide (Woelm, activity level II). Eluting with benzene: ethyl acetate (2: 3) gives the pure 2 ¹ R-azido-3'S-amino ester as a crystalline compound. Recrystallization from chloroform / petroleum ether gave long needles of melting point 116-117 ⁰ C.

analysis

C. 5 H N % ber .: 45, ^ 9 5 , 48 29.28 % found: 45.42 , 24 28.81

The IR spectrum shows peaks at 4.72, 5.62 and 5.80, and the like. That NMR spectrum shows peaks at 3.97 OH, d, 4.1 Hz), 4.38 (d, 4.1 Hz), 6.22 (3, H), 7.60 (2H), 7.70 (3H), 8.00 (3H).

The connection has the structure

• '■ HH

COOC

«3

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is

Separation of the 2 ¹ S and 2 ^l R-azido-3'S aminomethylester on the p-toluenesulfonic

239 mg of a 1: 1 mixture of the 2 ¹ S- and 2'R-azido compounds are dissolved in 3 ml of dry acetone and the solution is treated with an equivalent amount of p-toluenesulfonic acid hydrate. Crystallization begins after 5 minutes. After 15 hours 1 ml of acetone are added and after a further 20 minutes 5 ml of ether are added * Subsequent filtration yields 150 mg. "(72 % recovery, based on an isomer) of a crystalline p-toluenesulfonic acid salt. The regeneration of the free base (NaHCO ^ - ^ O-CHgGl ^) shows that it is the salt of the 2'S-azido compound. The separation method is as follows ^:;

H ^! H

HH

OOCB,

TsOH-H ₂ O

TS OH

COOCH

COOCH

COOCH

less soluble

more soluble

- 68 -

SO 9820/1 161

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JO

Phenylacetylation of methyl 2- (2 ^t R-chloro-3 ^t S-amino-4 ^l- oxo) · azetidinyl-3-methyl-2-butenoate

The obtained from 722 mg (1.78 mmol) of p-toluenesulfonic acid-free 2-chloro-3 ^I R ^l S-amino ester is dissolved in 15 ml of methylene chloride and 242 mg (1.78 mmol) of phenylacetic acid are added. This solution is treated dropwise with stirring with a solution of 405 mg (1.96 mmol) of dicyclohexylcarbodiimide. Dicyclohexylurea precipitates out before the addition is complete. The reaction mixture is left to stand for 3 hours at room temperature; after this time there is no longer any free amine present (by thin-layer chromatography). Most of the urea is removed by filtration and the filtrate is washed with ice-cold 5% bicarbonate solution. After drying over anhydrous magnesium sulfate, the organic phase is evaporated to give a solid residue. It is made in boiling chloroform. dissolved and an equal volume of petroleum ether (30. to 60 ° cJ is added, whereupon crystallization occurs. The crude product weighs 700 mg. A further recrystallization from the same solvent mixture gives 500 mg (80 %) of methyl-2- (2 · R-chloro- 3'S-phenylacetamido-4'-oxo) -azetidinyl-3-methyl-2-butenoate from melting point 111 to 15 ° C. with decomposition. The NMR spectrum of this compound shows peaks at 2.62. (5H), 3.10 ( 1H, d, 10 Hz), 3.94. (1H, d, 4.2 Hz), 4.34 (1H, q, 4.2, 10 Hz), 6.22 (3H), 6.31 ( 2H), 7.70 (3H), 8.02 (3H) The same compound can be obtained, but in somewhat lower yield, using diisopropylcarbodiimide or phenylacetyl chloride and triethylamine The compound has the structure

I §

PhCH ₂ CONH

• N.

'CO ₂ CH,

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50 9 8 2 0/1161

Μ / 12860 η _Α

Phenylacetylation of methyl 2- (2 ¹ S-chloro-3 'S-amino-4' -oxo) -azetid.inyl-3-methyl-2-butenoate. · ■ · · ·.

Cyclization to the oxazoline. .-

651 mg (1.6 mmol) of the p-toluenesulfonic acid salt of methyl 2- (2 'R-.chlor-S' S-aemi "no-4 '-oxo) -azetidinyl-3-methyl-2-butenoate and. 1.210 g of tetramethylguanidinium chloride (5 molar equivalents) are refluxed in 9 ml of chloroform (spectroscopic quality) for 4.5 hours. The solution obtained is cooled one after the other with ice-cold 5% strength sicarbonate and ice-cold, saturated sodium chloride solution washed, dried over anhydrous magnesium sulfate, and used immediately for the next step (earlier experiments have shown that the above procedure results in a 5 si or 6: 1 excess of the -2-S-chloro-3'S-amino compound over the 2 'R 3 ^1-chloro-S ^ -amino leads). Lqsung to 228 mg (1.7 mmol) of phenylacetic acid, .222 mg (1.8 mmol) and 20 ml Diisopropylcarbodiiffiid Methylenchlörid are added and this is -Losung 1 hr. The removal of the solvent gives a residue whose NMR spectrum peaks at 2.73 (phenyl) _r 4.72 (d, -1 , 9 Hz), 5.33 (d, 1.9 Hz), "6.30 (OCH ₃ ), 6.47 (CH ₂ ), 7.80 - (CH ₃ I, 8.05 (CH ₃ ) -shows. The structure of this compound is

CO ₂ CH ₃

The spectrum also shows peaks at 4.02 (d, 3.8 Hz), 4.80 (d, 3.8 Hz), 6.33 ,. 7.83, 8.43. This is the spectrum of connection with the structure.,

50 9820/1161

M / 12860

CH ₂ Ph

N ₃ 2 _n q

HHM ¹ '

ie 2-Benzyl-6- (1'-methoxycarbonyl-2'-methylprop-i * ~ enyl) -1-oxa-3,6-diaza-4S, 5R-MCyCIo [3.2.0] hept-2-en- 7-on, formed by the intramolecular cyclization of the preceding compound. The cyclization occurs when standing, however, it is obtained by heating a solution of the chlorine compound, by shaking with bicarbonate, or by chromatography on alumina or silica gel one, each of the epimers 2'-chloro'-acylaminoesters, as described below will. .,

Chromatography of the above mixture on silica gel and eluting with petroleum ether / ethyl acetate (1: 1) yields the oxazoline. Recrystallization from ethyl acetate gives 240 mg (49 %, starting from the pj-toluenesulfonic acid salt) with a melting point of 126.5 to 127.degree

The NMR spectrum of the pure compound shows peaks at 2.73 (5H), 4.02 (1H, d, 3.8 Hz), 4.80 (1H, d, 3.8 Hz), 6.30 (3H), 6.33 (2H), 7.83 (3H), 8.43 (3H).

509820/1161

M / 12860

Formation of the oxazoline when chromatographing the cis-2 ^f R-chlorine-2 ^! S-phenylacetamIdo-methyl ester:; ..-. "

100 mg of the crystalline ice compound are on aluminum oxide (¥ oelm, activity level II) chromatographed. Elute with carbon tetrachloride / benzene (1: 1) gives 38 mg of the oxazoline in the first fractions, followed by the non-cyclized cis compound. ^

In a second experiment, 1.0 g; the ice connection chromatographed on 37 g of silica gel and eluting with 1: 1 petroleum ether (30 to 60 ° C): ethyl acetate carried out. Man receives in successive fractions 450 mg of recovered eis compound, 140 mg of a mixture of the eis compound and of oxazoline and 210 mg of oxazoline. The implementation is as follows:

PhCH ₂ CONHl, H H rf ⁰¹ ' 0 CO ₂ CH ₃

or

SiO,

2H ₂ Ph

CO ₂ CH ₅

In a third experiment, 157 mg (0.68 mmol) of the crystalline 2 »R-chloro-3 ^l S-aminomethyl ester in 8 ml of dry methylene chloride are successively mixed with 92 mg of phenylacetic acid (0.68 mmol) and 93 mg of diisopropylcarbodiimide (0.74 mmol). The solution and / the precipitated diisopropylurea is stirred for 22 hours at room temperature. is filtered off. The addition of ether to the mother liquor completes the precipitation of the urea. Evaporation

- 72 -

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of the filtrate, followed by chromatography on a 1.0 χ 7 cm column with aluminum oxide (activity level II) with 1: 1 benzene / methylene chloride, gives 83 mg of crystalline material. Evidence from NMR and thin layer chromatography that it is a mixture of oxazoline and - ^ 'S-phenylacetamidoester.

Isolation of the methyl 2- (2 ^r R-phenylacetoxy-3'S-phenylacet-_ amido-4'-oxo) -azetidinyl-3-methyl-2-butenoate and the oxazoline following the phenylacetylation of the epimeric methyl-2- ( 2 '-chlor-3 ¹ S-amino-4 ^I -oxo) -azetidinyl-3-Iaeth.yl-2-buΐenoate ~

4.428 g (10.9 mmol) of the p-toluenesulfonic acid salt of methyl 2- (2'R-chloro-3 'S-amino.-4 ^f -oxo) -azetidinyl-3-methyl-2-butenoate and 8.260 g ( 54.5 mmol) of freshly prepared tetramethylguanidinium chloride are kept under reflux in 65 ml of chloroform (spectroscopic quality) for 2 hours. The dip-cooled solution is washed successively with ice-cold 5% bicarbonate solution and ice-cold, saturated sodium chloride solution and then dried over anhydrous magnesium sulfate. A solution of 1.490 g (11 mmol) of phenylacetic acid and 1.500 g (11.9 mmol) of diisopropylcarbodiimide in 25 ml of methylene chloride is then added and the mixture is refluxed for 2 hours. The NMR spectrum of the reaction product shows only a small amount of the oxazoline (based on the peaks at 7.83 and 8.43). The solution is then washed with ice-cold 5% bicarbonate, dried over anhydrous magnesium sulfate and evaporated. The NMR spectrum of this residue shows that the oxazoline is now the main product. So washing with bicarbonate caused the cyclization. The product will stand at 10 ° C overnight.

- 73 509820/1161

■ / - 2 3568.! r

M / 12860 '-.,; ■

left, then dissolved in methylene GM © ri4 \ again _j - and again - shaken with bicarbonate solution. After the -.- drying ^ and the; Evaporation of the organic layer, a liMR spectrum of the residue is recorded; Ve ^, shows, that the mixture contains 70% oxazolines. The mixture -will in 1: 1 ethyl acetate: IPetrolether by. Filtered fO g of silica gel and carefully chromatographed on 110 g of silica gel. Elution is carried out with graded mixtures of petroleum ether and ethyl acetate (250 ml of 20% ethyl acetate, followed by 200 ml of 30% ethyl acetate, followed by 200 ml of 0% ethyl acetate, followed by 200 ml. 55% of ethyl acetate, followed by 200 ml 45 ml ethyl ^ Äthyläcetat followed _r i vop 50 $ ethyl acetate sodium) durchgeführtj collecting 40 ml fractions, fractions 1 to 3 provide 100 mg of methyl 2- ^(2? R-chloro-3'S phenylacetamido-4-oxo ^r) -azetidinyl-S-inetnyl-e-butenoate. Fractions 5 to 12 provide -1.475 gf of the oxazoline. Fractions 13 to 15 "v / erdon chromatographed again and yield, in addition to an additional 150 mg of oxazoline (a total of 1.625 g _? 48 %), 240 mg of a compound to which the following structure is assigned: ■

; ; PhGH ₂ CONH

CQ ₂ CH ₃

This compound shows IR rAbsorptionen at 2.9, 3.0, 5.60, 5.70, 5.80, 5.98 _S, 6.05; 6 * 65 / U. The NMR spectrum has peaks at 2.65-2.75 (1OH, m), 3.26 (1H _r d, 9.5 Hz), 3.68 (1H, d, 4.1 Hz), 4, 53 (q, 4.1, 9.5 Hz), 6.27 (3H), 6.43 (2H), 6.54 (2H), 7.82 (3H), 8.28 (3H). The mass spectrum shows a molecular ion at m / e 450.

- 74 - ■

Μ / 12860 λ,

Phenylacetylation of "methyl-2- (2- ¹ R-azido-3 ^f S-amino-4 ¹ - ■ · oxo) -azetidiiiyl-3-methyl-2-butenoate

415 mg (1 _r 74 mmoles) of crystalline aminoazide is dissolved in 10 ml of dry methylene chloride and 235 mg of phenylacetic acid (1.73 mmoles) and 220 mg of diisopropylcarbodiimide (1.75 mmoles) are added successively to this solution. The solution is refluxed for 9 hours and the solvent is then removed. The residue is triturated with cold carbon tetrachloride and the material which is soluble in carbon tetrachloride (now freed from the majority of diisopropylurea) is converted to aluminum oxide (Woelm, activity level II ) chromatographed. Eluting with benzene / ethyl acetate yields 641 mg of a pale yellow oil. This is chromatographed again, with 601 mg (97 %) of methyl 2- (2 ¹ R-azido-3 ¹ S-phenylace-tai} iido-4 ^! -Oxo) -azetidinyl-3-methyl-2-butenoate after recrystallization from carbon tetrachloride / petroleum ether in the form of long needles with a melting point of 102 to 103 ° C. The connection has the structure

CO ₂ CH ₃

analysis

57 C. 5, H 19th N about «j 56 , 13 5, 36 19th , 60 found: , 76 35 , 42

- 75 -

509820/1161

H / 12860

The MMR spectrum shows peaks at 2.72 (5H), 3, 38 · (1H, gate), 4.57 (1H, el, 4.2 Hz), 4.72 (1H, d, 4.2 Hz ), 6.27 (3H), 6.38 (2H), 7.77 (3H), 8.05 (3H).

Phenylacetylierung of methyl 2- (2 ^L S-azidö ^3-r S-ainino-4 ¹ - '\ oxo) -azetidinyl-3-methyl ~ 2-butenoate.

323 mg (1.35 mmol) of the amino azide are dissolved in 10 ml of methylene chloride containing 110 mg (1.09 mmol) of triethylamine, and 170 mg (1.1 mmol) of phenyiacetyl chloride in 5 ml of methylene chloride are added dropwise with stirring added * After 30 min. the reaction mixture is washed with five 10 ml portions of water, dried over anhydrous magnesium sulfate and evaporated. Chromatograph on aluminum oxide (Wo.elm,. · Activity II) and elute with. Benzene gives 185 mg of ^ methyl-2- (2 ^I S-azido-3 ^l S-phenylacetamido-4 ^t -oxo) -azetidinyl-3-methyl-2-butenoate in the form of a colorless oil. The connection has the structure :.

HH _w

-N

. CO ₂ CH ₃

The EMR spectrum shows peaks at 2.67 (5H), 3.22 (1-H, d, 8 Hz), 4.73 (1Ή, d, 2.0 Hz), 5.32 (1H, q, 2.0, 8 Hz), 6.25 (3H), 6.38 (2H), 7.73 (3H), 8.00 (3H). .

The same compound is obtained in 83% yield by one uses phenylacetic acid and diisopropylcarbodimide in methylene chloride.

■ - 76 - -

50 98 20/1161

M / 12860 ¹

Conversion of methyl 2- (2 ¹ R-clilor-3'S-phenylacetamido-4'-oxo) -acetidinyl-3-methyl-2-butenoate into methyl 2- (2'S-azido-3'S-phenylacetamido-4'-oxo ) Acetidinyl 3-methyl-2-butenoate

126 mg (0.8 mmol) of tetramethylguanidinium azide in 3 ml of chloroform (spectroscopic quality) is added dropwise with stirring to a solution of 229 mg of the chloride (0.66 " mmol) in 5 ml of chloroform ". After the addition is complete, the reaction mixture is refluxed for 2 hours. At this point there is no longer any unreacted chloride. The reaction solution is washed with water, dried over anhydrous magnesium sulfate and evaporated. The residue is chromatographed on silica gel, 75 mg of the azide identical to that described in the previous experiment are obtained.

The reaction equation is:

H H

PhCH ₂ CONH »

0 =]

NH ₂ N ₃
Me ₂ NC-NMe ₂

H H

PhCH ₂ CONHiH ^

O = J

-N

'CO ₂ CH ₃

CO ₂ CH

- 77 -

509820/1161

M / 12860

Production of te; rt.-M ^ 1 ^ 2- (2'-R- ^ 4 '-oxo) -azetidinyl-3-methyl ^ -butehöät and tert. fButyl-2- (2 · -S-chloro-3 ^r -S-phthalimIdo ~ 4'-oxo) -a2etidinyI - 3-methyl-2-butehoate by reaction of the anhydrö-e-phthalimidopenioillin-chlorination product with tert. ^ Butenol ';'' ■ ''

^; A 2 ^ 1 Mschurig ^; vön-! 2 ^^^

•. ■ azetidinyl-3-nlet] iyi-2-T3ütenoylchlorid and these 2 * -S-epimers

shake overnight at room temperature in methylene chloride ^;

The 5-tertiary Möiäqüiväleht'e Butänoi ehthält.'Eindampfen the '■ solution results thanks to a hybrid, the 15 ^b A tertv butyl ^' ■ ester containing (based on the integration of the ^tert-butoxy: peaks at 8.47 in the NMFt spectrum). The Ümwaiidlung sic'h increased to 40% i vrenh to the mixed acid chloride ⁷ overnight

heated to reflux in methylene chloride / tert-butane oil. Heating in tert-butanol solution for 20 hours ^; medium causes a 75 % conversion into a mixture of tert-butyl ester which ^{: maintains the} same 2: 1 ratio of epimers as the acidic

• chloride precursors. Chromatography on silica gel separates them

_: Connections. The cis isomer is obtained as a colorless oil, its NMR spectrum shows peaks at 2.15 (4H), 3.85 (IH, d, 4 Hz), 4.38 (IH, d, "4IiZ.) , 7.71 (.6H), 8.46

, The compound; has the following formula:

- 78 -

50 9820/1161

so

ι The trans isomer crystallizes on standing. Umkri'stalli-

cation from ethanol gives a material that is rated at 170 to 172

: melts. The NMR spectrum shows peaks at 2.29 (4H, d),

3.85 (IH, d, 2.0 Hz), 4.55 (IH, d, 2.0 Hz), 7.73 (3H),

■ 1.98 (3H), 8.47 (9H).

Analysis C _0n H ₀ -, N _n O ₁ -Cl:

calculated: bound:

This compound has the following formula

C. , 32 H 23 N 91 % 59 , 05 5, 36 6, 75 % 59 5, 6,

COOtBu

Reaction of tert-butyl-2- (2'-R-chloro-3 ^I -S-phthalimido-4 ^I -j oxo) -azetidinyl-3-methyl-2-butenoa ^> t with trifluoroacetic acid

100 mg of the ester are dissolved in 2 ml of trifluoroacetic acid. After two minutes at room temperature, the solvent is removed under reduced pressure. The residue is recrystallized from chloroform / petroleum ether, with 2- (2'-R-chloro-3 ' -S_-phthalimido-4'-oxo) -azetidinyl-S-methyl ^ -butenic acid which is identical in all respects to the compound prepared by direct hydrolysis of the dichloride.

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Allyl bromination of methyl 2- (2'-R-chloro-3'-S ~ phthalimido-4'-oxo) -azetidinyl ^ -methyl - ^ - butenoate

To a suspension of 100 mg (0.276 mmol) of the methyl ester in 7.5 ml of carbon tetrachloride are added 10.8 mg (0.61 mmol) of N-bromo-suecinimide and 2 mg of benzoyl peroxide. When this mixture is heated to reflux, the ester dissolves. »Then the heat source is replaced by a 100 watt PhOt ¹ OfIoOd-LaInPe-No .. with reflector, - which are arranged close to the reaction flask and continue to stir until the reaction is complete ( 10 mins). The resulting suspension is cooled, diluted with a small amount of chloroform, and the precipitated succinimide is removed by filtration. Evaporation of the filtrate gives a white crystalline solid, the NMR spectrum of which shows no absorption in the region of the allylic methyl groups (7.5 to 8.0) but shows succinimide absorption at 7.2. This is partially removed by trituration with carbon tetrachloride and methylene chloride and the entire product is then chromatographed on neutral aluminum oxide (Woelm, activity level II). Elution with methylene chloride results in a crystalline dibromine compound which, after recrystallization from carbon tetrachloride / petroleum ether, melts at 65 ° to 70 ° (Dec.) And weighs 116 mg (80 %).

The compound has the following formula

0 ".

- 80 -

5 0 9 8 2 0/1161

C. 22nd H 52 N 38 39 95 2, 36 5, 40 38, 2, 5,

analysis

calculated:
found: ·

The NMR spectrum shows peaks at 2.13 (4H), 3.80 (IH, d, 4.2 Hz] 4.21 (IH, d, 4.2 Hz), 5.07 (IH, d, 10.2 Hz), 5.23 (IH, d, 10.9 Hz), 5.27 (IH, d, 10.2 Hz), 5-33 (IH, d, 10.9 Hz), 6.11 (3H). The spectrum appears unusually complex because one of the -CHpBr groups is cis to the methoxycarbonyl substituent and the other is in the trans position and the methylene protons in each -CHJ3r group are magnetically non-equivalent. The IR spectrum shows Peaks at 5.55, 5.62 and 5.79 µ.

Allyl bromination of methyl 2- (2'-S-chloro-3'-S-phthalimido- i

4 ^! -oxo) -azetidinyl-3-methyl-2-butenoate. . !

100 mg of the compound are reacted with two molar equivalents of N-bromosuccinimide in exactly the same manner as described above. Chromatography on aluminum oxide (Woelm, activity level II) and eluting with methylene chloride gives 122 mg (84 %) of the desired compound as a clear, viscous oil which is shown to be homogeneous by NMR and thin layer chromatography. The NMR spectrum shows peaks at 2.08 (4H), 3.62 (IH, d, 1.7 Hz), -4j27 (IH, d, 1.7 Hz), x 5.05 (IH, d, 10.0 Hz), 5.36 (IH, d, 10.0 Hz), 5.38 (IH, d, 11.0 Hz), 5.57 (IH, d, 11.0 Hz), 6, 04 (3H). This compound has the following formula

V = H ₂ Br

CO ₂ CH ₃

509820/1161

Allyl bromination of methyl 2- (2'-R-azido-3'-S-phenylacetamido-4'-oxo) -azetidinyl = 3-methyl-2-butenoate

136 mg (0.38 mmol) of the methyl ester, 210 mg (1.18 mmol, 3 molar equivalents) of N-bromosuccinimide and 2 mg of benzoyl peroxide are refluxed in carbon tetrachloride suitable for spectroscopy. The heat source is then removed and replaced by a 100-watt Phötoflood lamp No. 2, which is attached close to the reaction vessel. The lighting and refluxing is stopped after 9 minutes. The resulting suspension is cooled, filtered off and the reddish filtrate is treated with ice-cold 10% strength The resulting yellow solution is dried over anhydrous magnesium sulfate and evaporated to give 143 mg (77 %) of a viscous oil. Chromatography on alumina eluting with benzene and then with chloroform gives 29 mg of material which has the same NMR- Spectrum has like the reaction product, which does not crystallize.This NMR spectrum shows peaks at 2.67 (5H, m), 3.33 (IH, br), 4.32 (IH, d, 4.0 Hz), 4 .50 (IH, d, 4.0H z), 5.38 (2H, sl.br.), 5.63 (2H, sl.br.), 6.17 (3H), 6.18 (2H). The IR spectrum showed peaks at 2.97, 4.71, 5.60, 5775 and 5.95 ^ u. The spectra are consistent with a compound of the following structure;

II H

CO ₂ CH ₃

- 82 - 509820/1161

ti

Reaction of methyl 2- (2 ^! -R-chloro-3'-S-phthalimido-4 ^t -oxo) -azetidinyl-3-methyl-2-butenoate with 1 molar equivalent of N-bromosuccinimide

75 mg (0.207 mmol) of the cis compound, 36.9 mg (0.207 mmol) of N-bromosuccinimide and 1.5 mg of benzoyl peroxide in 5 ml of carbon tetrachloride are heated to reflux until a clear solution is obtained. Then remove the heat source and stir the mixture while using a 100 watt photoflood lamp No. 2 amp. "Illuminated. After 10 minutes the reaction is complete and the mixture is cooled to room temperature and filtered to give 17 mg (85 % of the theoretical amount) of succinimide, which is identified by its melting point and its IR spectrum. The filtrate is evaporated to dryness, to result in a white solid residue, which, as demonstrated by the NMR spectrum a small ^amount. succinimide and exactly one containing 1: 1 mixture of the two possible mono-brominated compounds the solid is chromatographed on alumina, to the.. Succinimide

to remove. Eluting with methylene chloride gives the 1 ..:. 1- Mixture of monobromination compounds as a white foam (87.5 mg, 96 %). The NMR spectrum ΐ of this mixture ’shows peaks at 2.20 (4H), 3.83 (0.5H, d, 4.2 Hz), 3.87 (0.5H, d, 4.2 Hz), 4.28 (0.5H, d, 4.2 Hz), j 4.32 (0.5H, d, 4.2 Hz), 5.10-5.88 (2H, m , overlapping AB quartets), 6> 15 (1.5H), 6.17 (1.5H), 7.55 (3H).

Ά

j These compounds have the following formulas

■ · "■■" HH

- 83 -

509820/1161

^{M / 1286} ° M 235 £ 862

'■ _ Methyl 2- (2' -R-chloro-3V-S ~ phthalimidb-4 '-oxo) -acetidinyl • 3-methyl-4-bromo-trans-2-butenoate and

Methyl 1-2- (2 · -R-chloro-3'-S-phthalimido-4 " ¹ -oxo) -acetidinyl-3-methyl-4-bromo-cis-2-butenoate.

Conversion of the cis and trans methyl 2- (2'-R-chloro-3'-S-phthalimido-4'-oxo) -azetidinyl-3-methyl-4-bromo-2-butenoate into the cis and trans methyl 2- (2'-R-chloro-3'-S'-phthalimido-4'-oxo) -azetidinyl-3-methyl-4-azido-2-butenoates

The 1: 1 mixture of 87.5 mg (0.198 mmol) of monobrominated Compounds treated in 3 ml of chloroform suitable for spectroscopy one with 34.4 mg (0.218 mmol) of tetramethylguanidinium azide. Immediately a pale yellow color results. One stirs the solution for 3 hours at room temperature (under these conditions the 2'-chloro substituent is stable), after which this is pale yellow-brown in color. It is diluted with chloroform, washed with water, decolorized with activated charcoal and dried over anhydrous magnesium sulfate. Evaporation of the solvent! gives 83.9 mg of a white solid residue. investigation this residue by means of thin-layer chromatography does not reveal any unreacted monobrominated compounds.

- 84 -

50 9820/1 161

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i6

The IR spectrum shows peaks at 4.70 (azide), 5.52 (ß-lactam), 5.58 and 5.79 µ (phthalimido), 5.75 (ester) and 6.16 u.

The NMR spectrum shows that the azides have been obtained.

It shows. Peaks at 2.17 (4H), 3.78 (IH, d, 4.0 Hz), 4.22 (IH, d, 4.0 Hz), 5.12 - 5.87 (2H, overlapping AB quartets),

6.12 (3H), 7.58 (3H).

The two isomers obviously have the same thing

NMR spectrum.

The structures of these compounds are.

ι Methyl 2- (2 '-R-chloro-3' -S-phthalimido-4 ^f -oxo) -azetidinyl 3-methyl-4-azido-trans-2-butenoate _>

'■. and

i ?! he

ΛN ... -

• ο .-. Ο ⁴

-N-

• f

"3

CO ₂ CH ₅

j methyl 2- (2 '-R-chloro-3' -S-phthalimido-4 '-oxo) -azetidinyl-3-methyl-4-azido-cis-2-butenoate.

- 85 -

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^{M / 1286} °

235686?

Conversion of the cis and trans methyl 2- (2'-R-chloro-3 ^I -S-phthalimido-4 '-oxo) -azetidinyl-3-methyl-4-azido-2-butenoates into the cis and trans-methyl-2- (2-chloro-3 ^{t -R!} -S-phthalimido-4-oxo ^I) -azetidinyl-3-methyl-4-amino-2-butenoates

The mixture of 83.9 mg of azides in 10 ml of benzene is over

2 ■ 50 mg Adams catalyst at 3.16 kg / cm (45 psi;) 3 hours hydrogenated. The reaction solution is then diluted with chloroform and passed through a pillow of anhydrous magnesium sulfate. "Evaporation at 25 gives a sticky solid residue, the IR spectrum shows a peak at 4.7 μ » about half the intensity of one in the same position as the starting material. The NMR spectrum shows peaks due to " of the azide and also some new peaks. The reduction was therefore incomplete. The hydrogenation will continue for an additional 3 hours

2 "

repeated at 3.16 kg / cm (45 psi). The resulting mixture is diluted with chloroform and passed through Pillows of anhydrous magnesium sulfate. The black filtrate becomes after filtering through "Celite" or after shaking (of an aliquot) with 5% sodium bicarbonate solution does not discolored. Evaporation at room temperature gives a black one solid residue (92 mg), the £ R spectrum of which is a complete Shows disappearance of the azide absorption and its NMR spectrum a disappearance of the multiplets in the range 5 - 6'TT and a Occurrence of new multiplets in the range 7T results. The solid is redissolved in chloroform, treated with activated charcoal and anhydrous magnesium sulfate through a pillow given, resulting in a clear colorless solution. Evaporation gives 87 mg of a solid residue. The NMR spectrum shows peaks at 2.18 (4H), 3.88 (IH, d, 4.1Hz), 4.33 (IH, d, 4.1 Hz), 6.90-7.20 (2H, m), 6.20 (3H), 7.67 (1.5H), 7.70 (1.5H]

j - - 86-

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! M / 12860

The structures of the amino compounds are

'. Methyl 2- (2 '-R-chloro-3' -S-phthalimido-4 '-oxo) -azetidinyl-3-i-methyl-4-amino-trans-2-butenoate

H H. ν

CH

CO ₂ CH ₃

Methyl 2- (2 ^f -R-chloro-3'-S-phthalimido-4'-oxo) -azetidinyl-3-methyl-4-amino-cis-2-butenoate.

- 87 -

509820/1161

ι Cyclization of "methyl-2- (2 ^l -R-chloro-3 ^r -Sp] athalimido - 4 ^r -

! oxo) ~ azetidinyl-3-methyl-4-amino-trans ~ 2-butenoate

; 3-methyl-4-meth.oxycarbonyl-7-S-phthalimido- = lj5-diaza = 6-S-

; bicyclo- [4,2,0] -oct-3-en-8-one '

The above mixture of amines is obtained unchanged after standing at room temperature for 16 hours and Was refluxed for 3 hours in chloroform. ■

The 60.5 mg (0.159 mmol) of mixed amino esters in 10 ml of anhydrous tert-butanol are treated with 17.8 mg (0.159 mmol) of freshly prepared potassium tert-butoxide. The initially pale yellow solution immediately turns dark brown. It is stirred for 1 hour at room temperature and then poured into ice-cold saturated ammonium chloride solution and extracted with chloroform. The chloroform extract is washed with water, dried 'it over anhydrous magnesium sulfate, decolorized with activated [charcoal and evaporated at 25 °, whereby 60.7 mg of a solid residue yield. This is chromatographed on a 0.5 4.5 cm column of 1.0 g of neutral aluminum oxide (WOeIm ₅ , activity level II) ^: "Eluting with methylene chloride gives 40 mg of solid in the first 5 ml. This is again on a ; 0.5 χ 5.5 cm column of 1.2 g of aluminum oxide chromatographed. Eluting with 5 ml of 1 ml of benzene-methylene chloride yields 28 mg of solid material, the IR spectrum of which shows only a weak absorption at 5.6 μ . Elute with 10 ml of methylene chloride then gives 8.6 mg of a crystalline compound, F 121 to 122. The IR spectrum of this compound (KBr) shows peaks at 2.92 (NH), 5.56 (β-lactam), 5.62 , 5.79: (phthalimido), 5.82 (ester) and 6.03 u (C = C).

The NMR spectrum shows peaks at 2.25 (4H), 3.87 (IH, d, 2.0 Hz 4.30 (IH, d, 2.0 Hz), 6.23 (3H), 7.70 (2H), 7.88 (3H).

88 -

509820/1161

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The connection has the structure

The systematic name for the ring is

I. 1,5-Diaza-6-S-bicyclo- [4,2, Q] -octan-8-one.

! Reaction of methyl 2- (2 ^t -S-chloro-3 ^I -S-phthalimido-4 ^t -oxo) - ■ azetidinyl-3-methyl-2-butenoate with .1 molar equivalent of J N-bromosuccinimide

j A mixture of 165 mg (0.455 mmol) of the pure trans-methyl- . esters, 81 mg (0.455 mmol) N-bromosuccinimide and 4 mg benzoyl-! peroxide in 12 ml carbon tetrachloride is heated to reflux. Then remove the heat source and replace it with a 100 ¥ att Photoflood lamp No. 2. The stirred reaction mixture is illuminated for 15 minutes. Then you cool

509820/1161

M / 12860

the suspension, filtered, and the filtrate evaporated to a white foam. This is on a 0.5 x 13 cm column of 3.0 g of neutral aluminum oxide (Woelm, activity level II) Ehromatographiert. Eluting with methylene chloride gives 206 mg of a 60-40 mixture of monobromo compounds. The NMR spectrum of this mixture has peaks at 2.17 (4H), 3.73 (IH, d, 2Hz), 4.37 (IH ^ d, '2Hz), 5.27-5.65 (2H, m , "two overlapping AF quartets) _s 6.15 (3H), 7.59 (1.2 H), 7.82 - (1.8 H). '/.,..'..'-

The structures of these compounds are

CH ₂ Br- <CH ₃

Methyl 2- (2'-S-chloro-3 ^! -S-phthalimido-4 »-oxo) -acetidinyl-3-methyl-4-br'om-trans-2-butenoate.

and

Methyl 2- (2 »-S-chloro-3'-S-phthalimido-4'-oxo) -acetidinyl-3-methyl-4-bromo-cis-2-butenoate.

-.90-

5098 207 1161

^11/12860 «

Conversion of the cis- and trans-methyl-2- (2'-S-chloro-3 ^! -S-phthalimido-4'-oxo) -azetidinyl-S-methyl ^ -bromo ^ -butenoate into the cis- and trans -Methyl-2- (2 ^! -S-chloro-3 ^l -S-phthalimido-4 ^I -oxo) -azetidinyl-3-methyl-4-azido-2-butenoate

The 60:40 mixture of 205 mg (0.455 mmol) monobrominated Compounds in 4 ml of chloroform are treated with 79.1 mg (0.50 mmol) of tetramethylguanidinium azide. The solution will be instant light yellow. It is stirred for 4 hours at room temperature and then diluted with chloroform, washed with water, treated with Activated charcoal and dries over anhydrous magnesium- '! sulfate. Evaporation then gives 184 mg of the mixed azides. ι IR spectrum: 4.7, 5.55, 5.62, 5.79 µ. ■ ·!

Nuclear Magnetic Resonance Spectrum: 2.17 (4H), 3.76 (IH, m, overlapping doublets),; 4.40 (IH, d, 2.0 Hz), 5.52 and 5.58 (0.8H-, AB quartet of an i Isomers), 5.87 (1.2H, br), 6.15 (3H), '7.67 (1.2H), 7.87 (1.8H) J

■ The structures of these 'connections are

CO ₂ CH ₃

j methyl 2- (2 "-S-chloro-3'-S-phthalimido-4'-oxo) -azetidinyl-3-J methyl 4-azido-trans-2-butenoate

- 91 -

509 820/1161

Z.35-686?

M / 12860

and

HH _:

N J ^

i ■: .0

Cl CH ₃ . *; ■■■

CO ₂ CH ₃ .

Methyl 2- (2 · -S-chloro-3'-S-phthalimido-4'-oxo) -azetidinyl-3-methyl-1-azido-cis-Z-butenoate.

Conversion of 2-benzyl-6- (l ^T -methoxycarbOnyl-2 ^f -methyl- ^: prop-l'-enyl) -l-oxo-3,6-diaza-4-rS-5-R-bicyclo- [3 , 2.0] -hept-2-en-7-one with 1 molar equivalent of a lithium thioalkoxide in

Hexamethylphosphoric acid triamide (hydrolysis of the oxazoline ester • to the oxazolinic acid and rearrangement to the oxygen analog

of penicillin G "■", - ■ "■ '■: .-- ■

The scheme of the above-mentioned reaction is as follows

.Ph

3 I ₀ /

7 6

N-

O"

1. RS Li / KMPT--

CH ₀ Ph

Hill ¹

COOH

50 9 820/1161

M / 12860 .- "'·. ··' '. · ■ - 3 *

PhCH ₂ CO!

wherein R = 11-C ₃ H ₇ , tert-C, H

: H

) N

HMPT = hexamethyl phosphoric acid triamide

; For the various Experiments I described below, HMPT is removed by distillation under dry nitrogen Lithium hydride or lithium aluminum hydride at 116 to 117 C! and cleaned 19-20 torr. It is about molecular sieves in one ! Piston stored, which is provided with a rubber wall, j and withdrawn with a syringe if necessary. The lithium reagent j is created in HMPT by one of the following procedures (procedure A and procedure B):

! Method A : A round bottom flask is provided with a metal stirrer i and a rubber wall »Two! Syringes are inserted and dry nitrogen is passed through: the flask. The HMPT is introduced and the nitrogen purge is continued for 1 hour. Then you add the desired amount of mercaptan and then at 25 an equivalent amount of n-Butylli thiumlö solution in hexane. The nitrogen purge

509820/1> S1

M / 12860

is continued for 15 minutes and then the flask is closed and stored in the refrigerator, with aliquots of the reagent removed as needed for the reaction Solution is stable for about 1 month. ·

Method B: Lithium hydride is placed in a round bottom flask equipped with a magnetic stirrer and a rubber partition. Purge the flask with nitrogen and then add HMPT. It is then flushed with nitrogen for a further 30 minutes before adding the mercaptan. The resulting mixture is stirred for 1.5 hours at room temperature and then filtered off (under a nitrogen atmosphere '' * in a drying chamber). .. The concentration of the reagent is determined by titrating an aliquot with 1 η HCl and then capping the flask and storing it in the refrigerator until use. - ·

Attempt 1 , '

200 mg (0.64 mmolj oxazoline in 1.5 ml HMPT are under a nitrogen atmosphere and stirring with a magnetic stirrer with 1.3 ml (0.75 mmol) of an Ö, '58 molar solute 'of lithium thio tert-butoxide in Hexamethylphosphoric triamide, prepared by process A. The addition is made at room temperature for 0.5 hours and then the reaction mixture is stirred overnight. Ice-cold water and ether are then added and the pH is adjusted to 2.3 Washed with cold water, dried over anhydrous magnesium sulfate, and evaporated to give a yellow oil. This is dissolved in ether and the solution extracted with ice-cold 5% bicarbonate. The bicarbonate extract is adjusted to pH 3 and extracted with ether. Evaporation of the dried ether extract gives 120 mg of yellow oil that ß-lactam absorption in the IR spectrum at 5-6 μ and in the NMR spectrum at 4.8-5.0 "£ * shows. This NMR spectrum changes after 7 hours at room temperature.

- 94 -

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Attempt 2

62 mg (0.197 mmol) of oxazoline in 1 ml of hexamethylphosphoric acid triamide are flushed with nitrogen for 30 minutes and then 0.35 ml (0.215 mmol) are added dropwise over 2 hours a 0.615 molar solution of lithium thio-tert-butoxide, prepared according to process A, to. After the addition is complete The mixture is stirred for 2 hours and the acidic material is isolated as described in Experiment 1. It's 23 mg of one yellow oil.

Attempt 3

66 mg (0.21 mmol) of oxazoline in 3 ml of hexamethylphosphoric acid triamide are flushed with nitrogen and 0.3 ml of a 0.73 molar solution of nC ^ H ₇ Li, prepared according to method A, is then added dropwise in 2 parts. 0.2 ml is added over 0.5 hour, the mixture is stirred for 1.5 hours and then the remainder is added over 0.5 hour. The reaction mixture is kept at room temperature for 8 hours and then stored in the refrigerator overnight. Isolation yields 24 mg of acidic material.

* ι

! Experiment 4 _/ "|

64 mg (0.20 mmol) of oxazoline are dissolved in 1 ml of hexamethylphosphorus j • acid triamide and degas the solution for 45 minutes with a ! Stream of dry nitrogen. Then give for 35 minutes

j I

j 0.44 ml of a 0.47 molar solution of lithium thio tert-butoxide (prepared according to process B) to. "The stirring is continued for 2 hours! and then the acidic material is isolated. It weighs 15 mg.

Any of the acidic compounds found in experiments 2 through 4 were obtained shows NMR peaks at 2.5; 4.0 (d, 3.5 HZ), 6.18, 7.77 and 8.47.

shows NMR peaks at 2.52 (phenyl), 3.77 (d, 3.5 Hz)

- 95 -

509820/1161

M / 12860

3?

j These are the peaks of a compound of the formula

There

COOH

which can be obtained in crystalline form by an alternative process. Each of the NMR spectra of the acids from experiments 2 to 4 also shows peaks that do not belong to them, the most striking of which are a singlet at -t 6.3 and a singlet at 8.75, corresponding to a proton, alpha ζ a carboxylic acid and methyl groups, which are bound to a quaternary center. . ·

It was found that the crystalline ^ oxazoline carboxylic acid - is inactive as an antibacterial agent at doses of 250 γ / ml. However, all acidic fractions produced by hydrolysis of the \ Methyl esters show significant antibacterial activity. The acid from experiment 3 (the "also the biolgosch -inactive oxazoline carboxylic acid), is in dirnethyl sulfoxide dissolved, chromatographed and compared with penicillin G.

The two substances are made on double 1/2 inch strips.

S and S 589 "blue ribbon" dripped paper. After you overnight in a system that uses 60:15:25 n-butanol, Acetic acid and ¥ water contains were developed ..-.- "■ the strips are air dried and a set is made using Riker penicillinase sprinkles. Then both sets are bioautographed on B. subtilis. It shows ,

50 9820/1161

M / 12860

found that the antibacterial agent in the acid from Experiment '3 has an Rf value practically that of penicillin G is identical and the bioactivity is eliminated by treatment with penicillinase as one would with a compound expected, which has a structure similar to that of penicillin G.

Data on minimum inhibitory concentrations of penicillin G and the acid from Experiment 3 are in the table below listed.

- 97 -

509820 / M61

"M / 128bO
Organism penicillin G control Y / ml Acid from experiment 3 MIC .004 MIC γ / ml D. pneumoniae -. ·
+ - 5 ^ serum *. ■ ... " .004 '. - ^: "; . ■ ". ■" ■ 8 .- "'. St. Pyogenes. .
+ $ 5 serum * ■■ .016. ■■ '. ■' ■ "■ ^: V, '■; ■■ · ■ -β ■'. ·· ... ■ ,. ^s · aureus Smith. .
3? ®Α_ ~ ¹⁰ ~ Verd. ^v ... ^! .06 - =: ■; - ■■ · "■■" · ' : ■■; ·· ■ 16 · '■ - "■ S. aureus Smithy.
at IQ " ⁴ Verd ^. *!" ■. ··, .- .. ·.
+ 50% ^ erum 125 '■' ■ ': J ^: ■ ·'
>i25;.,; . \ . ^ v- ■:. & ■. ■■ · .:, .j- S. aureus BX-1.633-2 V - ·.
at 10 ~ ³ comp.
S. aureus BX-1633-2
at ΙΟ "· ² comp. 125; ·, j-,
I ... . ;; .: 125 ■ / '..- ■ S. aureus meth.
Resist, bei'.ior? Verd . "' ⁶ ^ ■■■■ - - ■■" ■. ^.: V / ■■ ·. "'32 - ·. ''. ·. S. aureus at 10 Verd k> I25 \; "'. ■; ■':, S. aureus at "10" '^. Verd ■ 0.13; . ■ /.; . ■ ·· ■ " ^: > ¹² 5 ;; - ^:: " ■ · ^; SaI; enteritidis
. 10 ~ ⁴ Verd. """.'··,; 32 y; r \ _% ■ · ■ '., ■ ^ ^: · 'Κ ·. 63 ■. ·. ··· ■ ·:; · .- "■ E. coil Juhl-'bei / iÖ
Verd. ... ■. """""■ 125. '■■■;_' ■ . "; ■■■ - ■■■. E. coli. "iÖ" ⁴ "" .Vei? d. ■ · ....'-- -4
K. pneumoniae at 10 - ■
Verd.
_t - j ' ¹ V, - ^: ■; "■■:»' ■■■
. ■ ■ * ·. Pr. Mirabilis at 1Ö ~.
Verd. _. · '■ "'': > i25 '': ■ .-.:; \ ^% ■; ■ '■'. .... ■ ς.ν; - 63,; · ■. ·;. ■■ Pr. Morganii at ... 10 " ⁴ V "ν; · ^: Λ.". '63' Γ - · ■■ ^; ·

; ^ Verd.J ·· ',. ■■■. Medium "" 98 - ■

Verdi;

- ·. - ■ - - Ps. Aeruginosa in J.0

Ser. marcescens at .10 "" - ^> 125 '' ·. "..- ■ / ■: · ■ .- /. \ ..: ..": ·· 63 · Verd. <., .- ..,. ···. ■■ - '"'·""; ■ '.":'. ■ · '.' ■

5 0 9820/1161

M / 12 860

Because of its antibacterial spectrum, its sensitivity versus penicillinase and its .NMR spectrum becomes the antibacterial agent that, during hydrolysis of the oxazoline ester is assigned the following structure

• Ö -Ή HH

, 11 I Ξ - C-

_Me

| o .5

Me

Oxapenicillin G.

2,2-Dimethyl-3-R-carboxy-7-S-phenylacetamido-1-oxa-4-aza-5-R-bicyclo- [3,2,0] -heptan-7-one.

Conversion of 2-benzyl-6- (l ^! -Methoxycarbonyl-2'-methylprop-1'-enyl) -l-oxa-3,6-diaza-4S, 5R-bicyclo- [3,2,0] -hept -2-en-7-one with two equivalents of a lithium thioalkoxide in hexamethylphosphoric acid triamide

74 mg (0.236 mmol) of oxazpline are dissolved in 1.5 ml of hexamethylphosphoric acid triamide and degas the solution with dry nitrogen for 0.5 hours. Then give for 0.5 hours 0.83 ml of a 0.612 molar solution (0.508 mmol) lithium thio-tert-butoxide, prepared according to method A, and stir the mixture overnight. The acidic fraction weighs 33 mg and shows antibacterial activity in a platelet test against S. lutea. The NMR spectrum shows ß-lactam protons at 4.0 and 4.7 Hz, phenyl absorption at 2.6, methylene absorption at 6.12, a singlet at 6.2 and methyl peaks at 8.75.

- 99 -

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101

The characteristic peaks of the oxazoline carboxylic acid, 2-benzyl-6- (1 '-carboxy-2 ^f -methyl-prop-'l'-enyl) -I-oxa-3, "6-' diaza-4S, 5R-bicyclo- C3,2,0] -hept-2-en-7-one are not included in the "spectrum. - ■ ".. ·

■ The same result is obtained when trying with

Lithium thio-tert-butoxide, produced according to process B.

These experiments show that hydrolysis of 2-benzyl-6-

■ (l ' ^T -methoxycarbonyl-2' -methylprop-1. '-Enyl) -l-oxa-3,6-diaza-: 4S, 5R-bicyclo- [3,2,0] -hept-2-ene -7-one in the carboxylic acid by lithium thioalkoxide in hexamethy! Phosphorus acid _tri-: amide takes place and that rearrangement in the oxapenicillin under; takes place under the test conditions.

Implementation of the epimeric methyl 2- (2'-chloro ~ 3'-S-amino-4'-oxo) ■ azetidinyl-3-methyl-2-, butenoate with benzyl chloroformate (carbobenzoxychloride)

The crystalline p-toluenesulfonic acid salt of methyl 2- (2 _ ^ - R-chlori-3 ^! - ^ - amino-4 '' - oxo) -azetidinyr-3-methyl-2-butenoate (576 mg, 1.42 mmol) it is dissolved in 6 ml of chloroform suitable for spectroscopy and 1.080 g (5 molar equivalents) of tetramethylguanidinium chloride are added. The mixture is refluxed for 3.5 hours, then cooled, washed with water, dried over anhydrous magnesium sulphate and 297 mg (1. 7 mmoles) of carbobenzoxychloride are added and then a solution of 185 mg (1.83 mmoles ) Triethylamine in 5 ml of dry chloroform. After 45 minutes of stirring at room temperature, the reaction is stopped by adding water. The organic layer is dried and evaporated to a residue of 353 mg. This is chromatographed on silica gel and the column is eluted with 1: 1 petroleum ether (30 to 60 °) - ethyl acetate. Man

-100 - · ■

5 0 9820/1161

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491

collect seventeen 13 ml fractions. Fractions 2 to 4 result "J 108 mg of a compound of the structure

PhCH ₂ O

il

C-

H ₁ H Nfcf

Cl

CO ₂ CH ₃

The NMR spectrum of this compound shows peaks at 2.67 (5H), 4.20 (IH, d, 2Hz), 4.87 (2H) ,. 5.35 (IH, d, 2Hz) ,. 6.27 (3H), 7.73 (3H), 8.03 (3H). ■

Fractions 12 through 17 make 54 mg of one. Connection of structure

· '. HU

HMIl

IMiH

CO ₂ CH ₅

j This fused ß-lactam oxazolidone has a molecular peak j at m / e 240 in its mass spectrum. The IR spectrum shows Peaks at 3.0, 5.56, 5.62, 5.78, 6.1Ou. The NMR spectrum j shows peaks at 2.87 (IH, br), 3.92 (IH, d, 4Hz), 5.08 (IH, d, 4Hz), 6.20 (3H), 7.70 (3H), 8.02 (3H).

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Conversion of 2- (2 '- ^ ehlor - ^' - S-amino-4'-oxo) -azetidinyl-3-methyl-2-butenoic acid with carbobenzoxychloride; v

^H - '■ · - 0, HH. ■ egg

r Cl O WM.- Cl

- COOH

COOH

557 mg '(1.5T mmol ")' p-toluenesulfonic acid salt of anhydro-6-aminopenicillin is chlorinated in methylene chloride at" 0 '. Then the solvent is removed at 20 ° under reduced pressure. The residue is dissolved in a mixture of 4 ml of water and 5 ml of acetone and this solution is kept at 0 for 3 hours in order to carry out hydrolysis of the acid chloride. Then 705 mg (8.4 mmol) of sodium bicarbonate are added and then a solution of 660 mg (1.5 * 1 mmol) of carbobenzoxychloride in 5 ml of dry acetone is added dropwise at 0. The reaction mixture is stirred for 1 hour and the product is isolated by dilution with water, extraction with ether, drying over anhydrous magnesium sulfate and evaporation. The IR spectrum shows peaks at 3.0, 5.60, 5.62, 5.82, _7, 5.88 ^ '. · ■

This experiment shows that the amino acid acylates successfully can be.

- 102- -

50 9 82 0Λ1161

M / 12 860

40V

Chlorination of anhydro-a-phenoxyethylpenicillin

The anhydropenicillin (500 mg of an 80:20 L to D mixture of side chain diastereoisomers) is chlorinated in methylene chloride for 15 to 20 seconds at room temperature Use of a strong stream of chlorine. The solvent is then removed at 20 °. The resulting product is a Mixture of isomers of the formula

Cl-

-0 -

H 0

I Il

C-C

I.
CH ₃

HHH

I = r Cl

-N-

COCl

Its NMR spectrum shows peaks at 2.3 (IH, d, 9Hz), 2.75 (2H, d, 9Hz), 3.13 (2H, d, 9Hz), 3.92 (IH, m), 4.45 (IH, m), 5.28 (IH, m), 7.74 (3H), 7.86 (3H), 8.40 (3H, d, 7Hz). The IR spectrum shows peaks at 2.95, "5.55, 5.91

This experiment shows that a 6-acylamino-anhydro-penicillin can be chlorinated with success. ·

- 103 -

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j hydrolysis of anhydro-a-phenoxyethylpenicillin chlorination ^ ι products

j The above trichloro compound is dissolved in ml of acetone $, I the solution is cooled to 0 and added dropwise with stirring a solution of i 284 mg sodium bicarbonate in 5 ml of ice-cold ■; Water too. Some oily material precipitates out during this addition and therefore 15 ml of acetone is added when the addition is complete. The resulting pale yellow solution is stirred. 2 hours at 0 ° and then diluted with 100 ml of cold water. j Extraction with methylene chloride, drying over anhydrous ¹ magnesium sulfate and evaporation gives a pale yellow oil. 'The following structure is assigned to this material

0 - · C - C - N * i

i The IR spectrum shows peaks at .2.9 / 5.58, 5.85 and one dissolves

j the compound in methylene chloride and extracted the solution

ι with 5 % sodium bicarbonate. The aqueous extract will

^! acidified again to pH 2.5 and again with methylene chloride

j extracted. Evaporation of the dried methylene chloride extract

^: gives material with high antibacterial activity against

ι S. lutea, to which the following structure is assigned

-, 104 '- ■

BOiSZO / ti SI '

M / 12 860

O -

CH.

COOH

Treatment of this material with diazomethane yields a methyl ester which peaks at 5.6 u and in the IR spectrum shows a peak at 8.75 in the NMR spectrum, consistent with the presence of a ß-lactam and methyl groups, the are attached to an oxazolidine ring. '...

In a second attempt, the chlorination product of 215 mg (0.62 mmol) of anhydro-a-phenoxyethylpenicillin in 3 ml of ice-cold tetrahydrofuran dissolved and added to the colorless

2 ml of ice-cold water are added to the solution. The resulting The solution is stirred for 2 hours at 0 °, then diluted with ice-cold saturated sodium chloride and extracted with Methylene chloride. Evaporation of the dried methylene chloride extract yields 277 mg of a white foam. The NMR spectrum this dichloric acid shows peaks at 2.30 (IH, br), 2.72 (2H, d, 8Hz), 3.10 (2H, d, 8Hz), 3.83 (IH-, br), 4.4.0 ( IH, br), 5.22 (IH, q), 7.67 (3H)., 7.93 (3H), 8.40 (3H, d).

The IR spectrum shows peaks at 2.96, 5.60, 5.8 vi

- 105 -

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Conversion of 2- (2 "- chloro-3'-S-amino-4 '-oxo) -azetidinyl-3-methyl-2-butenoic acid with phenylacetyl chloride

5? C1.

HH

PhCH ₀ CONHJf

COOH-

COOH

242 mg of p-toluenesulfonic acid salt of anhydro-δ-amino-penicillin is chlorinated "and hydrolyzed the product, as" already described, in aqueous acetone. To the resulting solution 275 mg of sodium bicarbonate are added in parts at 0 ° (volume 20 ml) and then a solution of 152 mg of phenylacetyl chloride in 5: ml of acetone at once. The mixture is stirred for 30 minutes and then diluted with water and extracted with Methylene chloride. This extract is discarded. One poses the aqueous phase. to pH 3 and extracted with methylene chloride. This extract is dried over anhydrous magnesium sulphate and evaporated to give the desired Connection results. The IR and NMR spectra are consistent with the assigned structure. ■ ■. ■ - "" "." - "-

- 106 -

5 0 9 8 2 0/1 161

M / 12 860

40 *

Chlorination of anhydrobenzylpenicillin ■ -....

176 mg (0.56 mmol) of anhydropenicillin G are dissolved in 7 ml of methylene chloride and the solution is chlorinated at room temperature for 30 seconds with a strong current of chlorine. The solvent is then removed immediately at 20 under reduced pressure, resulting in a pale yellow foam. The IR spectrum of this. Schaums shows peaks at 2.92, 5.5, 5.95 µ. The NMR spectrum shows that the product is a 4: 1 mixture of ZX 2 '-R ^1-chloro-3 -js-phenylacetamido-4' -oxo) -azetidinyl-3-methyl-2-butenoylchldrid and its 2 ^I -S epimer is. The main isomer has the structure ■ -

HH

. PhCH ₂ CONKi

COCl

! Its NMR spectrum shows peaks at »2.71 (5H), 3.07 (IH, d, 9Hz),

3.97 (IH, d, -4.5 Hz), 4.52 (IH, q, 4.5, 9HZ), 6.37 (2H), 7.78 j (3H), 7.92 (3H ):
; The lesser isomer present has the structure

H H

Cl

-N.

eoci.

Its NMR spectrum "shows peaks at 2.71 (5H), 3.07 (IH, d, 9Hz), 4.12 (IH, d, 2Hz), 5.17 (IH, q, 2.9 Hz), 6.44 (2H), 7.78 (6H).

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Hydrolysis of the anhydrobenzylpenicillin chlorination product

The above mixture of chlorides (from 176 mg anhydropenicillin) is dissolved in 6 ml of acetone and 4 ml of water are added and then an excess of sodium bicarbonate at 0 °. The mixture is stirred for 2.3 hours and then diluted with 50 ml of water. and extracted three times with methylene chloride and once with ether. Acidification of the aqueous phase to pH 2.5, extraction with methylene chloride and evaporation of the dried methylene chloride extract gives 36 mg of an oil, its IR and NMR spectra show that it consists mainly of unhydrolyzed Acid chloride consists.

In a second attempt, 105 mg of anhydropenicillin G are chlorinated and the resulting white foam is dissolved in 3 ml of ice-cold tetrahydrofuran and 1 ml of cold water is added. The clear yellow solution is kept at 0 ° for 2 hours and then poured into a mixture of water and ice and extracted with methylene chloride. Evaporation of the dried methylene chloride extract gives 113 mg (98 %) of a chloric acid, the IR and NMR spectra of which show that it has the following structure:

HH
= χ Cl

COOK

The IR spectrum shows peaks at 3.05, 5.62, 5.90, 5.95 W. Das NMR spectrum shows peaks at 2.72 (5H), 3.10 (IH, d, 8 Hz), 3.93 (IH, d, 4.5Hz), 4.42 (IH, q, 4.5, 8Hz), 6.32 (2H), 7.72 x (3H),

8.01 (3H). _f ■ :

- 108 - -.

50982Q / 116 |

M / 12 860

440

In a third experiment, the chloric acid from 333 mg of anhydropenicillin G in ether is treated with an excess of essential diazomethane. The solution is stirred for 2 hours at room temperature and then evaporated to dryness. The residue is left in the refrigerator overnight and then shaken in methylene chloride with 5% bicarbonate, dried and evaporated. The residue is chromatographed on silica gel. Eluting with 1: 1 petroleum ether ": Ethyl acetate gives 189 mg (51 %, F 110.5 to 111 °) of the methyl ester of the formula

H H

PhCH ₂ CONHi *

and 40 mg (12 %) of the oxazoline

Both compounds are identical to those obtained from the p-toluenesulfonic acid salt of "methyl 2- (2'-R-chloro-3'-S-amino-4'-oxo) -azetidinyl-3-diethyl-2-butenoate receives. The reaction sequence shows that the chlorination in the phenylacetamido and amino side chains are made in the same manner. ■

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Synthesis of 2-benzyl-6- (1'-carboxy-2'-methylprop-l - ^! -Enyl) · l-oxa-3 r 6-diazar-4S, SR-bicyclo- [3,2,0] - hept-.2-.en-7-.on

H H

PhCONHf:

Eat

NaHGO.

eooH -

The chloric acid, prepared from 640 mg anhydropenicillin G, is dissolved in 5% bicarbonate solution. This is extracted with methylene chloride and acidified, then to pH 2. Extraction with methylene chloride and evaporation of the dried extract gives a crystalline residue. "Recrystallization from ethyl acetate gives 320 mg (53 %) of the oxazoline carboxylic acid, melting point 119 to 122 ° (decomposition". ")." The "" 1R spectrum shows a broad peak at 4 Ii and peaks at 5.64, 5.92, 6, Iu, consistent with the zwitterionic structure -.- ^: ".

"CS ₂ Ph-Vv _: .- -. ■ - ■

The NMR spectrum (in D ₂ O) shows peaks at 2.67 (5H), 3

(IH d, .3,8Hz), "k _t 15 (IH, d, 3.8 Hz), 6.25 (2K), ^8.09 (3H),

8.69 (3H). ; - ^;

- 110 -

820/1

M / 12 860

Analysis C

calculated: found: -

Manufacture of anhydrocarbobenzoxypenicillin

C. , 99 H 37 N 33 63 , 26 5, 89 9, 54 64 4, 9,

g (7.84 mmol) of p-toluenesulfonic acid salt of anhydro-6-aminopenicillin it is dissolved in 20 ml of methylene chloride, the solution is cooled to 0 ° and treated at the same time with 820 mg of pyridine in 15 ml of methylene chloride and 1.400 g (8.1 mmol) of carbobenzoxychloride in 15 ml of methylene chloride. The addition of the two reagents is complete after 10 minutes and after more The mixture is washed for 10 minutes with ice-cold 1 η hydrochloric acid, ice-cold saturated sodium chloride, dry and evaporate to give 2.740 g of a white foam. This is made from a mixture of ether and petroleum ether crystallized, resulting in 2.33 g (89.5%) anhydrocarbobenzoxypenicillin with the following formula:

PhCH ₂ OCONH ^

j The NMR spectrum shows peaks at 2.65 (5H), 4.17 (IH), 4.45 ! (IH), 4.85 (2H), 7.82 (3H), 7.93 (3H).

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Her position of anhydro-2,2 ', 2 ^f ' -trichloräthoxypenicillin

The 2.396 g (6.47 mmol) of p-toluenesulfonic acid salt of anhydro, 6-aminopenicillin is dissolved in 25 ml of methylene chloride., Cools the solution and treated simultaneously with 0.633 g (8 mmol) of pyridine in 10 ml of methylene chloride and 1.437 g (6.9 mmol). 2,2 ', 2 "-Trichloräthylchlorformiat in 10 ml of methylene chloride. When the addition is complete, thin-layer chromatography gives the Reaction mixture a single spot (and a spot on the Origin, which corresponds to pyridinium chloride). One stirs them Mixture for another 15 minutes, then washed with cold 1 η HCl, dries and evaporates. The IR and NMR spectra of the resulting white foam are consistent with the structure

Ο Η

ΟΙ

Gl - C - ■■ CH ₀ OC -

H H

■ ■ * ■■ *

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509820 / Π 61

M / 12 860

Conversion of anhydro-2,2 ', 2' '- trichloroethoxypenicillin to Benz.iiydryl-2- (2 '-R-chloro-3' -S-trichlorethoxycarbonylamino-4 ' -oxo) -azetidinyl-3-methyl-2-ldutenoate

^{H H}

· Ο

?. 5 egg

COCl

H H Cl ■ ·

CCl, CH, pCONH-f

■ • ■

■ N

CO ₂ H

i Cl

CCl ₃ CH ₂ OCONH-J

■> ·

H ₂ O-THP

- ^

B ^:

CO ₂ CHPh ₂

- 113 τ

509820/1161

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(A) The 453 mg (1.245 mMor) "Aiih.ydropeniciilin are as usual chlorinated, resulting in "a white foam." The NMR spectrum. this foam shows peaks at 3.35 (IH, d, 10 Hz), 3.83 (IH, d, 4.0, 10 Hz), 5.21 (2H), 7.71 (3H), 7.81 (3H).

(B) At 0 ° for 2 hours in 10 ml of tetrahydrofuran and 1 ml. Hydrolyzed water. The resulting acid crystallizes from chloroform. The NMR spectrum shows peaks at 3.17 (IH, d, 8Hz), 3.82 (IH, d, 4Hz), 4.56 (IH, q, 4.8 Hz), 5.20 (2H), 7, ^ 7 (3H), 7.93 "(3H).

(C) The acid is dissolved in 40 ml of hot benzene and added

363 mg (1.87 mmol) of diphenyldiazomethane are added. The resulting solution is refluxed for 40 minutes and then evaporated to dryness. The dark red oil is chromatographed on silica gel. Eluting with graded mixtures of petroleum ether and ethyl acetate yields 480 mg (70 %) of the .benz-, hydrylester. Its NMR spectrum shows the peak "s" at 2.68 (10H), 3.06 (IH), 3.51 (IH, d, 9 Hz), 4.04 (lH, d, 4.5 Hz), ' 4.67 (IH, q, 4.5, 9Hz), 5.23 (2H), 7.72 (3H), 7.98 (3H).

Equilibration of the R- and S-benzhydryl-2- (2-chloro ^{J-3'-S-trichloräthoxycarbonylamino-4'-oxo,)} - azetidinyl-3-methyl-2-butenoates ·

The 181 mg (0.317 mmol) of benzhydryl esters and 240 mg (5 mol equivalents) of tetramethylguanidinium chloride are refluxed in 5 ml of chloroform for 5 hours. The cooled reaction mixture is then diluted with methylene chloride, washed with saturated sodium chloride solution, dried and evaporated. The resulting white foam turns out to be a 60:40 mixture of the starting material and its 2 ^I S epimers of the formula

- 11 4 -

9820/1161

M / 12 860

CO ₂ CHPh ₂ ,

The NMR spectrum of this compound shows peaks at 2.68 (10H), 3.06 (IH), 4.30 (IH, d, 2Hz), 4.95 "(IH, d," 2Hz), 5.27 (2H), 7.72 (3H), 7.98 (3H).

In a second experiment, 4.851 g (8.65 mmol) of the 2'-R isomer and 7.9 g (6 molar equivalents) of tetramethylguanidinium chloride are refluxed in 20 ml of spectroscopic chloroform for 4 hours. The product is a 70:30 mix of 2 ^! -S: 2'-R isomers. This means that these conditions, where there is a higher concentration of reactants, produce a significant improvement in the proportion of the 2'-S-isomer.

Tetramethylguanidinium formate

Commercially available 98% formic acid is converted from anhydrous Copper sulfate distilled at 46 ° and 113 Torr. That The distillate is distilled again from anhydrous copper sulfate at 41 ° and 105 torr, with anhydrous formic acid results.

- 115 -

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22 g (0.19 mol) of tetramethylguanidine are dissolved in 100 ml of anhydrous Ether and add to this solution at 0 ° a solution of 7.1 ml (0.198 mol) of anhydrous formic acid in 40 ml of anhydrous ether. The resulting salt crystallizes. It is collected, washed with ether, dried in a high vacuum and recrystallized from chloroform-ether.

The salt is extreme, hygroscopic and becomes oily, after it '' Exposed to atmospheric moisture for 3 minutes. That oily material gives unsatisfactory results in subsequent experiments Results.

If the formic acid is not "anhydrous" before the salt formation if you do, you get this as an oil that is unsatisfactory Delivers results. This salt has the formula

OrC-H "

Il '
Me ^ NC-NMe

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Conversion of benzhydryl 2- (2'-chloro'-S-trichloroethoxy- "carbonylamino-4'-oxo) -azetidinyl-3-methyl-2-butenoate with tetramethylguanidinium formate. Production of 2-trichloroethoxy-6- (1 ' -benzhydryloxycarbonyl-2'-methylprop-1'-enyl) 1-oxa-3,6-diaza-4-S, 5-R-bicyclo- [3,2,0] -hept-2-en-7-one

A 2: 1 mixture of 953 mg (1.7 mmol) of 2'-S: 2 _'T R Benzhydrylestern and 4,992 g (30.5 mmol) of tetramethylguanidinium formate heating them in 12 ml of chloroform spectroscopy suitable to reflux. The reaction is complete after 1.5 hours and the mixture is therefore cooled, washed successively with water and saturated sodium chloride, dried and evaporated. The residue is chromatographed over 30 g of silica gel. It is eluted with 100 ml of 90/10, 80/10 and 70/30 petroleum ether-ethyl acetate and 30 ml fractions are collected. Fractions 2 to 5 are combined, 142 mg (24 %) of a compound of the following formula being obtained

ei .- ■ ···
ι ·.

OCH ₀ C-Cl. ν

I " L

■ N

IHlH

..NL

Ph

The IR spectrum of this oxazoline shows peaks at 5.61, 5.76, 5.81, 6.07 μ

- 117 -

609820/1161

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The NMR spectrum shows peaks at 2.68. (10H), 3.08 (s, IH), 3.99 (IH, d, 3.5Hz), 4.86 (IH, d, 3.5Hz), 5.15 (IH, d, 12Hz), 5.27 (IH, d, .12Hz, the methylene protons of the trichloroethyl group are non-equivalent), 7.73 (3H), 8.06 (3H).

The mass spectrum shows chlorine multiplets for the presence of 3 chlorine atoms are characteristic, including a multiplet at m / e 522, the molecular ion.

'Chlorination of anhydrocarbobenzoxypenicillin ν

; 470 mg (1.47 mmol) anhydropenicillin are 15 seconds in

; Chlorinated methylene chloride. After another 30 seconds, will

: the solvent removed, leaving a white foam,

which consists of a 77:33 mixture of dichlorides, where

; ' ^{the 2 f} -R-chlorine configuration predominates in the compounds.

It has the following structure

^{0 H}

PhCHJD .-- C -Nfs

1 · Ό

■. COCl

The NMR spectrum of this compound shows peaks at 2 ^ 69 (5H), 3.14 (IH, d, 9Hz), 3.93 (IH, 'd, 4Hz), 4.75 (IH, q, 4.9Hz), 4.83 (2H), 7.80 (3H), 7.90 (3H).

- 118 -

SO 982 0/1161

M / 12 860

The isomer present to a lesser extent has the structure

HH

PhCH ₂ O - C - N?

COCl

The NMR spectrum of this compound shows peaks at 2.69 (5H), 3.14 (IH, d, 9Hz), 4.10 (IH, d, 2Hz), 4.84 (2H), 5.17 (IH, d, 2Hz), 7.80 (6H). ·

Hydrolysis of the anhydrocarbobenzoxypenicillin chlorination product.

j The dichloride (a 77:23 mixture of 470 mg 2'-R: 2'-S Anhydropenicillin) is dissolved in 10 ml of cold tetrahydrofuran ! and add 1 ml of ice-cold water. The mixture is left j 3 hours at 0 and then adds them to ice-cold saturated sodium chloride solution. Extraction with methylene chloride and ! Evaporation of the dried methylene chloride extract gives 535 mg of the chloric acids. The main isomer present has the structure

O H

H H

PhCH ₂ O -C-

COOH

119 -

50 9 820/11 61

Μ / Ί. -2 860

f <f

The NMR spectrum of this compound has peaks at 2.68, 3.63 (IH, d, 10Hz), 3.87 (IH, d, 4Hz), 4.55 (IH, d, 4.10Hz), 4.83 (2H), 7.72 (3H), 7.98 (3H).

Reaction of 2- (2 ^t -R-chloro-3 ^t -S-benzyloxycarbonylamino-4 ^I -oxo) -azetidinyl-3-methyl ~ 2-butenoic acid with sodium bicarbonate. Preparation of the oxygen analog of a 5-epianhydropenicillin

Three chloric acid from 470 mg of anhydropenicillin in 5 ml of acetone is added to a solution of 247 mg (2.94 mmol) of sodium bicarbonate in 10 ml of water. Crystallization begins after 1 minute. The mixture is stirred for 15 minutes and then diluted with 20 ml of saturated sodium chloride solution and extracted twice with 10 ml of methylene chloride each time and once with 10 ml of ether. The combined organic extracts are dried over anhydrous magnesium sulfate and evaporated. The crystalline residue weighs 216 mg. Chromatography on silica gel gives 185 mg of material which is recrystallized to give a compound with m.p. 170 to 172. ^ ⁵

(decomposed),
Has

-64.5 ° (c 0.45, CHGl ₃ ), the following formula

■ PhCH ₂ O

-120 ~

509820/1161

M / 12 860

Anhydrocarbobenzoxypenicillin has [a] _D + 259 ° (c 1, CHCl,).

The 5-epoxy anhydropenicillin has IR absorptions at 2.98, 5.55, 5.65, 5.88 and 6.51 u The NMR spectrum has peaks at 2.44 (6h), 4.14 (IH, d, 1.8 Hz), 4.82 (2H), 4.98 (IH, q, 1.8, 8Hz), 7.82 (3H), 7.88 (3H). The mass spectrum shows the molecular ion at 316.

analysis

"calculated;:
found:

^J 5

60.75

60.67

5.10

5.20

8.86

8.99

Manufacture of 6-phthalimido-oxapenici.llin

606 mg of Änh.ydro-6-plrfch.alimidopenicillin are chlorinated in methylene chloride for 30 seconds with a strong stream of chlorine and the solvent is then removed immediately. A 70:30 mixture of 2 ^l -S: 2'-R 2 is obtained - (2'-Chloro-3'-S phthalimido-4 ^f -oxo) -azetidinyl-3-methyl-2-butenoyl chlorides.This mixture is hydrolyzed for 4 hours at 0 in a mixture of 25 ml of tetrahydrofuran and 2 ml of water. The resulting acid is a 77:23 mixture The major isomer present has the structure

• COOH

- 121 -

509820/1161

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NMR spectrum has peaks at 2.12 (4H), 3.62 (IH, d, 2Hz), _4.38 (IH, d, 2Hz), 7.84 (3H), 8.11 (3H) . The isomer that occurs to a lesser extent, with the 2. ' -R configuration was made

j already described. ', ...,. . . . .

! The trans-acid is crystallized from 1: 1 ethyl acetate-ligroin, i P 164-166 ° (decomp.) (Sealed capillary tube). The crystalline trans-acid in methylene chloride is shaken for 5 minutes; 5% bicarbonate. The aqueous phase is then separated off. ^: at pH 4.5 "to 5.0 acidified and with methylene chloride

extracted. It is then acidified to pH 2.5 and extracted I twice with Me-Öylenehl'orid and twice with chloroform. A- ; evaporate the combined dried organic extracts yields 30 mg of material that has antibacterial activity against

S. lutea shows at 1/10 the dose of phthalimidopenicillin.

The NMR spectrum shows a peak at 8.73 for methyl groups ! on an oxazölidine ring and ß-lactam absorption at 4.6. ■ The active connection has the following structure

- 122 - ■ -. "

5 0 9: 82: 0! /.

2356362

M / 12 860

Preparation of benzhydryl 2- (2 '-S-chloro-3' -S-phthalimido-4 ¹ -oxo) -azetidinyl-3-methyl-2-butenoate

526 mg (1.515 mmol) of the crystalline 'trans-chloric acid are dissolved in 12 ml of boiling ethyl acetate, the solution is then cooled to room temperature and treated with 330 mg (a 10 % excess) of diphenyldiazomethane in 18 ml of ethyl acetate. The formation of nitrogen begins immediately and is complete after 10 minutes. The mixture is then refluxed for 1 hour and evaporated to dryness. The residue is chromatographed on 30 g of silica gel. Eluting with petroleum ether-ethyl acetate (70-30) removes some unreacted diphenyldiazomethane. Continued elution with petroleum ether-ethyl acetate (1: 1) gives 772 mg (99 %) of the compound of the formula

0,

H ■ H

Ph

The MR spectrum shows peaks at 2.23 (4H, d), 2.67 (1OH, m), 3.00 (IH, s), 3.87 (IH, d, 1.8Hz), 4.47 (IH, d, 1.8 Hz), 7.68 (3H), 7.90 (3H).

-123 -

5 0 9 8 2 0/1161

M / 12 860

Conversion of benzhydryl 2- (2'-S-chloro-3'-S-phthalimido-4'-oxo) -azetidinyl-3-methyl-2-butenoate with tetramethylguanidinium formate. Preparation of Benzhydryl-2- (2'-R-formyloxy-3'-S.phthalimido-4'-oxo) -azetidinyl-S-methyl ^ - butenoat "- ■ - -

The 150 mg (0.292 mmol) benzhydryl ester and 260 mg (1.61 mmol, 5.5 molar equivalents) of tetramethylguanidinium formate is heated Reflux in chloroform suitable for spectroscopy for 17 hours. The solution is washed successively with water, saturated sodium chloride and water and dried over anhydrous Magnesium sulfate. After removing the solvent, the residue is chromatographed over 18 g of silica gel. Elute with graded mixtures of "petroleum ether-ethyl acetate gives 54 mg of a crystalline compound of the following structure

! ι

IHO-C-H:

• Ph

\ The NMR spectrum of this compound has; Peaks at -0.74 (IH), ί 2.19 (4H, d), 3.02 (IH, s), 2.67 (1OH, m), 3.92 (IH, d, 3.8 Hz ),

\ 4.59 (IH, d, 3.8Hz), 7.75 (3H), 8.11 (3H).

-124 -

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Curtius rearrangement of the anhydrobenzylpenicillin chlorination product

206 mg Amhydropenicillin G chlorinated to 20 seconds at -6 ⁰ C in methylene chloride. The solution is kept at this temperature for 2 minutes and then evaporated to dryness. The residue is treated in 10 ml of chloroform at -6 ° with 120 mg (1.2 molar equivalents) of tetramethylguanidinium azide in 5 ml of chloroform. The resulting solution is kept at -6 ° for 1 hour and its IR spectrum is then determined. It shows peaks at 3.0, 4.67, 5.60, 5.95 µ, as is the case for the formation of a compound of the formula

Il

PhCH ₂ C

JC.

• is to be expected. j

The acid azide is isolated by extracting the chloroform solution with water and sodium chloride solution, drying and removing the solvent. It is then dissolved in 10 ml of methylene chloride and then the solution is heated to reflux. After 15 minutes the IR spectrum shows a new peak at 4.43 u> which is about twice as intense as the peak at 4.67 u. After a further 15 minutes the peak at 4.43 τα is about six times as intense as that Peak at 4.67 U. After a total of 2 hours of refluxing, the reaction appears complete and the IR spectrum shows peaks at 3.12, 4.42, 5.58, 5.98 u.

- 125 - ■

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M / 12860

Evaporation gives a compound of the formula

. H ^! H

PhCH ₂ CONHf ^

This isocyanate in 20 ml of tetrahydrofuran is added over 40 minutes to 1: 1 aqueous tetrahydrofuran which contains 0.65 ml of 1 HCl. The mixture is stirred 80 minutes after the addition has ended and then diluted with 200 ml of water, saturated with sodium chloride and extracted exhaustively with methylene chloride. The methylene chloride extract is dried over anhydrous magnesium sulfate and evaporated to give 139 mg of a white foam. The NMR spectrum of this foam shows β-lactam peaks in the range 3.9 to 4.7 ^, the absence of peaks in the range of allylic methyl groups and several peaks in the range 8.5 to 9 T ". Chromatography on silica gel gives two compounds. One, a crystalline material, is determined to be isobutyramide [(CH ^) ₂ CHCONH ₂ ], the other is a 2: 1 mixture of epimers of the formula.

• PhCH ₂ CONH

126 -

509820/1161

PhCH ₂ CONH-I

-N.

M / 12860 the formula H H The cis isomer ■

CH,

\ has NMR peaks at 2.63 (5H), 3.5 (IH, d, 8Hz) ,. 3.94 (IH, d, "5HZ), • 4.32 (IH, q, 8Hz), 6.28 (2H), 6.83 (IH, m), 8.75 (6H, m). j

The trans isomer of the formula

H ]

PhCH ₂ CONH - ^ - ^

CH ₅ ;

CH O 5

has NMR peaks at 2.63 (5H), 3.5 (IH, d, 8Hz), 4.07 (IH, d, 2Hz;), 5.48 (IH, q, 2.8Hz), 6.33 (2H), 6.33 (IH, m), 8.75 (6H, m).

The mass spectrum of the mixture of isomers shows a j M-I peak at 306 and 308 and a peak at 272, respectively the loss of chlorine from the molecular ion.

- 127 -

5 0 9 8 2 0/1161

M / 12860

As previously described, the present invention encompasses a variety of methods including those will be described in detail below. To avoid unnecessary repetitions, make sure that the in the following equations are often based on the substituents of the formula

1 ■ "- ·

R - N- ■■■■■:

2 1

Reference is made, in which often R is hydrogen and R Mean acyl. Acyl is a group of the formulas

Ar -

It

CHC -;

Ar-X-

CR ⁶

II.

■ "Il

Ar -C-

Ar

Il

- R

1Ö

ο ·

It

u-,

Ar-NH-

- OH

Z- 0.
ρ ι. Ii

z-c-c-;

R - NH -

Il

Pm.

Il

0 - C

Understood,

- 128 -

509820/1181

M / 12860 "

12th

wherein R is 2,2,2-trichloroethyl or benzyl;

4th
. R is hydrogen, amino, carbobenzoxyamino, phenyl, fluorine,.

Chlorine, bromine, iodine, hydroxy, lower alkanoyloxy or lower alkoxy;

X oxygen or sulfur; . E.g.

R ⁹ and R, which can be the same or different, are hydrogen, phenyl, benzyl, phenyl or lower alkyl; R ⁷ lower alkyl;

R ^ and R-, which can be the same or different, lower alkyl, Lower alkylthio, benzylthio, cyclohexyl, cyclopentyl, Cycloheptyl, benzyl, phenethyl, phenylpropyl, furyl, T-hienyl, Naphthyl or Ar; '

R lower alkylamino, di- (lower) -alkylamino, cycloalkylamino. With 3 to 7 carbon atoms including, allylamino, diallylamino, phenyl- (lower) -alkylamino, morpholino, lower- (alkyl) -morpholino, di- (lower) -alkylmorpholino, Morpholino- (lower) -alkylamino, pyrrolidino, lower-alkylpyrrolidino, di- (lower) -alkylpyrrolidino, N, N-hexamethyleneimines, piperidino., Lower-alkylpiperidino, di- (lower) -alkylpiperidino, 1,2,5,6-tetrahydropyridino, N - (lower) -alkyl-, piperazino, N-phenylpiperazino, N- (lower) -alkyl- (lower) -. alkylpiperazino, N- (lower) -alkyl-di- (lower) -alkylpiperazino, furfurylamino, tetrahydrofurfurylamino, N- (lower) -alkyl-N-furfurylamino, ₄ N-alkyl-N-anilino or lower alkoxyanilino;

I 2 "5 '

Z, Z and Z, which can be the same or different, lower alkyl or Ar; . "

R lower alkyl, lower cycloalkyl, naphthyl, benzyl,

Phenethyl or. "."

¹ · 1!

Ar-C-

- 129 509820/1161

M / 12860 ' ⁴ M.

and Ar is a monovalent residue of the formulas ",

¹ I 2 3
wherein R, R and R, which can be the same or different, are hydrogen, chlorine, bromine, iodine, trifluoromethyl, phenyl, lower alkyl or lower alkoxy, provided that only one of the

■ 1 -2 ■ '3 - groups R, R and "R can be phenyl, and in particular hydrogen, hydrogen tosylate, phenylacetyl, phenoxyacetyl, carbobenzoxy, trichloroethoxycarbonyl, α-aminophenylacetyl,

1 2

a-Carbobenzoyloxyaminophenylacetyl and when R and R in combination, with the nitrogen atom to which they are bound, be taken, mean phthalimides. Same structures are also referred to as acyl- in some of the following equations. group ■ -.; .._ '\:; .

"..- ■ - ■ - ■ ■■ -. O

^: ■■■■ "" 1- «■» "
shown. ^ · '

In some of the following equations "are esters of the carboxyl group as· .

! I

- C - OR '

shown; in such cases R is lower alkyl and preferably methyl or tert-butoxy, trichloroethyl, benzhydryl or benzyl.

Taking into account the above definitions, the invention preferred procedures summarized as follows:

-130 -

509820/1161

A. Process for the preparation of a compound of the formula

R ² Cl

R ¹ - N - CH CH

I I .C N - C ο

o ^N c - ei

1 2

where R is hydrogen or acyl and R is water

Substance or in connection with R and the nitrogen atom, to which they are bound, phthalimido means there-> ·. characterized in that one is a compound of the formula

1 'ι ^
RN-CH-CH C = O

i I

£ - NC = C (CH ₃ ) ₂

1 2

in which R and R have the above meanings, chlorinated [wherein preferably the chlorinating agent is chlorine, sulfuryl chloride or a complex of chlorine and pyridine hydrochloride is]; . >

B. Process for the preparation of a compound of the formula

R ² 'Cl

R-N-CH-CH

G N -C =

O ^ I ₀

C ^ - OR

1 ?

where R is hydrogen or acyl, R is hydrogen or, in combination with R and the nitrogen atom to which they are attached, is phthalimido and R is lower alkyl, trichloroethyl, benzhydryl or benzyl, there-

- 131 - ' 509820/1161

M / 12860

characterized in that one is a compound of the formula

R ² egg

1 '"■ I
RN - CH CH

I I

CN - C - C (CH _x )

⁰ I / O ^

: _; C ^ V Cl / - ._ "; ■.

1 2
wherein R, R and R have the above meanings, with

mixed with an alcohol of the formula ROH;

C. Process for the preparation of a compound of the formula

R ² Cl

R ¹ - "N-CH-CH
II

. ■■ - ■ c-oH; . ' -

1 2

where R is hydrogen or acyl and R is water

1 "■■ - ■■ - ■■ substance or in connection with R and the nitrogen atom,

to which they are bound, phthalimide © means thereby characterized in that one is a compound of the formula

R ² ' CH Cl ν - C (CH I.
R ¹ - I.
N - I. ι ■■■■■■ -
CH C = j - Cl N - C.

1 2

where R and R have the above meanings, mixed with water; - ■ '\ _:

5 0 9 8 2 071161

M / 12860

D. Process for the preparation of a compound of the formula

• R ¹

CH-CH

, C-N C = Cl

o ^ Lo ^

C ^ ~ OR

where R is benzyl or phenoxymethyl and R is lower alkyl (and preferably methyl or tert-butyl), trichloroethyl, Benzhydryl or benzyl mean, thereby marked -t that one is a compound of the formula

0 Cl , ■ ■

ι Il I

R-C-NH-CH CH

■ I I ·

C - N - C = .C (CH ^ _O. 0

1
wherein R and R have the above meanings, mixed with aluminum oxide or silica gel;

E. Process for the preparation of a compound of the formula

, 4

R ² N, ■

1 "R ¹ - N - CH CH

II '/ ν = C (CHj ₂

^ - OR

- 133 -

509820/1161

2356B62

Μ / 12860

λ ■ -.-. · ... "■ - ■ - · ■ · ■ - ■ - ■". ■ ρ ■ "■■""■■'where R is hydrogen or acyl, R is hydrogen

1
or in connection with R and the nitrogen atom to which they are attached, 'denotes phthalimido and R denotes lower alkyl (and preferably methyl or tert-butyl), trichloroethyl, benzhydryl or benzyl, characterized in that a compound of the formula

R ²
R ¹ -N-CH

1 2
wherein R, R and R have the above meanings, can react with azide ions;

F. Process for making a compound of the formula

R ² Cl

ι f '■■■■■

R - N- CH T-CH _Λ .CH ₂ Br

C ^ ~ OR ■■■■. .

1 ^: 2

where R is hydrogen or acyl, R is hydrogen

1
or in connection with R and the nitrogen atom to which they are attached, Phthalimido 'means, and R means lower alkyl (and preferably methyl or tert * -butyl), trichloroethyl, benzhydryl or benzyl, characterized in that a compound of the formula

- 134 -

509820/11

M / 12860

Ν -C = C (CH) ₅ ,

T 2
wherein R, R and R are as "meanings" above, reacted with about 1 mole of an allyl brominating agent (where preferably the allyl brominating agent is N-bromosuccinimide and a catalytic amount of an organic peroxide is present);

G. Procedure for. Preparation of a compound of the formula

R ² Cl

1 'I.

R ¹ - N - CH CH Q _{11 N}

I I · / 2 5

•. C-OR

1 2

where R is hydrogen or acyl, R is hydrogen or in conjunction with R and the nitrogen atom to which

they are bonded, phthalimido and R is lower alkyl (and preferably methyl "or tert-butyl), trichloroethyl, Benzhy.dryl or benzyl; characterized in that a compound of the formula

- 135 509820/1161

M / 12860 13Y

R ² Ci

"1 I '■ r ¹ - ν -.CH --—CH

ι I / -ä.

G ^ - OR

1 2

wherein R, R and R have the above meanings, with about 1 mole of azide ion (preferably wherein the azide ion source Is tetramethylguanidinium azide);

H. Process for the preparation of a compound of the formula

• R γ>.

R ¹ -N - CH- CH: - _v:

C κ - C =

C ^ r OR '

1 2

where R is hydrogen or acyl, R is hydrogen or in connection with R and »the nitrogen atom to which they are bound, phthalimido and R is lower alkyl, Trichloroethyl, benzhydryl or benzyl means, characterized in that a compound of the formula

R ^x - N - CH-CH, _mH ^^ I fu

0 I ^ "" ¹ X

C ^ - OR:

1 2 "

wherein R, R and R have the above meanings, hydrogenated (preferably in the presence of a platinum oxide catalyst); .. · ■. "■■

- 136 - ■ .-; ■

509820/1161

M / 12860 s 9 * :

I. Procedure for. Preparation of a compound of the formula

0> 0,

1 .

in which R is benzyl or phenoxymethyl, characterized in that a compound of the formula

τ ¹

CH-CH .C-N-C =

o I / O

C "- OR

where R is benzyl or phenoxymethyl and R is lower alkyl (and preferably methyl or tert-butyl), trichloroethyl, Benzhydryl or benzyl, with about 1 mole of a compound of the formula R SLi, wherein R is lower alkyl (and preferably n-propyl or tert-butyl), dissolved in the compound of the formula

O (CH ₃ ) ₂ N -P- N (CH ₃ ) ₂

N (CHj) ₂ -reacted;

- 137 -

503820/1161

Μ / 12860

₄ β * - ;.

J. Process for the preparation of the compound of the formula

OH -C

^N -CHCOOH

characterized in that one uses a decylating agent - a compound of the formula ·,. , /, -. ,

0 / Os

R-C-NH-CH- -CH C

.C - ■ »■ - - CHCOOH

1 - ^:

where R is benzyl or phenoxymethyl, and the deacylating agent is preferably S. lavendulae or E. coli, or in which the deacylation process is carried out by successive addition of trimethylchlorosilane, phosphorus pentachloride and methanol ;. ■■ -; - '.- . ♦.

K. Process for the preparation of a compound of the formula

R - C ■ - NH. - CH

Il ι ^ ⁿ 3

C - N - CHCOOH

1 "
wherein R - C - acyl, characterized in that the compound of the formula

- 138 -

509820/1161

M / 12860

- CH— CH σ ^J

^H 3 C N -CHCOOH

1 'with a carboxylic acid der. Formula R - COOH or his Equivalent as acylating agent for a primary amino group acylated, especially the process in which the Acylating agent the corresponding carboxylic acid chloride, acid bromide and anhydride, the mixed acid anhydride

with; a lower alkyl ester of carbonic acid or a free acid in conjunction with a carbodiimide.

In a further preferred embodiment of this method becomes the acyl group

1 "
R - C - by the formula

shown in which B is an easily removable blocking group and is especially selected from hydrogen chloride acid, carbobenzyloxy, aliphatic ß-diketones and aliphatic ß-diketoesters, for example hydrogen chloride or methyl acetic ester;

- 139 -

509820/1161

M / 12860

L. Process for the preparation of a compound of the formula

0 H,

R ¹ - C - NH - CH - CH "^ CH ₉ i II <v ^ CN C- CH ₃ ,

COOH

0 ^%

1 «■■" - ■■
wherein R - C - is acyl, characterized in that

^ - that one is the compound of the formula

E ₅ N-CH-CH CH ₀

^ i i l ^

^, C__N C - CH,

^ ³

COOH

acylated with a carboxylic acid of the formula R - COOH or its equivalent as acylating agent for a primary amino group, and in particular the process in which the acylating agent combines the corresponding carboxylic acid chloride, acid bromide, acid anhydride, the mixed acid anhydride with a lower alkyl ester of carbonic acid or a free acid with a carbodiimide; . _ _# '- ·

In a further preferred embodiment of this method becomes the acyl group

1 '8

R - C - by the formula.

NHB

- 140 -

50SS20 / 1161

M / 12860.

represented, wherein B is an easily removable blocking group and is in particular selected from Hydrochloric acid, carbobenzyloxy, aliphatic ß-diketones and aliphatic ß-diketoesters, for example Hydrogen chloride or methyl acetic acid ester,

M. Process for the preparation of the compound of the formula

H ₀ N - CH CH CH ₀

COOH

characterized in that a compound of the formula

H
O

ι ι '

i ^x - f! -

Il X

R ¹ - C - NH - CIi -CH CH

COOH

1
where R is benzyl or phenoxymethyl, reacts with a deacylating agent, and the deacylating agent is preferably S-. is lavendulae or E. coli, or the deacylation process is carried out by sequential addition of trimethylchlorosilane, phosphorus pentachloride and methanol;
and

- 141 -

509820/1161

M / 12860

N. Process for the preparation of a compound of the formula

I -. - - '. CH -
I. /
-CH
ι I. -CH ^ R »- I. I.
/ y . a- [ -. I. - I.
COOH C.

1 '2

wherein R is hydrogen or acyl "and R is iiasser-

.- ■■■ - Λ

substance or in connection with R and the nitrogen atom * to which they are bound, means phthalimido, characterized in that -daß'man a compound of the formula

R ² Cl
R ¹ - Ji - CH - ^ - CH

Ü C = C

^ - OR

1 '2
where R. and R is as defined above and R is lower alkyl, trichloroethyl, benzhydryl or benzyl, saponified, preferably with potassium tert-butoxide.

The 4-chloroazetidin-2-ones according to the invention are useful starting materials for the preparation of the class of compounds called secopenicillins "and are described in detail in Belgian patent specification 754 125 (Farmdoc 10051S); they are prepared by reaction with sulfur-containing compounds such as hydrogen sulphide, sodium hydrogen sulphide , Methylnjercaptan and benzyl mercaptan or salts thereof _v (shown below as RS Φ), converted into secopenicillins; andel1 :. ■

- 142 -

50 98 20/1161

M / 12860

ty

Suitable compounds according to the invention are converted into known cephalosporins as follows:

R -

R ² HH

^ Cl

/ CHpX

COOR

^CH 2 ^Y

NH ₂ SH Me ₂ N -C- NMe ₂

CH ₂ Cl ₂ RT

R ² HH

R - N

J- · 1 2

wherein R, R and R have the meanings given above and X the groups

Cl, Br, OAc,

, N ₃ , NH ₂ ,

11

-0-CH, OH, -SCN, -SeCN, -SeH or -P (OCH ₂ Ph) ₂

It

and Y represents H or the same meaning as X owns.

-CH ₂ SH

CO ₂ R.

Base, for example tertiary amines, NaH, potassium tert-butoxide or a Sodium alcoholate

R ¹ ,

COOR

- 143 509820/1161

23563S2

M / 12860

In two preferred embodiments, "X" denotes the group Br and Y the groups H or Br,

There will be.so alternative ways of produced by direct fermentation Penicillins, such as penicillins G and V, to other penicillins as well. also to members of the class of the cephalosporins created. As an example of this, in which the 'above

Y means bromine, it reacts "with potassium-5-methyl-1 ^^ - thiadiazol-z-yl-thiolate, followed by the on-conversion of R, R and R; in hydrogen (if this is not already the case) to the properly thiolated in the 3-position 7-ADCA core, which is then acylated in the customary manner with 1-tetrazolylacetic acid, with Cephazolin is created. As a further example of this, the core contains 7-ADCA (T-aminodeacetoxy-cephalosporanic acid), if

■ - ■ ■ · 2 i

Y means hydrogen * If R means hydrogen and R the meaning ■

Cl ₃ C - CH ₂ O - C τ

treatment with zinc and acetic acid leads to one Ester of 7-ADCA which can be cleaved in the usual way to obtain 7-ADCA as such *

As another example, cephalexin as such is identified by the same Obtained reaction sequence, where X is bromine

1 2

Y has hydrogen and R and R, together with the nitrogen atom the group

' ^: N- ^; ■ "■■ ■■■■ ·, ■■■ -

ON-N C

CH ₃

- 144 -

50 982 0/1161

M / 12869

"mean"

AHi

The anhydropenicillin used as the original starting material in this case is obtained either by treatment from N-nitrosohetacillin (manufactured according to Belgian patent specification 765 596, Farmdoc 67,311S) to the usual Way to convert a penicillin into an anhydropenicillin, or by coupling D - (-) - 2-phenylglyc-yne (like with a carbodiimide) with the compound according to US-PS 638 of structure

followed by the formation of its ace-clay adduct and subsequent nitrosation according to the Belgian patent
765 596, Farmdoc 67 511S .. »

Suitable compounds according to the invention are used
to compounds known from the prior art in the; Prepare series of penicillins, including the penicillins themselves, as illustrated by 'the following reactionsi "". ·· -

- 145 -

50 9820/1161

M / 12860

R HH

■ N,

'R-

RHHR ¹ - N_

RHH

·

R-N-

J ^

Cl

1. NaSH Λ (crown ether) / (Cryptates) I.

or \

2. Tetramethylguanidinium- J hydrosulfide J

where R ^ is the group .C = C

'COOR ⁴ ₃

in which R is hydrogen, lower alkyl, trichloroethyl, benzhydryl or benzyl, represents \ /

'R-

S §

·

R-N-

Conversion into a penicillin according to

Wolfe et al., J. Amer. Chem. Soc. ^ 1_, 7205 (1969),

if -

COOH

"CH-,

R ² = H and R ¹ = acyl .

if R
and R ¹

χ Η
Acyl,

ie R ⁵ -COOH

according to R.D.G. Cooper and

F.L.Jose, J.Am.Chem. Soc, 92, \ 2575 (Τ570)

COOH

-N.

- 146 -

50 9820/1161

M / 12860

1HS

Crypts were discovered by B. Dietrich, J.M. Lehn and J.R. Saurage, J. Chem. Soc. (D), 1055 (1970) and crown ether by CJ. pedersen, J. Am. Chem. Soc., 89, 7017 (1967) and 92, 386, 391 (1970). Both cryptates and crown ethers are means that make it possible to use inorganic salts to be dissolved in organic solvents by complexation.

Another process leads to secopenicillins (secopenicillins) as follows:

1
R-

R ¹
t

2 H H

-N,

R ⁶ S ©

R -

R t

2 HH

0 '

egg

¹ R-

R ⁶ S

, 2 H H

_R 1. N

, SR "

N,

R ¹ - N

H H

.N,

When R is benzyl, it is made by catalytic hydrogenation converted into hydrogen, a further route to the above sulfhydryl compounds is created.

■ 5 In the above equations, -R preferably means

- 147 - '

5 0 9 8 2 0/1161

M / 12860

.-; - C = C ₄

■ = ■■ -. I-

COOR

and R, R, R ^ and. R! Have "the meanings previously used in the present application.

When treating bacterial infections in humans the new, inventive bicyclic ß-lactam antibiotics, for example the oxapenicillins and the Aza-cephalosporine in accordance with "common Method of antibiotic administration in an amount from about 5 to 200 mg / kg / day and preferably from about 5 to 20 mg / kg / day in divided doses, for example three or administered orally or parenterally four times daily. They are given in dose units, for example 125 or 250 or 500 mg of active ingredient, administered with suitable physiologically compatible carriers or binders .: The dose units are in the form of liquid \ Preparations such as solutions or suspensions or of solids in tablets or capsules “before.

The substituted azetidin-2-ones according to the invention are not effective as antibacterial agents, but they act as inhibitors of the enzyme β-lactamase and decrease so the rate of destruction of a penicillin when it is in contact is hereby needed. This is exemplified by the reduction of the minimum inhibitory concentration of ampicillin against "resistant" or ß-lactamase producing Bakr teries such as P. morganii and Ps. aeruginosa of values im Range from 500 to 1,000 γ / mT to values up to 125 γ / ml " down when using concentration of the substituted Azetidin-2-ones in the range of about 125 'γ / ml have been shown. These special values are obtained when using the connection of the structure

■■ - "■ - ■■ 148 -

-509820/1161

M / 12860

CH ₂ <|>

N O

n ⁰

COOH

0 means phenyl

- 149 -

509820/1161

Claims (1)

2 358S62 I FOLLOWED I M / 12860 Patent to ρ r ü c he 1. Method of establishing a connection of the formula - 0 ■ ^ ₃ C-HH- GH-CH C \ _CI : CHCOOH 150 ' 50 982Q / 1 181: M / 12860 ■ where R is benzyl or phenoxymethyl, _r characterized in that a compound of the formula Y / CH-CH ^ C-N- C O 2 - OR where R is benzyl _r or phenoxymethyl and R is lower alkyl, trichloroethyl, benzhydryl or benzyl, with approximately 2 2 1 mole of a compound of the formula R SLi, in which R is lower alkyl means dissolved in the compound of the formula (CH ₃ ) ₂ N - P - N (CH ₃ ) ₂ lets react. 2 *. Method according to claim 1, characterized in that that R denotes n-propyl or tert-butyl. 3. The method according to claim 2, characterized in that R ^{1 is} benzyl. 4. The method according to claim 3, records that R means methyl . - 151 '■ - characterized by 109820/1161 Μ / 12860 5. The method according to claim 3, characterized in that ' that R is tert-butyl. 5.- The method according to claim 3, characterized records that R is trichloroethyl. 7. The method according to claim 2, characterized in that that R is phenoxymethyl. 8. The method according to claim ""; 7, characterized in that that R is methyl. 9 · · Method according to claim 7, characterized in that that R. is tert-butyl. 10. The method according to claim. 7 · , thus identified records that R is trichloroethyl. · 11 .. compound of formula R ¹ - C - NH - CH CH C ι ι C - N - CHCOOH 1
wherein R is benzyl or phenoxymethyl. 12. A compound according to claim 13, wherein R is benzyl means. "'■ 13. Compound according to claim 11 /, wherein R is phenoxymethyl means. . · * ■. . . . "■ · ■""''. * ^R '" -152 · S- ■■ "" · " '■' ". 5 0 9820/116 T ' M / 12860 fSI 14. Process for the preparation of the compound of formula H ₂ N - CH CH • CHCOOH characterized in that a compound of the formula R ¹ - C - NH - CH - CH CHCOOH where R is benzyl or phenoxymethyl, with a deacylating agent implements. 15. · The method according to claim 14, characterized in that the ■ deacylating agent is S. lavendulae. 16. ■ _ The method according to claim 14, characterized in that that the deacylating agent is E. coli. 17. The method according to claim 14, characterized in that the De acyl ier longitudinal process by successive Addition of trimethylchlorosilane, phosphorus pentachloride and methanol is carried out. - - · 153 ' 509820/1161 M / 12860 18. The method according to claim.-17, characterized in that that R is benzyl. 19. The method according to claim .17, characterized; that R is phenoxymethyl. 20.- Connection of. formula \ j H ₂ N-CH-CH N - ■ CHCOOH 21. A cationic salt of the compound according to claim 20th 22. Process for the preparation of a compound of Formula; · O -NH- CH— CH C ^ ³ Il I. I ν I ^GH 3 R. - C Ν - - CHCOOH 0 \ if ■ . wherein R - C - is acyl, characterized in that one the compound of formula H ₂ N - CH-CH _G. N CHCOOH 509S20 / 1TBT M / 12860 with a carboxylic acid of the formula R - COOH or its equivalent acylated as an acylating agent for a primary amino group. 23. The method according to claim 22, characterized in that that the acylating agent is the corresponding carboxylic acid chloride, acid bromide and anhydride, the mixed one Acid anhydride with a lower alkyl ester of carbonic acid or a free acid in combination with a carbodiimide is. χ 24. Method according to claim 23, characterized records that the acyl group _Λ s R - C - the groups of the formulas Ar - Il CHC Ar-X-C-C Ii • c Ar - _wm X
Ar-NH-Cj - OH ² O I Il C-C or R ¹¹ - Il NH -.C -. or C - - 155 «- 509820/1161 M / 12860 Sf ■ -ι?
wherein R is 2,2,2-trichlpräthyl or benzyl; R hydrogen, amino, carbobenzoxyamino, phenyl, fluorine, Chlorine, bromine, iodine, hydroxy, lower alkanoyloxy or lower alkoxy; .. '. " X oxygen or sulfur; ■ R ^ and R, which can be identical or different, hydrogen, phenyl, benzyl, phenethyl or lower alkyl; R ⁷ lower alkyl; 8 9 R and R-, which can be the same or different, lower alkyl, lower alkylthio, benzylthio, cyclohexyl, cyclopentyl, Cycloheptyl, benzyl, phenethyl, phenylpropyl, furyl, thienyl, Naphthyl or Ar; R lower alkylamino, di (lower) alkylamino, cycloalkylamino having J to 7 carbon atoms including, allylamino, diallylamino, phenyl (lower) alkylamino, morpholino ^ lower (alkyl) morpholino, di (lower) alkylmorpholino, morpholino - (lower) -alkylamino, pyrrolidino, lower alkylpyrrolidino, di- (lower) -alkylpyrrolidino, N, N-hexamethyleneimino, piperidino, lower alkylpiperidino, di- (lower) -alkylpiperidino, 1,2,5,6-tetrahydropyridino, N- ( Lower) -alkylrpiperazino, N-phenylpiperazino, N- (lower) -allcyl- (lower) -alkylpiperazino, N- (lower) -alkyl-di- (lower) -alkylpiperazino _s furfurylamino, tetrahydrofurfurylamino, N- (lower) -alkyl -N-furfuryl amino, N-alkyl-N-anilino or lower alkoxyianilino; . · · '· Z, Z and Z ^ _f, which can be the same or different, lower alkyl or Ar; '·' 11 ■ ' R lower alkyl, lower cycloalkyl, naphthyl, benzyl, phenethyl or. '' - · .'-.- ·. "..-'-. 5 0.9 8 2 0/11 61 M / 12860. mean, and Ar is a monovalent residue of the formulas V W-. . R. or ■ R R ' means, 12 3
where R, R and R, which can be the same or different, mean Y / ace, chlorine, bromine, iodine, trifluoromethyl, phenyl, lower alkyl or lower alkoxy, provided that only one of the groups R ', R and R is phenyl " can mean, represent. 25., Method according to claim ~ 23, characterized records that the acyl group 1 «R-C- the formula represents wherein R is lower alkyl. 26- The method according to claim 23, characterized in s records that the acyl group 0 A II R-C- the formula. - 157 ¹ 509820/1161 Ii -c - · M / 1286Ö fSB represents in which R ^{1 is} lower alkyl and R ² and R ⁵ , which can be identical or different, represent a group selected from hydrogen and chlorine. ^: 27. The method according to claim 23 -, characterized in that the acyl group R formula o ■■■ ". ■," .. ■ Ii Il C- the Il 1 2 represents wherein R is selected lower alkyl and R is selected a group under hydrogen and chlorine mean » 28. The method according to claim '23, characterized in that the acyl group R the formula 0 Il C-NHR Il represents wherein R is lower alkyl. 29. The method according to claim 23, characterized in that the acyl group R formula - C - the 509820/1161 M / 12860 represents wherein R is lower alkyl. 30th Process according to claim 23, characterized in that the acyl group O
R - C - the formula represents wherein R is lower alkyl. 31. Process according to claim 23, characterized in that the acyl group 1 i?
RC- the formula represents. - 159 - 509820/1161 Hi 32. The method according to claim 23, characterized in that the acyl group · ' o ■ ·: ■■ -1 Il R - C- the formula - represents. · 33. The method according to claim 23, characterized in that the acyl group 1 I? R - C - the formula Il CH - σ CH, I. O - ο - II 0 represents. ' 34. The method according to claim 23, characterized in that the acyl group 0 -I Il .R - C - the formula -0 - CH ₂ -C- - 160V'- 509820/1161 ΜΛ2860 represents. Method according to claim 23, characterized in that records that the acyl group
O 1 "
RC- the formula represents. 36. The method according to claim 23, characterized in that the acyl group 1 β R - C - the formula represents wherein B represents an easily removable blocking group. 37. The method according to claim 23, characterized in that the acyl group τ 9. R-C- the formula .. "- 161». - Λ 509820/1 161 M / 12860. CHC-
« NHB represents where B. is an easily removable blocking group, selected from hydrochloric acid, carbobenzyloxy, aliphatic ß-diketones and aliphatic ß-diketoesters means. . / 38th. ' Method according to claim 23, characterized in that that the blocking group is hydrogen chloride. 39 "" method according to claim 23, characterized in that that the blocking group is methyl acetic acid ester. ". ' 40. Compound of the formula R-C-NH-CH-GH C II : ■ v / β N -CHCOOH • -: ■ -. ■ Ό.:. ■■■■■ 1 " wherein R - C - is acyl or a non-toxic, pharmaceutical tolerable salt from it. 41. A compound according to claim 40, wherein R is hydrogen and R is the groups of the formulas - 162 ¹ - 509820/1161 M / 12860 Ar - CHC -; Ar-X- O Ar O ! Il Il C - Ο-; - .. C A ⁶ _mm ' t N, Il - ϊ O π - R 10 Il .C - Il Ar. - NH - C -; - OH No. ¹ O I. Il C - C. or Ii -NH-C- or 12 " - 163 «- ·· 509820/1161 12th
wherein ■ R 2,2,2-trichloroethyl or benzyl; R is hydrogen, amino, carbobenzoxyamino; Phenyl, fluorine, Chlorine, bromine, iodine, hydroxy, lower alkanoyloxy or lower alkoxy; . · X oxygen or sulfur; ■ R ⁵ and R, which can be the same or different, are hydrogen, phenyl, benzyl, phenethyl or lower alkyl; R ⁷ lower alkyl; . 8 9 R and R-, which can be the same or different, lower alkyl, lower alkylthio, benzylthio., Cyclohexyl, cyclopentyl, Cycloheptyl, benzyl, phenethyl, phenylpropyl, furyl, thienyl, Naphthyl or Ar; . . ' ·. . R lower alkylamino, di (lower) alkylamino, cycloalkylamino of 3 to 7 carbon atoms including, allylamino, diallylamino, phenyl (lower) alkylamino, morpholino ^ lower (alkyl) morpholino, di (lower) alkylmorpholino, morpholino ^ (Lower) -alkylaraino, pyrrolidino, lower alkylpyrrolidino, di- (lower) -alkylpyrrolidino, N, N-hexamethyleneimino, piperidino, lower alkylpiperidino, di- (lower) -alkylpiporidino, 1,2,5,6-tetrahydropyridino, N- (Lower) -alkylpiperazino, N-phenylpiperazino, N- (lower) -alkyl- (lower) -alkylpiperazino, N- (lower) -alkyi-di- (lower) -alkylpiperazino, furfurylamino, tetrahydrofurfurylamino, N- (lower) - alkyl-N-furfurylamino, N-alkyl-N-anilino or lower alkoxyanilino; '' / Z, Z and Z, which can be the same or different, lower alkyl or Ar; \ ■ " II · ■ "■ · '" R lower alkyl, lower cycloalkyl, naphthyl, benzyl, Phenethyl or 5.0 Ar • - ο ·.
• ti
- C - ^ 9 8 164 V- .. 2 OViK M / 12860 ns mean, and Ar is a monovalent residue of the formulas ■ R ' means, 12 3 wherein R, R and R, the same or different no can, hydrogen, chlorine, bromine, iodine, trifluonaothyl, Phenyl, lower alkyl or lower alkoxy bodouton, vorus 12 3 suppose that only one of the groups R, "R and R Phonyl can mean represent. 42.
group R - C- the formula A compound according to claim 40, wherein the acyl represents wherein R is lower alkyl. 43.
group 1 ί R - C - the formula A compound according to claim 40, wherein the acyl -Λ.65 ¹ - 509S2Q / 1161 M / 1'2860 Il R " represents wherein R is lower alkyl and R and. R that same or can be different :, a group selected from Mean hydrogen and chlorine. 44. Compound according to claim '4G ^ wherein the acyl group' - ο- · '- ■■ - "■' -.- ■■■ 1 "
R -. C- the formula I! ~ c ■ ■ // 1 2 represents wherein R is selected lower alkyl and R is selected a group mean under hydrogen and chlorine. 45. A compound according to claim 40, wherein the ac'yl group , - ■ ■ o ■ - '■: 0 Il 1 " ^ C -NHR R-C- the formula 0 - 166 «- 509 820/1161 H / 12860 represents wherein R is lower alkyl. 46. Compound according to claim 40, wherein the acyl group
O Λ Il R-C- the formula / Λ-cH-L O - C - R Represents 0, wherein R is lower alkyl. 47. Compound according to claim '40, wherein the acyl group . OR R-C- the formula where R is lower alkyl. 48. A compound according to claim 40, wherein the acyl group .R-C- the formula - 167 '- 509820/1161 HS represents. 49 compound according to claim 40, wherein the acyl group O · 1 "R -C- the formula Cl OCH ₃ • ei - <' ^y y - CH - c represents. 50. A compound of Claim 40 wherein the acyl group 1 i? R -C-- the formula Il CH-C- O - C - CH, "Il -> . ■■ -. ο ■ represents. 51. A compound according to claim 40, wherein the acyl group _Λ 8 ■■ ..:. ■ R - C - the formula. . - 168 * - '"V- S0 98 2 0/1181 no NH ₂ represents. 52. · The D-isomer © of the prebond according to claim - 169 - € 01820/1161