CH646961A5 - DERIVATIVES OR ISOXAZOLIN-5-ACETIC ACID, METHOD FOR THE PRODUCTION THEREOF AND MICROBICIDAL COMPOSITIONS CONTAINING THE SAME. - Google Patents

Description

The present invention relates to new derivatives of iso-xazoline-5-acetic acid. Furthermore, the present invention relates to a new process for the preparation of these derivatives of isoxazoline-5-acetic acid, in which a cyclopentene substituted with a protected amino group and a protected hydroxylamine group is reacted with a ruthenium compound and an oxidizing agent, a corresponding, am Nitrogen atom of the isoxazoline ring protected derivative of isoxazoline-5-acetic acid. Through further reactions, the (aS, 5S) -a-amino-3-chloro-2-isoxazoline-5-acetic acid, which is subsequently abbreviated as (AT-125), and new derivatives and analogues of these compounds can be prepared.

The (aS, 5S) -a-amino-3-chloro-2-isoxazoline-5-acetic acid and microbiological processes for its preparation are described in U.S. Patent Nos. 3,856,807 and 3,878,047. These patents also describe the effectiveness of AT-125 against tumors and its antimicrobial activity.

The present invention relates to new derivatives of iso-xazoline-5-acetic acid, which have an activity which inhibits microbial growth.

The present invention relates to new derivatives of isoxazoline-5-acetic acid, which are characterized in that they are optical isomers of a compound of the formula I or of the formula VIII

-COR6, -C-R7 wherein

R & represents an alkyl group with 1 -8 carbon atoms, a halo-25 alkyl group with 1-3 halogen atoms and 1-5 carbon atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7-20 carbon atoms and

R7 represents an alkyl group with 1-12 carbon atoms, an aryl-30 group with 6-20 carbon atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7-20 carbon atoms, or R14 and R15 together with the nitrogen atom to which they are attached are bound, a grouping of the formula

35

40 -

45

Rs has the following meanings:

(a) a grouping of the formula R9 R11

I I

-C-C-

I I

Rio R12 in which

R9, Rio, Ri i and R12 independently of one another have the meaning of hydrogen atoms or alkyl groups with 1-5 carbon atoms or 60 (b) a grouping of the formula above

55

ch ch,

'V

65

lH2

/ CH2

nch £

9

646 961

or

(c) orthointerphenylene or

(d) represents a substituted orthointerphenylene.

Furthermore, in formula I X is a bromine atom, a chlorine atom or a grouping of the formulas

-ORi, -SRi, or-NR'R "

being in these groups

Ri represents an alkyl group with 1-12 carbon atoms, an aryl group with 6-20 carbon atoms or an aralkyl group with 7-20 carbon atoms,

R 'and R "are identical to or different from one another and denote hydrogen atoms or alkyl groups with 1-8 carbon atoms and the prerequisite is that when all the radicals R, Ri4 and Ris denote hydrogen atoms, X must not be a chlorine atom and the further requirement that when X is a chlorine atom and R is either a hydrogen atom or a lower alkyl group with 1-8 carbon atoms, neither Ri4 nor Ris may be an alkoxycarbonyl group, nor may Ri4 and Ris together with the nitrogen atom to which they are attached be one Grouping the formula

0

may form, in which Rs has the meaning given above.

In the compounds of the formula VIII, R "" is an alkyl group with 1-12 carbon atoms, a halogenated alkyl group with 1-5 carbon atoms and 1-3 halogen atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7-20 carbon atoms,

Ri4 'and Ris' represent hydrogen atoms or groups of the formula

O O

II II

-COR6, -C-Ri '

with the proviso that at least one of the substituents Ri4 'and Ris' is hydrogen, and in these groups Ró has the same meaning as in formula I and R7' an alkyl group with 1-8 carbon atoms, an aryl group with 6-20 Carbon atoms, an aralkyl group having 7-20 carbon atoms or a substituted aralkyl group having 7-20 carbon atoms, or R14 'and Ru' together with the nitrogen atom to which they are attached form a grouping of the formula wherein

Rs has the same meaning as in formula I and R13 is a hydrogen atom, an alkoxycarbonyl group,

a halogenated alkoxycarbonyl group or an aralko-5 xycarbonyl group, provided that when R13 ', Ru' and Ris are all hydrogen, R "" is not an alkyl group of 1-8 carbon atoms, and when R13 is a halogenated alkoxycarbonyl group or is an aralkoxycarbonyl group, neither the group of the io formula can be an aralkoxycarbonyl group or a halogenated aralkoxycarbonyl group, nor the group of the formula

20th

may be an aralkylcarbonyl group or a halogenated alkylcarbonyl group.

As already mentioned, the (aS, 5S) -a-amino-3-chloro-2-isoxazoline-5-acetic acid and some derivatives of this acid have already been described in the literature. The above disclaimer excludes these already known compounds from the scope of protection of the formulas I and VIII.

Those compounds of the formula I according to the invention in which at least one of the radicals Ru or Ris is a radical protecting the amino nitrogen atom and furthermore the radical R has a different meaning than that of a hydrogen atom can be prepared by using a corresponding ester of tricholomic acid Subjects to chlorination. A preferred protecting group for the amino substituent is that in which the two residues Ru and R15 together represent a phthalide group.

The present invention therefore furthermore relates to a process for the preparation of optically active isomers of the formula I '

I '

or racemic mixtures of the optically active isomers of the compounds of the formula I ', in which R' "is an alkyl group with 1-8 carbon atoms, a halo-65 alkyl group with 1-3 halogen atoms, and with 1-5 carbon atoms or an aralkyl group with 7- 20. This process is characterized in that a compound of the formula II

O

II

15 -COR6

25th

o II

-C-R7 '

35

40

60

646 961

10th

0

H NT

C02R

Mt.

ïV

ff ff

II

wherein R '"has the same meaning as in formula I' subject to chlorination.

Another object of the present invention is a process for the preparation of optically active isomers of the formula I 'and the formula VH'd

Cl

COaH

<§> -

i | H

CHaOCNQ

0

I 1 13

10th

I lc with the compound of formula Vll'd

20th

25th

0

II

■ CHaOC

V

VI ld

COaH

obtained, the compound of formula VII 'd 30 is isolated from this mixture and the benzyloxycarbonyl protective group is split off from the remaining compound of formula Ile and thus a compound of formula IIb

35

VI Jd or racemic mixtures of the optically active isomers of the compounds of the formula I 'or VH'd, where in formula I'

R '"is an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms, and with 1-5 carbon atoms or an aralkyl group with 7-20 carbon atoms.

This method is characterized in that a compound of the formula purple

40

45

Mb receives this to the ester of formula IIa

50

H

55

60

I la with a ruthenium compound in the presence of an oxidizing agent and thereby a mixture of the compound of formula Ile in which R '"has the same meaning as esterified in formula I', 65 and this ester is chlorinated to form the compound of formula I. 'submits.

From the chloroesters of the formula I 'prepared by these processes, optical isomers of the formula I "

11

646 961

sr h ntya or racemic mixtures of the optically active isomers of the compounds of the formula I "or salts of these compounds, where in formula I"

Ri represents an alkyl group with 1-12 carbon atoms, an aryl group with 6-20 carbon atoms or an aralkyl group with 7-20 carbon atoms, by the chloroesters of the formula I 'to form the corresponding free acid of the formula

Cl saponified, and this with an alkali metal salt of a mercapane of the formula

Ri-S-H

wherein Ri has the same meaning as in formula I ", to form the compound of formula I".

From the chloroesters of the formula I 'it is also possible to obtain the optically active isomers of the acids of the formula V "

no • r "

'nh,

or racemic mixtures of the optically active isomers of the acids of the formula I '"or salts of these compounds, in which formula I'"

R 'and R "are identical to or different from one another and denote hydrogen atoms or alkyl groups with 1-8 carbon atoms. In this production process, the chloroester of the formula I' is first formed to give the corresponding free acid of the formula

Cl saponified and this with ammonia or an amine

^ R '

Formula H-N <^

\ r »

wherein R 'and R "have the same meaning as in formula I'", converted to the compounds of formula V ".

The optically active isomer of an acid of the formula I can also be obtained from the esters of the formula II

Prepare 0-ch or racemic mixtures of the optically active isomer of the formula I "" or salts of this acid by using a compound of the formula II

0

wherein

R '"represents an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms and 1-5 carbon atoms or an aralkyl group with 7-20 carbon atoms, with diazomethane in a solvent with a catalytic amount of boron trifluoride di-etherate and the Amino protecting group and the group R '"split off. The acid obtained is isolated in free form or in the form of a salt.

In connection with the definition of the substituents in the derivatives according to the invention of isoxazoline-5-acetic acid of the formula I or the formula VIII, the term alkyl groups having 1-8 carbon atoms or else the term lower alkyl groups, for example methyl or ethyl - To understand propyl, butyl, pentyl, hexal, heptyl or octyl group and isomeric forms of these groups.

Examples of halogenated alkyl groups with 1-5 carbon atoms which are substituted with 1-3 halogen atoms are such methyl, ethyl, propyl, butyl and pentyl groups and their isomeric forms, with 1 to and 3 bromine -, Fluorine, chlorine or iodine atoms are substituted.

The term “aryl group” means phenyl and phenyl-containing groups which contain halogen atoms, alkoxy groups, alkyl groups and / or nitro groups.

The term "aralkyl groups" refers to benzyl, phen-ethyl, phenylpropyl, phenylbutyl, phenylpentyl, diphenylmethyl, 3,3-diphenyloctyl groups and isomeric forms thereof.

The term “substituted aralkyl groups” denotes aralkyl groups in which the phenyl ring contains substituents such as haloalkoxy groups, alkyl groups and nitro groups. Examples include p-methoxybenzyl, m-methoxybenzyl and p-nitrobenzyl.

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

646 961

12

The term “substituted orthointerphenylenes” denotes lower alkyl, lower alkoxy, halogen, nitro and cyanide substituted orthointerphenylenes. Combinations of substituents may exist, e.g. 4-propyl-2-methyl, 2-chloro-4-methyl, 3,4-diethoxy, 3-cyano-4-ethoxyphenoxy and the like. The substituted phenoxy groups contain a maximum of 10 carbon atoms.

Halogen means bromine, chlorine, fluorine and iodine.

The present invention opens a way to chemically prepare AT-125 and new analogs of this compound.

The method defined in claim 12 is advantageous because it enables complete chemically synthetic access to AT-125, which was previously only available via a microbiological method. The full process produces dl-trans-3-amino-4-hydroxy-cyclopentene which is converted in a series of reactions to give tricholomic acid derivatives, AT-125 and the bromine analogue thereof. These halogenated compounds are then converted into further analogs.

Tricholomic acid was first isolated by Takemoto et al., [See Zykugaku Zasshi, 84, 1183 and 1230 (1964)] and is an amino acid of the following structure-fighting agent (flies die after enjoying this compound) and that it has an odor which is perceived as pleasant by people [s. Iwasaki et al., Chem., Pharm. Bull 17 (5) 866-872 (1969)]. 5 In addition to the work of Takemoto et al. other methods of making tricholomic acid have been described. See, for example, [Iwasaki et al., Chem. Pharm. Bull 17 (5) 866-872 (1969)], [Iwasaki et al., Chem. Pharm. Bull 17 (5) 873-878 (1969)], [ Kamiya, T., Chem. Io Pharm. Bull 17 (5) 879-885 (1969)], [Kamiya, T., Chem. Pharm. Bull 17 (5) 886-889 (1969)], [Kamiya, T ., Chem. Pharm. Bull. 17 (5) 885-900 (1969)].

AT-125, or (a5,5S) -a-amino-3-chloro-a-isoxazoline-5-acetic acid is a tumor-inhibiting and antimicrobial agent of the general formula

20th

cl co2h h nhj c02h

This compound is known to be called a fly

30th

The novel process, which is used to produce tri-35 cholomic acid and its derivatives, can be explained schematically using the following formula scheme.

Level 2

2 parts ho

level 3

nc0ch2cc13

IVb

IVa

0

I t le h2n0

\ v Level 5 w sncoch2cci3

I lld

Level 6

Level 7

r— \ | ih / \ h | i j — v ii ",

o3 / -ch2ocno ncoch2cci3 (o) —ch2 ° cn nh2

l I lc l Hb

13

646 961

Level 8

S »M A

CHpOCNO N.

0

Il la-0

0

I ib x 1 level 12

h \ n jo lc

I lc h

"\

0

I la

Ib.

co2h

The starting materials which are used to prepare the compounds of the formula I according to the invention and also the compounds of the formula VIII according to the invention

U

R13 <^ NR * 4Rl5 ^ CO 2 Rw iRl3 ~ n

Vlld co2r

VII *

can be used have the following

NR \ 4 Ri 3

II

646 961

14

These compounds include racemic mixtures and also the optically active isomers.

In the compounds of the formula VIII, the substituents R13, Ri4 ', R15' and R "" have the meaning defined in claim 1. A specific example of compounds of formula II are those of the following structure

H

mean with 7 to and with 20 carbon atoms. R4 and Rs can also form a group of the formula together with the nitrogen atom, in which formula Rs is a group of the formula (a) Rs Ru

I I

-C-C-

I I

Rio Rl2

in which

R '"represents an alkyl group of 1-8 carbon atoms, a halogenated alkyl group of 1-3 halogen atoms and 1-5 carbon atoms, or an aralkyl group of 7-20 carbon atoms. When these compounds are reacted with diazomethane in a solvent with a catalytic amount of boron trifluoride etherate and the amino protecting group and the group R '"are split off, optically active isomers of the formula I" "or racemic mixtures of the optically active isomer of the formula I" "or salts of the compounds in question are obtained.

The compounds of the formulas IVb and IVa appearing in the reaction scheme are specific examples of compounds of the following formula IV

r3O

/ R4

I.

xr5

(IV)

wherein in this formula R3 is a hydrogen atom or a group of the formula

O

II

RaS-

O

20 wherein R'j, Rio, Rh and R12 represent hydrogen atoms or alkyl groups with I to and with 5 carbon atoms; a group (b) of the formula

25 \ / CH2 \

CH CH0

I I 2

.CH CH_

^ \ X

XCH "/

30 2

(c) means orthointerphenylene and (d) substituted orthointerphenylene.

(III) Compound, namely racemic mixtures and 35 optically active isomers of the compound of the formula

40

RcO

(III)

wherein in this formula Rc is an amino group, a dialkyl-45 amino group, a group of the formula

OH

OH

II I II I

R "óOCN- and R" 7CN-,

so where R "6 and R" 7 alkyl groups with 1 to and with 8 carbon atoms, halogenated alkyl groups with 1 to and with 5 carbon atoms and 1 to and with 3 halogen atoms, aralkyl groups with 7 to and with 20 carbon atoms, ss substituted aralkyl groups with 7 to and with 20 carbon atoms as well as the group

0

in this formula, Ra denotes an alkyl group having 1 to 8 carbon atoms, a halogenated alkyl group having 1 to 5 carbon atoms and 1 to 3 halogen atoms or an aryl group having 6 to 20 carbon atoms, and R4 and Rs are the same or different and hydrogen atoms or groups of the formulas

OO 60

II II

R "60C-, R" tC-

mean, in these formulas R "6 and R" 7 alkyl groups with 1 to and with 8 carbon atoms, halogenated 65 alkyl groups with 1 to and with 5 carbon atoms and with 1 to and with 3 halogen atoms, aralkyl groups with 7 to and with 20 carbon atoms, substituted aralkyl groups, in which group Rs is a grouping (a) of the formula

-N

R £

0

15

646961

Rs RH

I I

-C C-

Rii

Ri:

of the formula

0

-N

R,

and where Ro, Rio, Ri i and Ri: in this formula are hydrogen or alkyl groups having 1 to 5 carbon atoms, a group (b) of the formula

\ / ° \ ch ch,

/ CH \ / CH2

C 2

(c) means orthointerphenylene and (d) substituted orthointerphenylene and where R-t and Rs are the same or different and hydrogen atoms, groupings of the formulas

O O

II II

R "f, OC-, R" 7C-

mean or if the two radicals together with the nitrogen atom form a group, a group results

0

where the substituents R "6, R" 7 and Rs are the same as above and it is ensured that the groups R "6 and NR4R5 are never the same at the same time.

15 Intermediates can also be compounds which are racemic mixtures or optically active isomers of the compound of the formulas

20th

25th

R3O

n;

r30

vi

30 are generated, where R3 is as defined above.

The compound AT-125 and its novel derivatives can be prepared according to the procedure already given.

Step 1 is carried out in one of three ways, as indicated in the following reaction scheme.

Stage d

Level a / level b

■ ^ 0

HO n3 HO VI \ via

Vb

646 961

16

The epoxycyclopentene of the formula V used in this reaction scheme as a starting material can be prepared by the method described by Crandall et al.

in J. Org. Chem., 33: 423 (1968). In a preferred mode of reaction, referred to as step (a) in the previous reaction scheme, this starting material of formula V is reacted with a solution of ammonia in protic solvents, such as e.g. Water,

Methanol, ethanol or other alcohols or in non-protic solvents such as e.g. Diethyl ether, dimethyl-io formamide, tetrahydrofuran or dimethoxyethane, implemented. The reaction in the aprotic solvent is advantageously carried out in the presence of a catalyst, e.g.

dry basic alumina, as described for similar reactions by Posner and Rogers in J. Amer, is Chem. Soc., 99: 8214 (1978). The reaction in protic solvents is usually carried out at temperatures from -50 to + 50 ° C and in a concentration of epoxy from 0.01 M to 2M. The molar ratio of ammonia to epoxide is generally in the range 2 "from 1: 1 to 50: 1 and preferably in the range from 15: 1 to 20: 1. In non-protic solvents, the reaction is generally carried out at temperatures from -20 to 100 ° C. The concentration of the epoxide in these solvents is generally between 0.01 and 2M and the mol-2s ratio of ammonia to epoxide is generally in the range from 1: 1 to 20: 1 and preferably in a ratio of 1: 1 to 5: 1. The amine of formula IVb so prepared is generally isolated by evaporating the excess ammonia and solvent and a crystalline salt of the crude amine residue is formed.

These salts can be formed using a solution of an acid, such as e.g. Toluenesulfonic acid, hydrochloric acid, hydrobromic acid, hydroiodic acid and other acids in solvents such as e.g. Water, methanol, 35 ethanol, ether, 1,2-dimethoxyethane or p-dioxane. The salt is isolated by filtering and crystallizing directly or by evaporating the solvent and then crystallizing on a suitable solvent.

Alternatively, the crude amine of formula IVb can be purified by adsorbing an acid charged with acidic ion exchange resin (for example Dowex 50W-X2 (H +) or Amber-lite 1R-120 (H +) and then using a solvent, such as water, methanol, 45 ethanol, ether, tetrahydrofuran, 1,2-dimethoxyethane or p-dioxane, which is a volatile amine, such as

Contains ammonia, methylamine, diethylamine or triethylamine and then evaporates the volatile amine and the solvent.

Similarly, the salts of the hydroxyamine can be converted to the free amines by preparing a solution of the salt in a solvent such as e.g. Water, methanol, ethanol, tetrahydrofuran or 1,2-dimethoxyethane through a basic ion exchange resin, e.g.

Dowex 1-X8 (OH-) or Amberlite 1RA-400 (OH ") passes through and the eluate containing amine is evaporated.

The salts of the amine of formula IVb can also be converted to the free amine by mixing an aqueous solution of the salts with a slight molar excess of a base, e.g. Sodium or potassium hydroxide, and then the resulting aqueous solution with a salt such as e.g. Sodium chloride or sodium sulfate, and subject this solution to continuous solvent extraction using a solvent such as e.g. Methylene chloride or chloroform. The amine of formula IVb is then isolated by evaporating off the solvent.

A second process route for the production of the amine of formula IVb is shown in the formula scheme under stage b and it is the epoxide of formula V with a mixture of hydrochloric acid and 1,1,3,3-tetramethylguanidine in a solvent such as e.g. Methylene chloride, treated.

There are many references regarding the opening of epoxides with azide ions [see, for example, J. Org. Chem. 37, 1268 (1972); J. Amer. Chem. Soc. 93, 1813 (1971); J. Org. Chem. 32, 1452 (1967); J. Med. Chem. 15, 1975 (1972) and J. Amer. Chem. Soc. 94 7098 (1972)]. However, the high reactivity of the acyclic epoxide is unexpected. The use of 1,1,3,3-tetramethylguanidine to solubilize the acid ion in organic solvents has been described by Papa, in J. Org. Chem. 31, 1426 (1966). As far as we know, the specific opening of the epoxide to hydroxyazides with ammonia and 1,1,3,3-tetramethylguanidine in methylene chloride has not yet been described.

The molar ratios of hydrazonic acid and guanidine to epoxide should be in the ratio of 1: 1 to 5: 1 and from 1: 100 to 1: 1. The reaction can be carried out at temperatures between about -50 ° C and 100 ° C and in concentrations of 0.01 to 2M. Under these conditions, the dl-3,4-epoxycyclopentene becomes extremely reactive. Solvents other than methylene chloride can also be used, for example tetrahydrofuran, dimethylformamide, p-dioxane and glyme. The azide of the formula VI can be removed from the reaction mixture by customary separation techniques, for example by filtration, extraction, chromatography and combinations thereof.

An alternative procedure for step b can be shown schematically as follows:

NaNî

acid

■>

V

V! a

50

55

«0

65

In this reaction, dl-3,4-epoxycyclopentene is mixed with a mixture of sodium azide and a weak acid, e.g. Boric acid, reacted and kept in suspension in a solvent, e.g. Dimethylformamide, dimethyl sulfoxide or acetonitrile. About 1 to 5 moles of sodium azide per mole of epoxy and 1 to 5 moles of boric acid per mole of epoxy are used. The reaction mixture can be carried out between temperatures of about 0 and 125 ° C for a period of between about 1 hour and 10 days. The desired product, namely DL-trans-3-azido-4-hydroxycyclopentene of formula VI, is isolated from the reaction mixture by conventional procedures such as e.g. Extraction and chromatography and combinations thereof.

If left to stand for a long time at room temperature and if it heats up more quickly, the azides of the formulas VI and Via are partially converted into other azides. The two isomers of formula VI and Via can be separated chromatographically.

In stage c, the reduction of dl-trans-3-azido-4-hydroxycyclopentene of the formula VI to the dl-trans-3-amino-4-hydroxycyclopentene of the formula IVb takes place. This type of reaction is known in the prior art. However,

17th

646 961

gene results are more likely to be expected by using polar solvents which dissolve the strongly polar dl-trans-3-amino-4-hydroxycyclopentene. Suitable solvents are, for example, tetrahydrofuran, p-dioxane and 1,2-dimethoxyethane. The reaction can be carried out at temperatures between about -78 ° and + 50 ° C and the reaction can be between 0.01 to 2M. The amines of the formula IVb can be isolated from the reaction mixture by customary procedures, as have also been described above for step a.

The third way to connect the formula IVb,

namely stage d, the epoxide of formula I is treated with phthalimide in the presence of a base such as e.g. Potassium phthalimide or a tertiary amine in a solvent such as e.g. Dimethylformamide, dimethyl sulfoxide, tetrahydrofuran or acetonitrile implemented. The molar ratios of phthalimide and base to epoxy can be 1: 1 to 10: 1 and 1:10 to 1: 1, respectively. The reaction can be carried out at temperatures between about 0 and 125 ° C for about 1 hour to 20 days, the higher temperatures corresponding to shorter reaction times. The hydroxyphtha-limide of the formula IVb can be isolated from the reaction mixture by customary separation techniques, for example by extraction, chromatography and combinations thereof.

In step e, the phthalimide of the formula IVd is converted into the hydroxyamine of the formula IVb by treating with hydrazine or hydroxylamine in a known manner. Suitable solvents for this reaction include alcohols such as e.g. Methanol and ethanol, tetrahydrofuran, dioxane, dimethylformamide and acetonitrile. The molar ratio of phthalimide of formula IVb to hydrazine or hydroxylamine is generally 1: 1 to 1:10 and the reaction can be carried out at a temperature from -20 ° C to 100 ° C and preferably at or near room temperature. The amino alcohol of formula IVb can be isolated in the same manner as described in step a above.

In stage 2, the dl-trans-3-amino-4-hydroxycyclopentene of the formula IVb is reacted with trichloroethylchloroformate. In this step the amino group is blocked with the trichloroethoxycarbonyl group. The reaction is carried out according to procedures well known in the art and used to convert amines to urethanes. The reaction is carried out by mixing a molar ratio of trichloroethylchloramic acid ester to the compound of formula IVb in a ratio of about 1: 1 to 10: 1 at a temperature of about -20 ° C to 50 ° C for a period of 20 minutes admits up to 24 hours. The reaction is usually carried out in a solvent such as e.g. Water, methanol, methylene chloride, tetrahydrofuran or p-dioxane, which contains a dissolved or suspended base, e.g. Sodium or potassium carbonate or a tertiary amine to trap the acid released. The reaction can also be carried out in a solvent which itself serves as a base, e.g. Pyridine. The preferred solvent is water, which contains sodium carbonate. The urethane of formula IVa can be isolated from the reaction mixture using conventional procedures such as e.g. Crystallization, filtration, distillation, extraction, chromatography and combinations of these techniques.

Other halogen substituted alkyl halogen formates can be used to protect the amino group, for example iodoethyl chloroformate. The amino group can also be protected by means of an aralkoxycarbonyl group or an alkyloxycarbonyl group, for example by means of benzyloxycarbonyl and t-butyloxycarbonyl groups. Methods of using such groups to protect amines are well known in the art. For example, background information regarding the production and removal of phthalimide, p-nitrobenzyl esters of carbobenzyloxy and carbo-tert-butyloxy derivatives of amino acids from R.A. Boissonas, in the chapter "Selectively Removable Amino Protective Group used in the Synthesis of Peptide" in io "Advances in Organic Chemistry", 3: 159-190 (1963)

described. Information on the use of t-butyl-oxycarbonyl groups for protecting the amine function is also contained in the technical information document from “Aldrich” called BOC-ON (September 1976). 15 Information on the use of trichloroethoxycar bonyl groups for protecting amine functions is provided by Windholz et al. in "Tetrahedron Letters" 2555 (1967). To our knowledge, the procedure used according to the invention has not yet been used 20.

The urethanes of formula IVa can also be prepared from salts of dl-trans-3-amino-4-hydroxycyclopentene, which are obtained by treating the amine with protonic acids. The reaction is carried out by dissolving the salt in water for 2 seconds and then treating the solution with sodium carbonate and trichloroformate or another protective group as described in step 2 above. The urethanes of the formula IVa can in turn be isolated from the reaction mixture using 30 customary procedures.

Step 3 is carried out by reacting the urethane of the formula IVa with a slight molar excess of N-hydroxyphthalimide, diethyl azodicarboxylate and tri-phenyîphosphine in a solvent. The reaction is carried out at a temperature of about -50 to + 50 ° C. Suitable solvents are, for example, tetrahydrofuran, ether, 1,2-dimethoxyethane and p-dioxane. The preferred solvent is tetrahydrofuran. It should be noted that an inversion occurs in stage 3 and that the urethane of the formula II obtained in this way is a cis isomer.

With slight modifications, using other compounds obtained from the amine and other halogen-substituted alkyl haloformates, the method according to the invention can be used and these compounds instead of dl-trans-3-amino-4-hydroxycy-clopentene-N-trichloroethyl- Use urethane in level 3.

Likewise, compounds in which the amino group is protected with an aralkoxycarbonyl group can be used in stage 3. Namely for example the connections

55 0

IVe

646961

0

In this case, the implementation and also the isolation process are carried out essentially in the same way as described in connection with the above step 3.

In stage 4, the phthalimide compound of the formula Ille is converted into the hydroxylamine of the formula Illd by reacting with a slight polar excess of hydrazine hydrate. This reaction is carried out in the presence of a solvent at a temperature between -20 and + 100 ° C for a period between 1 hour and 2 days. Solvents that can be used include tetrahydrofuran, ethanol, methanol, water, p-dioxane and dimethylformamide.

The hydroxylamine of the formula IIld can be isolated from the reaction mixture by customary procedures, such as, for example, extraction, crystallization, chromatography and combinations of these methods.

In step 5 the hydroxylamine group of formula Illd is protected by reacting with an alkoxychloroformate, a haloalkoxychloroformate or an aralkoxyhalogenate such as e.g. Benzyl-oxychloroformate, reacted, whereby the car-bamate of formula IIIc is obtained.

The reaction is carried out by maintaining a molar ratio of aralkoxyhaloformate to the compound of formula IIld of about 1: 1 to 10: 1. One works in a solvent, e.g. Tetrahydrofuran, 1,2-dimethoxyethane, p-dioxane, acetonitrile or dimethylformamide, which is a tertiary base, e.g. Triethylamine contains, or in a solvent, such as pyridine, which itself acts as a base. The preferred solvent is pyridine. The reaction generally takes place at a temperature of from -20 to + 50 ° C.

Common procedures, such as Crystallization, extraction, chromatography and combinations of these methods can be used to isolate the product from the reaction mixture.

In step 6, the amino group of the compound IIIc is released from the protective group by reacting with zinc, whereby the aminocarbamate of the formula Illb is obtained. This reaction can be carried out and worked out according to various procedures.

The preferred method involves carrying out the reaction in the presence of methanesulfonic acid and methanol. The reaction is carried out at a temperature between about 0 and 50 ° C for a period of about 30 minutes to 5 hours using a molar ratio of zinc and acid to compound of formula IIIc of about 2: 1 to 50: 1.

In step 7, the aminocarbamate of the formula Illb is converted into the corresponding phthalimide of the formula purple by reacting with 2-methoxycarbonylbenzoyl chloride and triethylamine in the presence of a solvent, giving the phthalimide product of the formula purple. The reaction is carried out at a temperature between about -20 and about 50 ° C for a period of about 10 minutes to about 5 hours by using a molar ratio of 1: 1 to 10: 1 with respect to benzoyl chloride and one of 1: 1 to 20 : 1 applies to triethylamine compared to the compound of formula IIIc. Suitable solvents include tetrahydrofuran, ether, 1,2-dimethoxyethane, p-dioxane, dimethylformamide and methylene chloride. The product of the formula purple is isolated from the reaction mixture by customary procedures, for example by extraction, crystallization, chromatography or combinations of these methods. The preferred procedure is isolation by means of extraction and subsequent chromatography or crystallization.

The starting material, namely 2-methoxycarbonylbenzoyl chloride, can be prepared by a procedure described by Hoogwater et al. in "Recueil", 92, 819 (1973).

In step 8, the alkene phthalimide of the formula purple is reacted with a ruthenium compound, e.g. with ruthenium trichloride, in the presence of an oxidizing agent, e.g. Potassium or sodium iodate and a solvent can react to obtain the benzyloxycarbonyl derivative of tricholomic acid of the formula IIb. The reaction is carried out by using molar ratios of ruthenium chloride and sodium iodate to the compound of the formula purple of about 1: 1000 to 1:10 and 4: 1 to 10: 1, respectively, and a temperature in the range from 0 to 50 ° C. during a Time period from about 20 minutes to 24 hours used. Suitable solvents are, for example, water with acetone, methyl acetate, nitromethane or tert-butyl alcohol. The preferred solvent is acetone water. The product of the formula Ile can be isolated from the reaction mixture by customary procedures.

Likewise, a product of the formula is formed during the step 8 reaction.

In step 9, the compound of formula IIb is prepared by removing the protective group from the compound of formula Ile. The specific conditions for deprotection depend on the specific oxycarbonyl groups, namely on the group (Rb) which is bonded to the nitrogen atom of the tricholomic acid ring. If this group is a benzyloxy group or aralkoxycarbonyl group, deprotection can be carried out by dissolving the compound in a solvent and dissolving the solution with hydrogen in the presence of a conventional catalyst at a temperature between about 0 60 and 50 ° C at atmospheric pressure for a period of time from about 20 minutes to 2 hours. Suitable solvents are, for example, ethyl acetate, ethanol, toluene and tetrahydrofuran. Suitable catalysts are, for example, palladium, 5% palladium on carbon and palladium on barium carbonate. The product of formula IIb can be isolated by conventional procedures, e.g. by extraction, crystallization, chromatography and combinations of these procedures.

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5

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15

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646961

Alternatively, deprotection of compounds in which R13 is an alkoxycarbonyl or aryloxycarbonyl group can be carried out by operating in the presence of an acid in a solvent such as e.g. Nitromethane or methylene chloride.

When Ri3 is a haloalkoxycarbonyl group, the deprotection is preferably carried out in the presence of zinc.

In step 10, the tricholomic acid derivative of Formula IIb is converted to the ester of Formula IIa using procedures well known in the art for esterification. For example, the acid can be treated with diazoalkane or an arylated diazomethane, e.g. with diphenyldiazomethane, or by treating the compound of formula IIb with a hindered tertiary amine, e.g. N, N-diisopropyl-N-ethylamine and subsequent treatment with a benzyl halide, e.g. Diphenylmethyl chloride or p-methoxybenzyl bromide in a solvent such as e.g. Tetrahydrofuran, ethyl acetate, acetonitrile or dimethylformamide.

In stage 11, the tricholomic acid ester of formula IIa is subjected to chlorination, whereby the phthalimide isoxazole acetic acid ester of formula lc is obtained. The preferred procedure for the chlorination is to react the compound of formula II with hexamethylphosphoric triamide dichloride in the presence of a solvent. This reaction can be carried out at a temperature between about 25 and 60 ° C for a period of about 24-72 hours. The molar ratios of hexamethyl-phosphortriamide dichloride to the compound of formula IIa can be in the range from 1 to 3. Wolkoff [Can. J. Chem. Vol. 53, p. 1333 (1976)] describes a process for converting benzoyl hydrogens into their corresponding hydrazonyl halides. Attempts to convert the tricholomic acid, which contains an oxyamide group, into the corresponding hydrazonyl halide compounds

10th

however, using this procedure failed.

In step 12, the ester of formula lc is converted to the compound of formula Ib by either performing ester cleavage or removing the protecting groups from the amine. The preferred procedure is to do the ester cleavage first.

The ester cleavage in the compound of formula Ic is carried out by reacting with hydrogen halide gas in the presence of a solvent. The reaction is carried out at a temperature in the range of about 5 to about 30 ° C for a period of about 1 to about 4 hours. Suitable solvents are, for example, nitromethane, acetic acid and methylene chloride. The preferred solvent is nitromethane. The halogen portion Xi is given by the halogen hydrogen used in each case. For example, the use of hydrogen bromide, hydrogen chloride, hydrogen fluoride or hydrogen iodine produces the compounds of the formula Ib in which Xi is in each case a bromine or chlorine or fluorine or iodine atom.

The removal of the protective group from the phthalimidamic acid, which is obtained by carrying out the ester cleavage on the ester of the formula Ic, is carried out by reacting the product from the ester cleavage with hydrazine hydrate in water or alcohol. The product, namely (a5,5S) -a-amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid or AT-125 (if Xi is a chlorine atom) is isolated from the reaction mixture by conventional procedures, such as e.g. by extraction, crystallization, chromatography and combinations of these procedures.

The compounds of the formulas Ia, Ia2 and Ia3 are prepared from compounds of the formula Ia or IIa by using 35 customary procedures to replace halogen atoms with other groups. Examples of this are shown in the following reaction scheme:

20th

25th

npht iia sr 1

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20th

no '

I a, in this formula scheme Ri, R ', R "and Xi are as defined above.

Reaction of the compound of formula Ia in methanol 15 with one equivalent of sodium methoxide under an inert atmosphere at room temperature for 1 to 40 hours gives a compound of formula Ia in which Ri is a methyl group.

If a suspension of the compound of formula Ia 20 in an inert solvent, such as dimethylformamide, with an equivalent of an alkali metal salt of a mercaptan of the formula

Ri-S-M

treated, then the above-mentioned compounds of formula Iai.

If, on the other hand, a suspension of the compounds of the formula Ia in an inert solvent, such as dimethylformamide, with an equivalent of ammonia or an amine of the formula ^ -R '

H-Ni

-R "

25th

30th

treated, then you get the corresponding products of formula Ia3.

Treatment of the compounds of the formula IIa with diazomethane and a catalytic amount of boron trifluoride etherate in an ethereal solvent gives compounds of the formula Ia in which X represents a methoxy group.

The process according to the invention requires the selective removal of various protective groups in the presence of other protective groups. In general, the ease of removal of one group in the presence of another depends on the particular reagents used. With regard to the protecting groups, these can be selected by first removing the most easily removable protecting group and finally the most difficult to remove protecting group, as shown in Table 1.

Table 1

Acid (H +)

Hydrogen (H2)

Zn (metal)

Hydrazine

1. Substituted aralkoxycarbonyl (e.g. p-methoxybenzyloxycarbonyl)

Substituted aralkoxycarbonyl

Halogenated alkoxycarbonyl

-n

2.alkoxycarbonyl (e.g. t-butyloxycarbonyl)

Aralkoxycarbonyl

Substituted aralkoxycarbonyl *

3. Aralkoxycarbonyl (e.g. benzyloxycarbonyl)

4. Halogenated alkoxycarbonyl (e.g. trichloroethoxycarbonyl) *

5. -N

A

0

Alkoxycarbonyl *

Halogenated carbonyl *

0 *

-n.

Alkoxycarbonyl *

0

-n

* In general, it cannot be split off without destroying the entire molecule.

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Each step of the above process can either be carried out on racemic mixtures of the various reactants or it can be separated into the optical isomers in each step of the process according to the invention. The remaining process steps can then be carried out with optically active reactants.

The preferred method consists in the separation of the DL-trans-amino-4-hydroxy-cyclopentene of the formula IVb into its optical antipodes or its addition salts with protonic acids. The rest of the process is then carried out using the optically active isomers of the reactants.

The resolution of the racemic mixture into the optical antipodes can be carried out using suitable modifications which are well known to the person skilled in the art. For example, the compound of formula IVb can be separated into the optical antipodes by preparing salts with optically active acids, e.g. D or L tartaric acid. The L - (+) - acid gives those optical antipodes that lead to AT-125 of the natural configuration. The corresponding salts are obtained in optically pure form by repeatedly from a solvent such as e.g. Ethanol, recrystallized. A particularly effective procedure for maintaining the optically pure isomers of formula IVb, which lead to the natural AT-125, is that the racemic compound of formula IVb is dissolved in a solvent such as e.g. Methanol, first treated with deoxycholic acid and preferably crystallizing out the optical antipode which leads to the enantiomer AT-125. The mother liquors from these crystallizations are then converted to the free amine using any of the procedures described in the above reaction steps. This amine thus isolated is now substantially enriched with respect to the desired optical antipode and optical purity is achieved by the formation and recrystallization of the L-tartaric acid salt.

Since the compounds of the formula I according to the invention with R = H are amphoteric compounds, they form salts with acids, alkali metals (including ammonia), alkaline earth metals (including magnesium and aluminum) and amines. The metal salts can be prepared by dissolving the acids in water and adding a dilute metal base until the pH of the solution is 7 or 8. The metal salts include the sodium, potassium and calcium salts. Amine salts are those with organic bases, e.g. primary, secondary and tertiary mono-, di- and polyamines, which can also be made using the procedures described above or other widely used procedures. In addition, ammonium salts can be made using well known procedures. Other salts are obtained with therapeutically active bases which give the material according to the invention additional therapeutic activity. Such bases are, for example, the purine bases, e.g. Theophylline, theobromine, caffeine or derivatives of such purine bases; antihistamine bases, which are capable of forming salts with weak acids; Pyridine compounds, e.g. Nicotinic acid amide, isonicotinic acid hydrazide and the like; Phenylalkylamines, e.g. Adrenaline, ephedrine and the like; Choline and others.

Acid salts can be prepared by neutralizing the compounds of formula I with the appropriate acid to a pH below about 7.0 and preferably to a pH between 2 and 6. Suitable acids for this purpose are, for example, hydrochloric acid,

Sulfuric acid, phosphoric acid, sulfamic acid, hydrobromic acid and the like. Acid and base salts of the compounds can be used as starting materials for the same biological purposes.

The compounds of formulas I and Vlld prevent the growth of microorganisms in different environments. For example, AT-125 is effective against bacteria and Escherichia coli and can be used to reduce, stop and eliminate slime production in papermaking systems. AT-125 can also be used to extend the life of cultures of Trichomonas foetus, Trichomonas hominis and Trichomonas vaginalis by freeing them from Escherichia coli contamination. In addition, AT-125 can be used as an antifungal agent in the upper leather of shoes, as described in U.S. Patent No. 3,130,505. Since AT-125 is effective against Bacillus subtilis, it can also be used to suppress or prevent undesirable odors in fish or in fish storage containers, because these undesirable odors are generally caused by the subtilis bacillus mentioned above.

Furthermore, AT-125 can also be used to clean laboratory tables and equipment in laboratories for fungal tests.

The compounds of formula I are also effective to treat bacterial infections and tumors in mammals, including humans.

For this purpose, the compounds according to the invention are used in corresponding pharmaceutical preparations which are suitable for administration to humans and animals, such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, eye drops, solutions or suspensions for the oral administration and as water-in-oil emulsions. These preparations contain suitable amounts of the compound of formula I.

Either solid or liquid unit dosage forms are used for oral administration. For the manufacture of solid dosage forms, e.g. Tablets, the compounds of formula I with usual ingredients, such as Talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, acacia syrup, methyl cellulose and functionally similar materials mixed as pharmaceutical diluent or carrier materials. Capsules are produced by mixing the compound according to the invention with an inert pharmaceutical diluent and filler and pouring the mixture into a hard gelatin capsule of a suitable size. Soft gelatin capsules are made by mechanically encapsulating a slurry of the compound of the invention in an acceptable vegetable fat, a light liquid aselin (paraffin oil) or other inert oily substance.

Liquid unit dosage forms for oral administration, e.g. Syrups, elixirs and suspensions can be made. The water-soluble forms can be dissolved in an aqueous delivery aid along with sugar, flavoring, flavoring and preserving agents to give a syrup. An elixir is made by adding a water-alcohol (ethanol) delivery aid along with suitable sweeteners such as e.g. Sugar and saccharin, used together with flavoring agents.

Suspensions can be made with an aqueous delivery aid with the help of s

io

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35

40

45

50

55

60

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646 961

a suspending agent, e.g. Acacia syrup, tragacanth, methyl cellulose and the like.

For parenteral administration, liquid unit dosage forms are prepared using the compound of the invention and a sterile administration aid, with water being preferred. Depending on the type of administration aid and the concentration used, the compound according to the invention can be present in the administration aid either in suspended or dissolved form. In the preparation of solutions, the compound according to the invention can be dissolved in water and subjected to sterile filtration for the injection before it is filled into a suitable ampoule or a suitable vial and sealed. Aids such as e.g. Local anesthetics, preservatives and buffering agents are dissolved together with the substance according to the invention in the administration aid. To increase the stability of the compositions, they can be frozen after filling the vial and the water can be removed under vacuum. The dry lyophilized powder is then sealed in the vial and water is provided for injection in a second vial to reconstitute the solution before use. Parenteral suspensions can be made by following essentially the same procedure, except that the compound is suspended in the support instead of being dissolved and the sterilization cannot be carried out by filtration. The compound of the invention can be sterilized by exposure to ethylene oxide before suspending in the sterile delivery aid. A surface-active agent or a wetting agent can advantageously be incorporated into the composition in order to facilitate the uniform distribution of the compound according to the invention.

In addition, rectal suppositories can be used to administer the active compound according to the invention. This dosage form is of particular interest to mammals that cannot be appropriately treated by other dosage forms, e.g. by oral treatment or by blowing, i.e. in the case of small children or debilitated people. The active compound can be incorporated into any known supositional support material using procedures known in the art. Examples of such carrier material bases are, for example, cocoa butter, polyethylene glycols (carbo-waxe), polyethylene sorbitol monostearate and mixtures of these materials with other compatible materials in order to change the melting point or the rate of dissolution. Such rectal suppositories can weigh between about 1 to 2.5 g.

The term "unit dosage form" as used in this specification refers to physically discrete units that are suitable for use as unit dosages in human patients and animals, each unit containing a predetermined amount of the active material so calculated is that the desired therapeutic effect is achieved together with the required pharmaceutical dilution with carrier material or administration aids. The specifications for the novel unit dosage forms of the present invention are given and are directly dependent on the following circumstances, namely: (a) the particular characteristic of the active material and the particular effect that is to be achieved and (b) that according to the prior art There are inherent limitations in the art to formulating such active materials for use in humans and animals, as will be shown in detail in this specification, and with other objects of the invention being achieved. Examples of suitable unit dosage forms which are in accordance with the invention are tablets, capsules, pills, suppositories, powder packs, waffles, granules, trops, teaspoon fillings, tablespoon fillings, dropwise dosing, io ampoules, vials (serum vials), aerosols with measured delivery, separated multiple of the above dosage forms and other forms than described here.

An effective amount of the compound of the invention is used in the treatment. The dosage of the compound according to the invention for the treatment depends on many factors which are well known to the person skilled in the art. Such factors include, for example, the route of administration and the effectiveness of the particular compound. A dosage amount for humans from about 2 to about 200 mg of the compound according to the invention in a single dose administered parenterally or in the compositions according to the invention are effective for the treatment of tumors and bacterial infections. In particular, the single dosage ranges from about 5 to about 50 mg of the compound. The oral and rectal doses range from about 5 to about 500 mg in a single dose. In particular, the single dose ranges from about 10 to about 100 mg of the compound.

Preferred embodiments of the production of AT-125 and its analogs are described in more detail below. It will be quite clear to the person skilled in the art that variations in the production of AT-125 as well as the analogs and the precursors are entirely possible, such as e.g. 35 the change in reaction conditions and reaction techniques.

The preparation of racemic mixtures is explained below.

40 preparation I

Preparation of dl-3,4-epoxycyclopentene.

Cl

CH3CO3H V ^ -

CH2CI2 \

NazCCb \ f

V

An amount of 45 g (0.68 mol) of cyclopentadiene is dissolved in 750 ml of methylene chloride and 290 g of anhydrous sodium carbonate are added. The mixture is added dropwise with vigorous stirring over 40 minutes with 110 ml of 6.3M peracetic acid (to which 1 g of sodium acetate was added to neutralize any residual sulfuric acid) by keeping the temperature below 20 ° C by using an ice bath applied. The reaction was then stirred at 20 to 25 ° C for an additional 3 hours. The reaction was then filtered and 1 to 2 g of sodium carbonate was added to the filtrate. The filtrate was distilled by initially distilling at 35 mm pressure and finally at 20 mm pressure and the distillation temperature was ~ 30 ° C (the heating bath temperature was never above 40-45 ° C) and thus 21.8 g dl- 3,4-epoxycy-65 clopentene.

The NMR spectrum recorded in deuterochloroform gave the following signals: 8: 2.1-2.7 (m, 2H); 3.6-4.1 (m, 2H), 5.7-6.3 (m, 2H).

50

55

23

646 961

Preparation II

Production of dl-trans-azid-4-hydroxycyclopentene.

(a)

HN3 (Me2N); C = NH CH2CI2

VI

An amount of 90 g (1.38 mol) of sodium azide was added to 90 ml of water and 300 ml of methylene chloride. The mixture was stirred vigorously and cooled to less than 10 ° C and treated with 19 ml of sulfuric acid. After 30 minutes, the methylene chloride layer is decanted and dried over sodium sulfate. The solution is then treated with 8 g (0.07 mol) of 1,1,3,3-tetramethylguanidine dissolved in 30 ml of methylene chloride. The solution thus obtained is treated with the crude methylene chloride solution of 3,4-epoxycyclopentene, prepared from 0.68 mol of cyclopentadiene. After 2½ hours the solvent is evaporated in vacuo to give DL-trans-azide-4-hydroxycyclopentene.

Preparation III

Production of dl-trans-azid-4-hydroxycyclopentene and dl-trans-azid-2-hydroxycyclopentene

NaN3

The two isomers are separated by chromatography over two linearly connected silica gel columns 60 from E. Merck, size B. The columns are eluted with (40-60) ethyl acetate Skellysolve B and 30 ml-5 fractions are collected.

dl- (Trans-3-amino-4-hydroxy-cyclopentene gave the following analysis values:

NMR (CDCb, 5): 1.95-3.05 (p., ^ Hz), 3.5 (OH), 4.0-4.5 (m, 10 CHOH, CHNs), 5.6-6 , 2 (m, CH = CH).

IR (film, cm-1): 3300 (OH), 2090 (azide).

dl-Trans-3-amino-2-hydroxy-cyclopentene gave the following analysis values:

15

NMR (CDCb, 5): 1.9-2.2 (m, CH2), 3.85-4.2 (OH), 4.25-4.7 and 4.7-5.1 (m, CHOH , CHNj), 5.75-6.2 (m, CH = CH). IR (film, cm-1): 3300 (OH), 2090 (azide).

20 Thin layer chromatography on silica gel 60: the Rf value of the isomer IVc is 0.60 and the Rf value of the isomer VI is 0.39 in ethyl acetate Skellysolve B (40:60).

Preparation IV

25 Preparation of dl-trans-3-amino-4-hydroxy-cyclopentene

30th

DMF H3BO4

0H

VI

LAH THF

NH:

->

0H

IVb

Via

In an alternative procedure, the crude methylene chloride solution of DL-3,4-epoxycyclopentene, made from 36.6 ml of cyclopentadiene, is concentrated under reduced pressure of ~ 50 mm at a temperature of less than 20 ° C. The residue is added to a mixture of 8 g (129 mM) sodium azide and 8 g (129 mM) boric acid suspended in 200 ml dry dimethylformamide. The mixture was stirred at 25 ° C for 17 hours. The reaction material is then allowed to distribute between water and ethyl acetate. The ethyl acetate layer is washed with water, dried over magnesium sulfate and evaporated in vacuo at 25 ° C. The crude residue is combined with that from a previous 10 mmol batch and chromatographed on two linearly connected silica gel 60 chromatography columns from E. Merck, size B. These columns are eluted with (40-60) ethyl acetate Skellysolve B and 30 ml fractions are collected. Fractions 10-13 are combined and evaporated to give 2.64 g of dl-trans-azide-4-hydroxycyclopentene in the form of a pale yellow oil. The total yield based on cyclopentadiene is 45%.

When left standing, dl-trans-azid-4-hydroxycy-clopentene partially rearranged into dl-trans-azid-2-hydroxycyclopentene.

(a) A quantity of 2.2 g (57.9 mmol) of lithium aluminum hydride is added to 90 ml of ether and the cooled suspension is stirred in an ice bath. 3.62 g of pure DL-trans-azide-4-hydroxycyclopentene, dissolved in 5 ml of ether, are added to the above suspension 40 sion over 15 minutes and an exothermic reaction with vigorous gas evolution occurs. After stirring for 2 hours, the reaction is carefully treated with 2.2 ml of water, 2.2 ml of 15% sodium hydro-45xide solution and 2.2 ml of water and initially a violently exothermic reaction with a large gas evolution occurs. After stirring for a further 15 minutes, the solids are removed by filtration. The filtrate was evaporated in vacuo, leaving 1.96 g of dl-trans-3-50 amino-4-hydroxycyclopentene, which solidified on standing to a low-melting solid. This material is extremely water soluble. The analytical values were as follows:

55 NMR (CDCb, S): 1.8-2.9 (m, CHz), 3.2 (OH, NHz), 3.5-4.2 (m, CHOH, CHNH2), 5.4-5 , 9 (m, CH = CH).

TLC (silica gel 60): Rf = 0.24 in (1-20-80) conc. NH4OH-Me-OH-CH2CI2.

60 (b) The entire crude product of dl-trans-azide-4-hydroxy-cyclopentene from a 0.68M batch of azide preparation supported with 1,1,3,3-tetra-methylguanidine (preparation III) is dissolved in 100 ml of tetrahydrofuran and the solution is added over 30 minutes to a suspension of 25.8 g (0.68 65 mol) of lithium aluminum hydride in 1 liter of tetrahydrofuran. The temperature of the reaction mixture is kept below 5 ° C by using an ice salt bath during the addition. The reaction then turns on

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24th

Allow to warm to 25 ° C. and stir for 8 hours. The reaction mixture is then again cooled to 0 ° C. and treated successively with 25 ml (an exothermic reaction with vigorous gas evolution occurs), 25 ml of 15% sodium hydroxide solution and 25 ml of water. After stirring for half an hour, the mixture is filtered. The solids are washed with tetrahydrofuran. The combined filtrates and washing solutions are evaporated in vacuo, leaving 47.8 g of crude dl-trans-3-amino-4-hydroxycyclopentene.

Preparation V

Preparation of dl-trans-3-amino-4-hydroxycyclopentene-toluenesulfonic acid salt

(+)

-> (+)

nh;

oh ho nh3 + -os

II

0

ch:

! Vb

An amount of 22.17 g of crude dl-trans-3-amino-4-hydroxy-cyclopentene dissolved in about 200 ml of tetrahydrofuran is treated with a saturated tetrahydrofuran solution containing 40 g (0.21 mol) of p-toluenesulfonic acid monohydrate. Crystals quickly form. They are removed by filtration after a few minutes and 14.5 g of dl-trans-3-amino-4-hydroxycyclopentene-toluenesulfonic acid salt are obtained with a melting point of 180-182 ° C. The mother liquors are concentrated and the mixture is cooled, so that more are obtained

IVbi

18.9 g of dl-trans-3-amino-4-hydroxycyclopentene-toluenesulphonic acid salt in the form of crystals with a melting point of 177-181 ° C., giving a total yield of 33.4 g.

Preparation VI

Preparation of dl-3-amino-4-hydroxycyclopentene from the toluenesulfonic acid salt

© G

o II

os

II

0

ch-

Amberlite IRA-400

nh2

Oh

IVbi

A quantity of 150 ml of Amberlite resin IRA-400 in the chloride form is washed 6 times in succession in a glass sinter filter funnel with 150 ml of N sodium hydroxide solution, 3 x 150 ml of water and 3 x 150 ml of methanol. The resin beads are then slurry packed in a chromatography column containing methanol. An amount of 10.95 g (40.4 mmol) of dl-trans-3-amino-4-hydroxycyclopentene-toluenesulfonate, dissolved in a minimal amount of methanol, is then applied to the top of the column and the column is filled with 750 ml Methanol eluted. Evaporation of the entire eluate gives 4.2 g of an oil which solidifies when left to stand under vacuum overnight, whereby dl-3-amino-4-hydroxy-

->

Vb cyclopentene with a melting point of 47-50 ° C. The analytical results were as follows:

45

NMR (CDCb, 8): 1.8-2.9 (m, CH :), 3.2 (OH, NH :), 3.5-4.2 (m, CHOH, CHNH :), 5.4 -5.9 (m, CH = CH). TLC (silica gel 60): Rf - 0.24 in (1-20-80) conc. NH-tOH-MeOH-CHz-Ch. Designated with potassium permanganate-50 spray reagent.

Preparation VII

Preparation of dl-trans-3-amino-4-hydroxycyclopentene 55 of the formula IVb and the p-toluenesulfonate salts

1.NHj, MeOH

2. pTSA

OH

NH3 + -os

"0-

ch:

0

(V)

An amount of 40 ml of dry methanol is cooled in an ice bath and ammonia gas is bubbled through to make a saturated solution. 500 ml of cyclopentene epoxide are added to this ammonia solution. The reaction

25th

646 961

mixture is kept at 0 ° C for 24 hours and then at 25 ° C for 64 hours. The reaction solution is then concentrated in vacuo and the residue is treated with 5-10 ml of tetrahydrofuran. The insoluble material is removed by centrifugation to give a solution of the hydroxyamine of formula IVb.

This is then treated in a saturated solution of p-toluenesulfonic acid in tetrahydrofuran until the solution containing the amine becomes acidic. The voluminous crystalline

Precipitation is collected by filtration and dried. 680 mg of dl-trans-3-amino-4-hydroxy-cyclopentene-toluenesulfonic acid salt are obtained.

Preparation VIII

Preparation of dl-trans-3-amino-4-hydroxycyclopentene of the formula IVb and the corresponding p-toluenesulfonic acid salt.

1. NHjOH "i.- ™

a ©

^ "-0-

2. p-TSA ^ NH3 "0S - (()} - CH;

O

V! Vbj.

An amount of 1.97 g (24 mmol) of distilled expoxycyclopentene is added to an ice-cold solution of 1.97 g (39.4 mmol) of ammonium chloride in 20 ml of concentrated ammonium hydroxide. After stirring for half an hour, the reaction mixture is warmed up from 25 ° C. and stirring is continued for a further 18 hours. The reaction mixture is then extracted with ether to remove the less polar contaminants. The aqueous layer is then saturated with sodium chloride and extracted continuously with methylene chloride. The methylene chloride is dried over sodium sulfate and concentrated in vacuo, leaving 1.15 g of dl-trans-3-amino-4-hydroxycyclopentene, which are treated with 1.54 g (8.1 mmol) of toluenesulfonic acid dissolved in tetrahydrofuran, whereby 1 , 47 g of dl-trans-3-amino-4-hydroxycyclopentene toluenesulfonate is obtained.

Then, according to a similar procedure

Preparation VII, but using the appropriate protonic acid instead of toluenesulfonic acid, the racemic mixtures of:

di-p-tolyl-L-tartaric acid salt with a melting point of 185-190 ° C (decomposition),

di-benzoyl-L-tartaric acid salt with a melting point of 181-183 ° C (decomposition),

the 1-mandelic acid salt with a melting point of 116-117 ° C, and the 2-pyrrolidone-5-carboxylic acid salt with a melting point of 164-172 ° C.

Preparation IX

Production of dl-trans-4-hydroxy-3-phthalimidcyclopentene

35

40

"C0C1

OH OH

0

IVb

30 g of dl-trans-4-hydroxy-3-amino-cyclopentene are dissolved in 450 ml of tetrahydrofuran and 87 ml (0.60 mol) of triethylamine. The solution is cooled to 0 ° C. and treated dropwise with 59.6 g (0.3 mol) of o-methoxy-car-bonylbenzoyl chloride over a period of 50 minutes. The reaction is then allowed to warm to 25 ° C. After 66 hours, the reaction mixture is partitioned between ethyl acetate and water. The ethyl acetate phase is dried over magnesium sulfate and distilled off in vacuo to give 62.4 g of residual oil. This oil is spread over 3 kg of sili

[Vd cagel 60 chromatographed by eluting with acetone-methylene chloride mixtures using 61 with a mixture of 10:90, 2.5 l in a ratio of 15:85 and 11 liters in a ratio of 20:80. 200 fractions of 50 ml each are collected. The product is in fractions 38-61. The product, namely dl-trans-4-hydroxy-4-phtha limidcyclopentene, crystallized from these fractions in a yield of 25.9 g.

A process for the preparation of the O-methoxycarbonyl-benzoyl chloride is described by Hoogwater et al. in Ree. Trav. Chim. Pays-Bas, 92 (1973).

60

65

646961

26

Preparation X

Preparation of dl-trans-4-hydroxy-3-phthalimide cyclopentene

0

ö OS 0

iVbx

CH *

An amount of 31.3 g (0.115 mol) of dl-trans-3-amino-4-hydroxycyclopentene-toluenesulfonic acid salt is dissolved in 240 ml of tetrahydrofuran and 47 ml (0.46 mol) of triethylamine. The solution thus obtained is cooled to 5 ° C. with stirring and treated dropwise with 22.8 g (0.115 mol) of o-methoxycarbonylbenzoyl chloride. The reaction mixture is then allowed to warm up to 25 ° C. After 48 hours, the reaction mixture is partitioned between ethyl acetate and water. The ethyl acetate layer is dried over magnesium sulfate and concentrated in vacuo, leaving 29.6 g of a residue. This residue is chromatographed on silica gel 60

graphed by eluting with acetone-methylene chloride mixtures, with 5 1 of a mixture of 10:90 and 7.51 of a mixture of 20:80. Fractions of 300 ml each are collected. The product is isolated in the 20 fractions 25-30, which crystallize on standing, giving a yield of 14.2 g of dl-trans-4-hydroxy-3-phthalimide-cyclopentene with a melting point of 114-116 ° C.

25 preparation XI

Production of dl-trans-4-hydroxy-3-phthalimide-cyclopentene

V

2.9 g of crude dl-3,4-epoxycyclopentene is added to a solution of 7.2 g (49 mmol) of phthalimide and 1.43 g (7.7 mmol) of potassium phthalimide in 23 ml of dry dimethylformamide. After stirring at 25 ° C. for 114 hours, the reaction mixture is treated with 0.5 ml (8.3 mmol) of acetic acid and stirring is continued for one hour. The reaction mixture is then evaporated in vacuo and the residue is chromatographed on 300 g of silica gel 60 and eluted with acetone-methylene chloride in a ratio of 10:90. 50 ml fractions are collected. The product is in fractions 25-30 and it crystallizes on standing, giving 0.58 g of dl-trans-hydroxy-3-phthalimide-cyclopentene. receives.

The analytical values were as follows:

NMR (CDCb 5): 2.1-3.4 (m, CH2), 3.7 (OH), 4.6-5.0 (m, CHOH), 5.0-5.3 (m, CHN ), 5.5-6.2 (m, CH = CH), 7.7 (s, ArH).

TLC (silica gel 60): Rf = 0.32 when using ethyl acetate-Skellysolve B in the ratio 40:60 as eluent and Rf = 0.71 when using acetone-methylene chloride in the ratio of 15:85 as eluent.

The elementary analysis calculated for CBHiiNOs gave the following values:

Calculated: C 68.14; Found: C 68.19;

H 4.80 H 4.86.

Using the same procedure as in Example 6, but using the appropriate substituted 55-O-methoxycarbonylbenzoylchloride or 3-methoxycarbonylpropionylchloride instead of O-methoxycarbononylbenzoyl chloride, the corresponding compounds, namely:

dl-Trans-4-hydroxy-3 dl-Trans-4-hydroxy-3 dl-Trans-4-hydroxy-3 65 dl-Trans-4-hydroxy-3-pentene, and dl-Trans-4-hydroxy-3 -hexahydrophthalimide cyclopentene obtained.

(3-nitrophthalimide) -cyclopentene, (4-methyl-phthalimide) -cyclopentene, succinimide-cyclopentene, (2,3-dimethylsuccinimide) -cyclo-

27th

646961

Preparation XII

IVd IVb

An amount of 4.07 g (17.67 mmol) of dl-trans-4-hydroxy-3-phthalimide cyclopentene is dissolved in 65 ml of tetrahydrofuran and 65 ml of ethyl alcohol. The solution is treated with 0.94 ml (0.97 g, 19.44 mmol) of hydrazine hydrate.

A precipitate forms after 10 minutes. After 2 hours the reaction mixture is evaporated in vacuo (the pressure is less than 25 mm and the temperature is -30 ° C). The residue is treated with methylene chloride and filtered to remove insoluble phthalhydrazide.

The filtrate is concentrated in vacuo, leaving dl-trans-i »3-amino-4-hydroxy-cyclopentene in the form of a solid.

Preparation XIII

Preparation of dl-trans-3-amino-4-hydroxycyclopentene-is N-trichloroethyl urethane

O

II

CICOCH2CCI3 NazCCb

O ^ hÜ

£ ^ NCOCHsCCl:

HO

Vb

IVa

An amount of 9.9 g of crude dl-trans-3-amino-4-hydroxy-cyclopentene (prepared as in preparation IVa) is dissolved in 75 ml of water and the mixture is treated with 10.6 (100 mmol) sodium carbonate. The mixture is then cooled to less than 10 ° C in an ice bath and treated dropwise with vigorous stirring with 10.6 (50 mmol) trichloroethylchloroformate over 30 minutes. After 3 hours the reaction mixture is acidified with cold concentrated hydrochloric acid. The aqueous mixture is extracted three times with methylene chloride. The methylene chloride solutions are washed with water and dried over sodium sulfate and finally evaporated in vacuo, leaving 13.2 g of a brown oil. The oil is chromatographed over 750 g of lithium dioxide and eluted with acetone-methylene chloride in a ratio of 10:90. Fractions of 50 ml each are collected. The

(b)

Product occurs in fractions 55-85, as it turns out from thin layer chromatography. Concentration of these fractions gives 5.0 g of trans-3-amino-4-hydroxy-cyclopentene-N-trichloroethyl-urethane from crystalline-30 lineal solid with a melting point of 100-103 ° C. The analytical values were as follows:

NMR (CDCb, S): 1.9-3.1 (m.CH :), 4.0-4.65 (m, 3H), 4.74 (s, OCH2), 5.45-6.1 (m, CH = CH), 6.5-7.0 (NH). 35 TLC (silica gel 60): Rf = 0.5 when using an eluent of acetone-methylene chloride in the ratio 10:90.

Preparation XIV

40 Preparation of dl-trans-3-amino-4-hydroxycyclopentene

N-trichloroethyl urethane.

>

IVbt

0

Hii ncoch2cci3

! Va

An amount of 13.55 g (50 mmol) of trans-3-amino-4-hydroxycyclopentene-toluenesulfonic acid salt is dissolved in 75 ml of water. The solution is treated with 10.6 g (10.0 mmol) of sodium carbonate and cooled in an ice bath and then dropwise added 10.6 g (50 mmol) of trichloroethylchloroformate while stirring vigorously. After 75 minutes the reaction mixture is partitioned between water and methylene chloride. The aqueous layer is separated off and extracted again twice with methylene chloride. The combined methylene chloride phases are dried over sodium sulfate and concentrated in vacuo. They form during the narrowing

Crystals of trans-3-amino-4-hydroxy-cyclopentene-N-trichloroethyl-urethane. These crystals are collected three times, each time the crystals are washed with ether

60 see. The first crystal fraction weighed 6.12 g and showed a melting point of 105.5-106 ° C. The second crystal fraction weighed 4.35 g and showed a melting point of 105-106 ° C and the third crystal fraction weighed 1.28 g and showed a melting point of 104-105 ° C.

65 Using the same procedure as in preparation XIII, but using the appropriate halogen-substituted alkyl haloformate or corresponding activated alkyl carbonate instead of tri-

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28

chloroethyl formate, the following compounds were obtained:

dl-trans-3-amino-4-hydroxy-cyclopentene-N-t-butyl urethane,

dl-trans-3-amino-4-hydroxy-cyclopentene-N-p-methoxy-

benzyl urethane,

dl-Trans-3-amino-4-hydroxy-cyclopentene-N-diphenyl-methyl-urethane, and dl-Trans-3-amino-4-hydroxycyclopentene-N- [2-iodoethyl] -ure-than.

Using the same procedure as in preparation XIII, but using benzyloxy-chloroformate, dl-trans-3- (0-benzylcarbamoyl) -4-hydroxycyclopentene is resolved after chromatography

Obtain silica gel with 85% methylene chloride / acetone. The analytical values were as follows:

NMR (CDCb) = 2.1-2.9 (m, CH2), 3.8-4.6 (m, 3H), 5.02 (5, s OCH2), 5.3-5.0 (m , 3H), 7.22 (s, Ph).

TLC: Rf = 0.58 (15% acetone / methylene chloride).

The optically active (d) trans-5- (0-benzylcarbamoyl) -4-hydroxy-cyclopentene is also prepared in the same way, 10 using the d-starting material instead of the racemic starting material.

Preparation XV

Preparation of dI-2,2,2-trichloroethyl-cis-5- (phthalimide-is oxy) -2-cyclopentene-l-carbamate

HO

0 H | l

NCOCH2CC

! Ve

(C »Hs) JP

(= NC02C2HS) 2 THF

An amount of 11.75 g (42.8 mmol) of dl-trans-3-amino-4-hydroxy-cyclopentene-N-trichloroethyl urethane, an amount of 7.86 g (48.3 mmol) of N-hydroxyphthalimide and an amount of 12.63 g (48.3 mmol) of triphenylphosphine are dissolved in 210 ml of dry tetrahydrofuran. 9.21 g (53.1 mmol) of diethyl azodicarboxylate is added dropwise to the solution with stirring over a period of 15 minutes, while keeping the temperatures below 35 ° C. using an ice bath. The reaction mixture is then stirred at 25 ° C for one hour and then concentrated in vacuo using a pressure less than 25 mm and a temperature of about 30 ° C. The residue is treated with approximately 100 ml of a 10:60 mixture of ethyl acetate / Skellysolve B and the precipitate of triphenylphosphine oxide is removed by filtration after about 15 minutes. The residue is applied to the top of a column packed with 1 kg of silica gel 60, which is eluted with 3 liters of ethyl acetate / Skellysolve B 40:60 and then 50:50. Three hundred ml fractions are collected. Fractions 14-19 contained the pure dl-2,2,2-trichloroethyl-cis-5- (phthalimidoxy) -2-cyclopentene-l-carbamate, as it turned out from the thin layer chromatography. When concentrated, 6.24 g of kri-

45 stalls of a first crystal fraction with a melting point of 138.5-139.5 ° C, 2.8 g of a second crystal fraction with a melting point of 138.5-139.5 ° C and a residue of 2.13 g remained Evaporation to dryness. Fractions 20-24 showed, based on thin-layer chromatography 50 tography, that they contained a more polar impurity. These fractions were combined with the 2.13 g of the residue from the above fractions and chromatographed again over 750 g of silica gel 60 by extracting with ethyl acetate / benzene (10:90). 55 fractions of 300 ml each were collected. Fractions 8-13 contained the desired product based on analysis by thin layer chromatography. Concentration of these fractions gave two more crystal fractions of dl-2,2,2-trichloroethyl-cis-5- (phthalimidoxy) -2-cyclopentene-1-carbamate of 60, 2.75 g and 2.62 g and the melting points were 138.5-139.5 and 137.5-138.5 ° C. The total yield was 14.41 g.

The analytical values were as follows: NMR (CDCb, S): 2.65-3.0 (m, CH :), 4.77 (s, OCH2) 4.7-5.2 (m, 2H), 5 , 7-6.1 (m, CH = CH), 6.35-6.37 (NH), (s, ArH). 6s TCL (silica gel 60): Rf = 0.55 using ethyl acetate / Skellysolve B 40:60 and Rf = 0.47 using ethyl acetate-benzene (10:90) as eluent.

29

646 961

The elementary analysis calculated for C16H13CI3N2O5 gave the following values:

Calculated: C 45.79; Found: C 45.92;

H 3.12; H 3.13;

N 6.68; N 6.62;

Cl 25.35 Cl 25.47.

Using the same procedure as in preparation XV, but using the appropriate dl-trans-3-amino-4-hydroxycyclopentene-N-haloalkyl-urethane instead of dl-trans-3-amino-4-hydroxycyclopentene-N- trichloroethyl urethane the following compounds are obtained:

dl-butyI-cis-5- (phthalimidoxy) -2-cyclopentene-1-carbamate, dl-p-methoxybenzyl-cis-5- (phthalimidoxy) -2-cyclopentene-1-carbamate,

dl-DiphenylmethyI-cis-5- (phthalimidoxy) -2-cyclopentene-1-carbamate.

Using the same procedure as in Preparation XV, but using the appropriate N-hydroxyphthalimides and N-hydroxysuccinimides instead of N-hydroxyphthalimide, the following compounds are obtained:

dl- (2,2,2-trichloroethyI) -cis-5- (3-nitrophthalimidoxy) -2-cyclopentene-1-carbamate,

dl- (2,2,2-trichloroethyl) -cis-5- (3-nitrophthalimide) -2-cyclopentene-1-carbamate, s dl- (2,2,2-trichloroethyl) -cis-5- ( succinimidoxy) -2-cyclopentene-1-carbamate, and dl- (2,2,2-trichloroethyl) cis-5- (2,3-dimethylsuccinimidoxy) -2-cyclopentene-1-carbamate.

10 Using the same procedure as in preparation XV, but using different alkyl methanes and substituted n-hydroxyphthalimides and n-hydroxysuccinimides, the following compounds are obtained:

15

dl-benzyl-cis-5- (3 -nitrophthalimidoxy) -2-cyclopentene-1-carbamate, and dl-p-methoxybenzyl-cis-5- (4-methylphthalimidoxy) -cyclopentene-1-carbamate.

20th

Preparation XVI

Preparation of dl-benzyl-cis-5- (phthalimidoxy) -2-cyclopentene-1-carbamate

I! If

An amount of 1.46 g (6.26 mmol) of trans-3-aminocyclo-20 ml fractions are collected. It shows up

pentene-N-benzyl urethane, 1.12 g (6.88 mmol) N-hydroxy- based on thin-layer chromatography that the fracture

Phthalimide and 1.8 g (6.88 mmol) of triphenylphosphine will contain the product for 13-20 years. Narrowing this frak-

dissolved in 25 ml of dry tetrahydrofuran. The solution was given 1.76 g of dl-benzyl-cis-5- (phthalimidoxy) -2-cyclo dropwise over a period of about 5 minutes with 1.32 g (7.6 mmol) of 60 pentene-1-carbamate in the form of a Oil, which

Diethylazodicarboxylat treated in 5 ml of tetrahydrofuran. let crystallized. Recrystallization from 50% vinegar

The initially red solution turns color about 10 minutes after ethyl acetate in hexane gives a melting point of

Start adding yellow. After 16 hours the reaction is 120-122 ° C.

solution concentrated in vacuo, leaving a pressure of The analytical values were the following:

less than 25 mm and a temperature of around 30 ° C 65

applies. Then chromatographed over 150 g NMR (CDCb, 5): 2.65-2.95 (m, CH2), 4.5-5.10 (m, 2H), 5.15

Silica gel. The column is washed with 40 ml of ethyl acetate (s, OCH2), 5.65-6.1 (m, CH = CH), 6.1-6.4 (NH), 7.37 (s,

benzene (20:80) and then (25:75) eluted. Fifty-ArH), 7.78 (s, 4, ArH).

646961

30th

TLC (silica gel 60): Rf = 0.65 using ethyl acetate / benzene (25:75) and Rf = 0.72 using acetone / methylene chloride (5:95) as the eluent.

Elemental analysis for C21H1SN2O5 gave the following values:

Calc .: C Found: C

66.65; 66.16;

H 4.79; H 4.84;

N N

7.41 7.32.

Using the same procedure, the d-benzyl-cis-5- (phthalimidoxy) -2-cyclopentene-1-carbamate is prepared by using the starting d material instead of the racemic mixture.

Preparation XVII

Preparation of dl-2,2,2-trichloroethyI-cis-5- (aminooxy) -2-cyclopentene carbamate

NH2NH2H2O

0 H il

NC0CH2CC13

h2no

0

H II

ncoch2cci3

I! ! d

An amount of 7.42 g (17.67 mmol) of dl-2,2,2-trichloroethyl-trans-5- (phthalimidoxy) -2-cyclopentene-l-carbamate was dissolved in 65 ml of tetrahydrofuran and 65 ml of ethyl alcohol . The solution is treated with 0.94 ml (0.97 g, 19.44 mmol) of hydrazine hydrate. A precipitate forms after 10 minutes. After 2 hours the reaction mixture is evaporated in vacuo using a pressure less than 25 mm and a temperature of approximately 30 ° C. The residue is treated with methylene chloride and filtered to separate the insoluble phthalhydrazide. The filtrate is concentrated in vacuo and 6.68 g of dl-2,2,2-trichloroethyl-cis-5- (aminooxy) -2-cyclopentene-l-carbamate remain in the form of an oil.

The analytical data were the following:

NMR (CDCb, 5): 2.35-2.65 (m, CH2), 4.1-4.95 (m, 2H) 4.78 (s, OCH2), 5.35-6.1 (m , 5, NH, NH2, (CH = CH).

ÌTLC (silica gel 60): Rf = 0.41 using methanol / benzene (5:95) and Rf = 0.41 using ethyl acetate / Skellysolve B (40:60) as the solvent.

Using the same procedure as in preparation XIII, but using different dl-halo alkyl-trans-5- (phthalimidoxy) -2-cyclopentene-l-carbamates instead of dl-2,2,2-trichloroethyl-trans-5- (Phthalimidoxy) -30 2-cyclopentene-1-carbamate, the corresponding halo-alkyl-cis-5- (aminooxy) -2-cyclopentene-1-carbamates are obtained.

Using the same procedure as in preparation XVII, but using dl- or d-benzyl-35 cis-5- (phthalimidoxy) -2-cyclopentene-l-carbamate, di or d-benzyl-cis-5- (aminooxy) -2-cyclopentene-l-carbamate obtained, the Rf value = 0.40 (mobile phase 50% ethyl acetate / hexane) and the Rf value of the starting material it is 0.71.

40

Preparation XVIII

Preparation of dl-2,2,2-trichloroethyl-cis - ({[(benzyloxy) -45 carbonyl] amino} oxy) -2-cyclopentene-1-carbamate

H2NO

0

H II

NCOCH2CCI3

O

CICOCH2C6H5

Illd cqh5 ch2 ocno

0

H II

NCOCH2CCI3

I I lc

The crude dl-2,2,2-trichloroethyl-cis-5-aminooxy-2-cyclopentene-1-carbamate (1.6 mmol), which was prepared in preparation XVII, is dissolved in 10 ml of pyridine and the solution is cooled in an ice bath and treated with 0.34 g (2.0 mmol) of benzyloxychloroformate, dissolved in 2 ml of methylene chloride. After 1.5 hours, the reaction mixture is poured into water and methylene chloride and treated with ammonium chloride until an acidic reaction is achieved. The methylene chloride layer is separated, dried over sodium sulfate and concentrated in vacuo, leaving 766 mg of an oil. The oil is eluted over 55 g of silica gel 60 chromatographies and with ethyl acetate / Skellysolve B in a ratio of 40:60. Then ml fractions are collected. Fractions 10-13 contain the product as it results from thin-layer chromatography and they are combined and concentrated in vacuo, leaving 535 g of a residue. After adding isopropyl ether to the residue and scratching, the material crystallizes and dl-2,2,2-trichloroethyl-cis - ({[(benzyl-65 oxy) -carbonyl] amino} oxy) -2-cyclopentene-1-carbamate is obtained with a melting point of 86-89 ° C.

The analytical results were as follows:

31

646 961

NMR (CDCb, 5): 2.2-2.9 (m, 2H), 4.2-5.0 (m, CHN, CHO) 4.67 (s, OCH2CCI3), 5.16 (s, OCH : 0), 5.4-6.2 (m, CH = CH), 7.37 (s, 0), 7.6-8.0 (m, NH), 8.47 (s, NH).

TLC (silica gel 60): Rf = 0.73 using ethyl acetate / Skellysolve B in the ratio 40:60 as the eluent.

Using the same procedure as in preparation XVIII, but using dl- or (IR, 5S) -benzyl-cis-5- (aminooxy) -2-cyclopentene-1-carbonate as a compound of the formula IIld and using 2 , 2,2-Trichloroethylchlor-formic acid ester instead of benz yloxychloroformate are correspondingly obtained dl- or d-benzyl-cis {[(2,2,2-trichloroethoxy) carbonyl] amino-oxy} -2-cyclopenten-1-carbamate.

The analytical data were the following:

NMR (CDCb): 7.30 (s, pH), 5.6-6.1 (m, 2H), 5.10 (5, OCH2), 4.4-5.0 (m, 2H), 4th , 72 (s, CCI3CH2), 2.4-2.8 (m, 2H).

TLC: Rf = 0.83 using 50% ethyl acetate / hexane.

10th

Preparation XIX

Preparation of dl-benzyl-cis - [(2-amino-3-cyclopenten-l-yl) oxy] carbamate

0

! Ih

CaHs CH2 0CN0

NC0CH2 CC13 H

ONCOCH2 CaH;

I I lc a) Zinc, ammonium chloride, methanol Process:

An amount of 21.32 g (50.4 mmol) of dl-2,2,2-trichloroethyl-cis - ({[(benzyloxy) carbonyl] amino} oxy) -2-cyclopenten-1-carbonai is obtained in 425 ml of methanol is dissolved and the solution is treated with 21.3 g (325 mmol) of zinc and 10.7 g (200 mmol) of ammonium chloride. The mixture thus obtained is stirred vigorously for 60 minutes and after this time it is evident from the thin layer chromatography that the reaction is complete. The reaction proceeds mildly exothermic and the temperature increased in the described batch scale from 25 ° C. at the start of the reaction to a maximum value of 35 ° C. The reaction mixture is then filtered and the filtrate is concentrated in vacuo. The filtered solids are washed with about 200 ml of 5% aqueous sodium bisulfate solution and the washing solutions are added to the residue of the methanol evaporation. The aqueous mixture thus obtained is extracted with methylene chloride and the methylene chloride layer is separated off. The aqueous layer is basified to pH 8 with concentrated ammonium hydroxide solution and then treated with 4 g of sodium cyanide and the solution obtained in this way is extracted three times with about 200 ml portions of methylene chloride. The combined methylene chloride solutions are dried over sodium sulfate and concentrated in vacuo, giving 9.6 g (77%) of benzyl cis - [(2-amino-3-cyclopenten-l-yl) oxy] carbamate in the form of an oil.

The analytical values were as follows:

NMR (CDCb, 5): 2.2-2.7 (m, 2H), 3.5-5.5 (m), 5.08 (s, OCH2), 5.5-5.9 (m, CH = CH), 7.28 (s, 5Ar-H).

TLC (silica gel 60): Rf = 0.42 using ammonium hydroxide-methanol-methylene chloride in the ratio 1:10:90 as the eluent.

Zinc complexes of the product dl-benzyl-cis - [(2-amino-3-cyclopenten-1-yl) oxy] carbamate are obtained from the reaction and are insoluble in methanol and most other organic solvents. They can be dissolved in aqueous acids.

The compound forms strong complexes with zinc at basic pH values. The product can only be extracted from the aqueous phase if the complexes are broken up. The ethylenediaminetetraacetic acid sodium

I I lb

25 salt is just as suitable for this but not as good as sodium cyanide.

b) zinc, ammonium chloride, methanol, sodium hydroxide workup

The zinc, ammonium chloride reduction in methanol is carried out as in step (a) above with an amount of 17.67 mmol dl-2,2,2-trichloroethyl-cis - ({[(benzyloxy) carbonyl] amino} oxy) -2- cyclopentene-1-carbamate. After extraction of the acidic aqueous solution to remove the dichloro by-product, the aqueous phase is made basic to pH 14 using sodium hydroxide and ice to keep the mixture cool. At pH 8-10, a voluminous precipitate of zinc hydroxide forms, but this dissolves again at higher pH values and the zinc salt of the desired product remains. This salt is insoluble in most solvents with the exception of aqueous acids. The precipitate thus obtained is filtered and dried. The zinc salt of dl-benzyl-cis - [(2-amino-3-cyclopenten-1-yl) oxy] carbamate is dissolved in the minimum possible amount of ammonium chloride necessary to effect the solution. It is then freeze-dried and the residue is used in preparation XXa.

c) zinc, methanesulfonic acid, methanol procedure 50 an amount of 10 g (23.6 mmol) of dl-2,2,2-trichloroethyl

cis - ({[(benzyloxy) carbonyl] amino} oxy) -2-cyclopentene-1-carbamate is dissolved in 200 ml of methanol. The solution is treated with 10 g (154 mmol) of zinc dust. 4 ml of 55 methanesulfonic acid are added to this mixture over 25 minutes with vigorous stirring. Thin layer chromatography shows that the reaction is completed in less than an hour. The reaction mixture is then filtered and the zinc solids are washed out with methanol. The filtrate and the combined washing solutions 60 are then concentrated in vacuo and the residue is used directly in preparation XX.

d) zinc, acetic acid, water procedure

An amount of 1.0 g (2.35 mmol) of dl-2,2,2-trichloroethyl-65 cis - ({[(benzyloxy) carbonyl] amino} oxy) cyclopentene-1-carbamate is dissolved in 10 ml of acetic acid water dissolved in a ratio of 9: 1 and the solution is added with a total amount of 1.0 g (15.4 mmol) of zinc in five equal portions

35

40

646961

32

45 minute intervals treated. After a total of 6 hours after the first addition, thin layer chromatography shows that the reaction is complete. The reaction mixture is then filtered and the filtrate is allowed to partition between 5% sodium bisulfate solution and methylene chloride. The aqueous layer is separated and basified to pH 9-10 with concentrated ammonium hydroxide solution using ice to keep the mixture cool. The aqueous solution thus obtained is extracted three times with methylene chloride. The combined methylene chloride solutions are over

Sodium sulfate dried and evaporated in vacuo, leaving 516 mg (88%) dl-benzyl-cis - [(2-amino-3-cyclopent-l-yl) oxy] carbamate in the form of an oil.

In the acetic acid experiments, ammonium hydroxide 5 appears to be suitable for breaking up the zinc salts. However, ammonium hydroxide is not sufficient for the products from procedures XIX (a) and XIX (c).

Preparation XX

io Preparation of dl-benzyl-cis- [2-phthalimide-3- (cyclopent-1-yl) oxy] carbamate (30)

(a) An amount of 11.6 g (46.6 mmol) of crude benzyl-cis - [(2-amino-3-cyclopenten-l-yl) oxy] carbamate from step 34, process (a) is dissolved in 250 ml Dissolved tetrahydrofuran. 11.5 g (58.3 mmol) of 2-methoxycarbonylbenzoyl chloride and 31 ml (23.5 g (232 mmol) of triethylamine are added. The reaction mixture is warmed to 50 ° C. for 72 hours and after this time the thin layer chromatography shows that the reaction is complete, the reaction mixture is then concentrated in vacuo, the residue is partitioned between ethyl acetate and ammonium chloride, the ethyl acetate layer is separated and washed with 5% sodium bisulfate solution and water and dried over magnesium sulfate , 9 g of a crude residue The residue is chromatographed over 2 kg of silica gel 60 by eluting with ethyl acetate / Skellysolve B in a ratio of 40:60. 400 ml fractions are collected after a flow of 21 has elapsed The product is in fractions 16-24 as indicated by thin layer chromatography can be seen. Concentration of these fractions gives 13.15 g (75%) of dl-cis- [2-phthalamid-3- (cyclopenten-yl) oxy] carbamate.

The analytical values were as follows:

NMR (CDCb S): 2.4-3.2 (m, CH2) 4.5-5.5 (m, 4), 5.00 (s, OCH2), 5.5-6.2, (m , CH = CH), 7.27 (s, 5 ArH), 7.5-8.0 (m, 4ArH).

TLC (silica gel 60): Rf = 0.48 when using ethyl acetate / Skellysolve B in a ratio of 40:60 as the eluent.

(b) Using the same procedure as in step (a) described above, 17.67 mmol crude zinc salts of dl-benzyl-cis - [(2-amino-3-cyclopenten-l-yl) oxy] carbamate processed and 3.23 g (48%) of 30-prey of cis- [2-phthalimid-3-cyclopenten-l-yl) oxy] carbamate are obtained.

(c) Using the same procedure as in steps (a) and (b) described above, using 23.6 mmol dl-benzyl-cis - [(2-35 amino-3-cyclopentene- l-yl) oxy] carbamate, 6.91 g (77%) yield of dl-cis - [(2-phthalimid-3-cyclopenten-l-yl) oxy] car-bamate.

The two stages XIX (c) and XX (c) are the preferred procedure for converting dl-2,2,2-trichlo-40 ethyl-cis - ({[(benzyloxy) carbonyl] amino} oxy) -2-cyclo -pentene-1-carbamate in dl-benzyl-cis- [2-phthalimide-3- (cyclopentene-1 -yl) oxy] carbamate.

In addition, dl- and d-cis- {2-phthalimide-3- [cyclopentene-1 - (2,2,2-trichloroethoxycarbonyl) aminooxy] car-45 bamate can be prepared using the procedure of Preparation XX, however with the exception that dl- or d-2,2,2-trichloroethyl-cis - [(2-amino-3-cyclopententen-l-yl) oxy] carbamate is used. These alternative starting materials can be prepared by dl- -50 or d-benzyl-cis - {[(2,2,2-trichloroethoxy) carbonyl] amino-oxy} -2-cyclopentene-l-carbamate (prepared in preparation XVIII ) with glacial acetic acid saturated with hydrobromic acid. Treated for 1 hour at room temperature. The reaction solution is concentrated in vacuo to give the product, which can be used directly in Preparation XX as above.

The analytical results were as follows:

NMR (CDCb): 7.5-8.0 (m, 4H), 8.45 (m, NH), 5.5-6.2 (m, «o 2H), 4.3-5.5 ( m, 4H), 4.69 (s, CCbCH :), 2.6-3.0 (m, 2H). TLC: Rf = 6.67 using ethyl acetate / hexane, 1: 1.

33

example 1

Preparation of (aS, 5S and aR, 5R) -2 - [(Benzyloxy) carbonyl] -3-oxo-a-phthalimide-5-isoxazolidine acetic acid

646 961

0

COoH +

ICOCH2C6Hs

CH2C6H5

I ic

An amount of 10.0 g (26.5 mmol) of dI-cis - [(2-phtha-limid-3-cyclopenten-l-yl) -oxy] carbamate of the formula purple is dissolved in 250 ml of acetone and 180 ml of water solved. The solution thus obtained is treated with 28 g (130 mmol) of sodium iodate and 100 mg of ruthenium chloride hydrate (1-3 H2O) in 5 ml of water. The reaction mixture is stirred vigorously for 1 hour. A slight exothermic reaction occurs after about the first 15 minutes, causing the temperature to rise from about room temperature (~ 23 ° C) to about 35 ° C. The reaction mixture is then evaporated at <30 mm and at a temperature of <30 ° C to remove in acetone. The aqueous residue is then partitioned between ethyl acetate and water which has been acidified with approximately 10 ml of 1M sulfuric acid. The aqueous layer is separated off and extracted twice with ethyl acetate. The combined ethyl acetate solutions are washed with saturated sodium chloride solution, dried over magnesium sulfate and concentrated in vacuo, using a pressure of less than 30 mm and a temperature of less than 30 ° C., leaving a glassy foam. The residue is chromatographed on 1 kg of CC-4 silica gel by eluting with a linear gradient which is prepared using 51 ethyl acetate / Skellysolve B (40:60) and 51 ethyl acetate / Skellysolve B (80:20). Fractions (400 ml) 16-19 gave a yield of 7.48 g of an essentially pure (cc5.5S and 2R, 5R) -2 - [(benzyloxy) carbonyl] -3-oxo-a-phthalimide-5 -isoxazoline-acetic acid with a melting point of 113-116 ° C (from ethanol).

The analysis results were as follows:

NM R (dô-acetone 8): 3.16 (d, = 7.5, CH2), 5.25 (s, OCH2), 5.1 -5.7 (m, 2), 7.39 (s , 5), 7.92 (s, 4).

The elementary analysis calculated for C21H10N2O8 gave the following values:

Calc .: C 59.43; H 3.80; N 6.60 Found: C 58.79; H 3.80; N 6.53.

Fraction 14 gave 0.93 g2- [2 (benzyloxy) carbonyl] -3-oxo-4-phthalimide-5-isoxazolidine-acetic acid with a melting point of 160-162 ° C. (from ethyl acetate / hexane, 1: 1) .

The analytical values were as follows:

NMR (dó-acetone 8): 3.06 (d, J = 5.5, CH2), 5.38 (s, OCH2), 5.1-5.8 (m, 2), 7.45 (m , 5), 7.88 (s, 4).

The elementary analysis calculated for C2iHi6N202 • H2O gave 25 the following values:

Calculated: C 57.01; H 4.10; N 6.33 Found: C 57.24; H 3.98; N 6.45.

30 A mixture of the two acids was found in fraction 15.

Using the same procedure as in Example 1, but using dl- or d-cis- (2-phthalimide) -3- [cyclopentene-1 (2,2,2-trichloroethoxy-35 carbonyl) aminooxy] carbamate man (a5,5S and aR, 5R) -2 - [(2,2,2-trichloroethoxy) carbonyl] -3-oxo-a-phtha-limid-5-isoxazolidine-acetic acid in 57% yield with a melting point of 208-210 ° C.

The analytical results were as follows:

40

NMR (acetone-da): 3.27 (d, J = 7.5 Hz, CH2), 4.92 (s, CCb-CH2), 5.1-5.7 (m, 2H), 7.91 (s, 4H).

TLC: RF = 0.41 (A IX).

45 The solvent called “AIX” used to perform thin layer chromatography is the organic phase that is separated after the following components have been mixed:

50 35% ethyl acetate 20% 2,2,4-trimethylpentane 40% water 5% acetic acid

55 The elementary analysis calculated for C16H11CI3N2O8 gave the following values:

Calculated: C 41.27; H 2.38; N 6.02 Found: C 41.33; H 2.36; N 6.04.

60

The isomers (4S, 5S and 4R, 5R) -2 - [(2,2,2-trichloroethoxy ~ carbonyl] -3-oxo-4-phthalimide-5-isoxazolidine-acetic acids are also formed in 22% yield The analytical values for this were:

65

NMR (acetone-ds): 3.10 (d, J = 5.5 Hj, 2H), 5.08 (5, CCb, CH2), 5.1 -5.8 (m, 2H), 7.90 (s, 4H).

TLC: Rf = 6.52.

646 961

34

Example 2

Preparation of (ct5,5S and cxR, 5R) -3-oxo-a-phthalimide-5-isoxazolidine acetic acid

0 ii c-

0 /

ch2 I

CeHs i c

I Ib

An amount of 7.48 g (17.6 mmol) of (ccS, 5S and aR, 5R) -2 - [(Benzyloxy) carbonyl] -3-oxo-cc-phthalimide-5-isoxazolidine-acetic acid is dissolved in 150 ml of ethyl acetate and 75 ml of 95% ethanol dissolved. The solution is treated with 1.5 g of palladium black and hydrogenated at 25 ° C. and 1 atmosphere of pressure. The reaction was stopped after 165 minutes and filtered and the filtrate was used directly for the preparation of (aS, 5S and aR, 5R) -3-oxo-cc-phthalimid-5-isoxazolidine acetic acid.

In a further approach, using the same procedure as described above, the filtrate was evaporated in vacuo to give (aS, 5S and aR, 5R) -3-oxo-a-phthalimide-5-isoxazolidine acetic acid as a residue.

The analytical values were as follows:

NMR (CD3OD, 8): 2.92 (partially split d, CH2), 5.1-5.7 (m, 2), 7.85 (s, 4 ArH).

TLC (silica gel 60): Rf = 0.36 using the supernatant phase of ethyl acetate-acetic acid-cyclo-hexane-water in the ratio (9: 2: 5: 10) as the eluent. In the same system, the starting material shows an Rf value of 0.46.

Using the same procedure as in Example 2 using the dl or d isomers of the formula VIHd, the dl or d isotricholomic acid phthalimides of the formula Vile are obtained in the form of a white co2h

Foam with an Rf value of 0.54 in thin layer chromatography (Rf value of the starting material = 20 0.64).

NMR (acetone-do): 9.1 (s, CChH), 7.83 (s, 4H), 5.0-5.8 (m, 2H), 2.95 (m, 2H).

Alternatively, the benzyloxycarbonyl group can be removed from the compound of formula Ile to produce the compound of formula IIb (or convert Vlld to Vile) using acid. For example, an amount of 425 mg (1 mmol) of the compound of formula 30 IIb is dissolved in 5 ml of glacial acetic acid and a nitrogen atmosphere is maintained and dry hydrobromic acid is introduced into the reaction solution for 10 minutes. After stirring for 4 hours at room temperature, the solution is concentrated and chromato-35 is graphed over 50 g of CC-4 silica gel with 70% ethyl acetate / hexane. This procedure yields 170 mg (59%) of a product that is identical to the product of hydrogenative cleavage (based on nuclear magnetic resonance spectrometry and thin layer chromatography). Other acids and 40 solvents such as e.g. HBr / CHîNCh, HBR / CHaCbund trifluoroacetic acid can also be used.

Example 3

Preparation of the benzhydryl esters of (aS, 5S and <xR, 5R) -3-45 oxo-a-phthalimid-5-isoxazolidin-acetic acid.

2nd

I Ib

A quantity of 5.2 g (26.5 mmol) of benzophenone hydrazone was dissolved in 125 ml of ethyl ether. The solution is treated with 10 g of sodium sulfate, 11.1 g (51 mmol) of yellow mercury oxide and 0.2 ml of saturated potassium hydroxide solution in ethyl alcohol. The reaction mixture is stirred for 60 minutes and the deep burgundy obtained in this way

The mixture is filtered. The solution thus obtained is added directly to the reaction solution in which 17.6 mmol (aS, 5S 65 and aR, 5R) -3-oxo-a-phthalimide-5-isoxazolidine-acetic acid had been hydrogenated. Thin layer chromatography shows that the reaction is complete in less than 1 hour. The reaction solution is then extracted with

35

646961

appropriate amounts of 3N hydrochloric acid while stirring vigorously to destroy the excess of the burgundy colored diazo compound. The reaction mixture is then concentrated in vacuo, using a pressure of less than 30 mm and a temperature of less than 30 ° C. and chromatographed over 500 g of CC-4 silica gel and eluted with ethyl acetate-toluene in a ratio of 40:60. Two hundred ml fractions were collected. The benzhydryl esters of (aS, 5S- and aR, 5R) -3-oxophthalimide-5-isoxazolidine-acetic acids were in fractions 10-14 and a yield of 3.05 g (38%) was obtained.

The analytical data were the following:

NMR (CDCb Ô): 2.75 (d, J = 11.5, CHa), 5.15-5.7 (m, 2), 6.93 (s, CH (C6Hs) 2), 7.18 (s, CôHs), 7.28 (s, CôHs), 7.5-8.0 (m, 4, ArH).

TLC (silica gel 60): Rf = 0.42 using ethyl acetate / toluene in a 40:60 ratio as the eluent.

The isomeric product of the compound VIII (see Example 1) shows an Rf value of 0.59 in the same system.

Using a procedure similar to that of Preparation XX, but replacing RN2 with diphenyldiazomethane, where R can represent a methyl group, an ethyl group, a benzyl group and the like, the corresponding esters of formula IIb are obtained. With these more reactive alkyldiazo compounds

It is more effective to use the formula IIb acid in a non-protic solvent, e.g. Tetrahydrofuran, instead of treating ethanol.

Alternatively, the esters of formula IIb can be prepared by alkylating the acid in base with a reactive alkylating agent. For example, an amount of 87 mg (0.3 mmol) of the compound of the formula IIb in 3 ml of dry acetonitrile under nitrogen can be treated with 50 ml of diisopropylethylamine and 60 mg of p-methoxybenzyl bromide. After 16 hours, the solution is partitioned between ethyl acetate and water and the organic phase is separated off, dried over sodium sulfate and concentrated. Preparative thin layer chromatography using 50% ethyl acetate-15 ester / hexane gives 55 mg (45%) of the p-methoxybenzyl ester.

The analytical data were the following:

NMR (CDCb): 7.81 (m, 4H), 7.05 (A2B2.4H), 5.1-5.7 (2H), 5.13 (s, 2H), 3.76 (s, 3H) , CHsO), 2.81 (d, J = 7.5 Hs, 2H).

20th

Using the procedures described above, but using the dl- or d-isotricho-lomic acid phthalimides of the formula Vile instead of the compounds of the formula Ile, the dl- and d-iso-25 tricholomphthalimide esters of the formula VHIb are obtained in each case.

Preparation XXI

Preparation of dl-trans-3-phthalimide-4-fluoromethanesulfonyloxycyclopentene

0 = £ = o

An amount of 5 g (21.8 mmol) of 4-hydroxy-4-phthalimide cyclopentene and 1.72 g (21.8 mmol) of pyridine are dissolved in 6 ml of methylene chloride and the solution is added dropwise to an ice-cold solution over 40 minutes 4.6 g (21.8 mmol) of trifluoromethanesulfonic anhydride in 15 ml of methylene chloride are added under a nitrogen atmosphere with stirring. The solution is stirred for a further 15 minutes and then washed with water and dried and finally concentrated with methylene chloride solution, leaving dl-trans-3-phthalimide-4-trifluoromethanesulfo-45 nyloxycyclopentene.

Preparation XXII

Preparation of dl-BenzyI-cis - [(2-phthalimid-3-cyclo-50 penten-yl) oxy] carbamate.

0

1

ch2

i <°>

An amount of 21.8 mmol of dl-trans-3-phthalimide-4-tri-fluoromethanesulfonyloxycyclopentene is dissolved in 50 ml of methylene chloride and the solution is treated with 21.8 mmol of calcium salt of N-hydroxy-benzyl urethane. After stirring for 24 hours under a nitrogen atmosphere, the reaction mixture is extracted with 5% aqueous sodium 65 um bicarbonate solution and water and dried. Distilling off the methylene chloride gives a residue, which is chromatographed on silica gel, using 40:60 with ethyl acetate / Skellysolve B.

646961

36

eluted. After concentration of the fractions which are due to the, d-benzyl-cis - [(2-phthalimide-3-cyclopentene thin layer chromatography gives the desired product entyl) oxy] carbamate.

Production of optically active isomers

Preparation XXIII

Separation of dl-trans-3-amino-4-hydroxycyclopentene (5): into its optical antipodes: preparation of (+) - deoxycholate 6b and (-) - tartrate 6c.

1. Deoxycholic acid

2. Amberlite IRA-400

3. L (+) tartaric acid

IVb

1) An amount of 49.5 g (0.50 mol) of dl-trans-3-amino-hydroxycyclopentene dissolved in about 100-200 ml of methanol and 98 g (0.25 mol) of deoxycholic acid dissolved in about 200 to 300 ml Methanol is mixed and allowed to crystallize for several hours. The crystals of trans-3-amino-4-hydroxycyclopentene are collected by filtration and washed three times with small portions of methanol. This gives 105 g of trans-3-amino-4-hydroxycy-clopenteneoxycholate in the form of light beige crystals with a melting point of 195-197 ° C (decomposition). The mother liquors are saved for step 2.

2) A quantity of 500 ml of Amberlite IRA-400 (chloride form) is poured into a 600 ml glass sinter funnel and so it is washed six times with 200 ml portions in sodium hydroxide three times with water (3x200 ml) with 200 ml portions of water , three times with 200 ml of methanol. A quantity of 250 ml of this resin in the hydroxyl form and the mother liquors from step 1 above are mixed and the mixture is stirred for 2 hours under a nitrogen atmosphere. The mixture is then applied to the top of a chromatography column packed with a further 250 ml of resin in methanol. The column is eluted with methanol by collecting 250 ml fractions. Product 60 is present in fraction # 106 based on the results of thin layer chromatography. Evaporation of these fractions gives 32 g (3R., 4R) -3-amino-4-hydroxycyclopentene.

Thin layer chromatography on silica gel 60 gives an Rf value of 0.24 using a solvent 65 from ammonium hydroxide-methanol-methylene chloride in a ratio of 1:20:80. The detection was carried out by spraying with potassium permanganate.

3) The residue from step 2) above is dissolved in 300 ml of 95% ethanol and the solution is dissolved in a solution of 37.5 g (0.25 mol) of L - (+) - tartaric acid in 300 ml of 95 % ethanol, mixed. The mixed solutions are inoculated with salt which has already been separated on the optical antipodes. After 2-3 hours, the crystals are collected by filtration and dried. 54.4 g of crystals with a melting point of 84 to 86 ° C. are obtained in this way. The crystals are recrystallized from 700 ml of 95% ethanol. In this way, 48.5 g of crystalline (3R, 4R) -3-amino-4-hydroxycy-clopentene L (+) - tartaric acid salt with a melting point of 86.5-88.5 ° C and an optical rotation of [ a] ^ 8 = 37.8 ° (c = 3.20 in methanol).

4) A quantity of 200 ml of Amberlite IRA-400 (hydroxyl form) and 300 ml of methanol were added to 25 g of separated tartaric acid salt from step 3). The mixture is stirred under a nitrogen atmosphere for 1 hour. A further 100 ml of Amberlite IRA-400 (hydroxyl form) are then introduced into a chromatography column and the above mixture of the tartaric acid salt, amberlite resin and supernatant methanol solution were applied to the top. The methanol is drained from the column and fresh methanol is passed through until a total of 1500 ml of methanol is collected. The methanol solution is then evaporated in vacuo to give 10.18 g of crystalline residue, namely (3R, 4R) -3-amino-4-hydroxycyclopentene with a melting point of 67-75 ° C and an optical rotation of [a]; 8 = -139 ° (c = 1.0, MeOH) remain.

40

45

37 646961

Preparation XXIV

Preparation of (1R, 5R) -2,2,2-trichloroethyl-5-hydroxy-2-cyclopentene-1-carbamate

(a)

(+)

o ii

C1COCH2CQ3 NazCOj

(+)

0 H II

NCOCM:

CCI 3

OH (+) IVb

An amount of 10.18 g (103 mmol) of crude (3R, 4R) -3-amino-4-hydroxycyclopentene (prepared as in preparation XXIII, step 4) is dissolved in 75 ml of water and the mixture is mixed with 10.6 g (100 mmol) sodium carbonate treated. The mixture is then cooled to less than 10 ° C with an ice bath and treated dropwise with vigorous stirring with 24 g (110 mmol) of trichloroethyl chloroformate over a period of 30 minutes. After 3 hours the reaction mixture is filtered and the collected solids are washed thoroughly with water. Drying gives 22.43 g (1 R, 5 R) -2,2,2-trichloroethyl-5-hydroxy-2-cyclopenten-1-car-

HO (+) 'IVa bamat in the form of a crystalline solid with a melting point of 82-84 ° C. Recrystallization of a small sample from isopropyl ether Skellysolve B gives a material with a melting point of 87-87.5 ° C and an optical rotation of [a] -92 ° (C = 0.14, MeOH).

Thin layer chromatography on silica gel 60 gives an Rf value of 0.5 in acetone-methylene chloride 10:90 as the eluent.

2s preparation XXV

Preparation of (IR, 5S) -2,2,2-trichloroethyl-5- (phthalimide oxy) -2-cyclopentene-1-carbamate

NCOCHaCC

(+) IVa

(CsHs ^ P

(= NC02C2H5) 2

THF

0

Njlll ncoch3cci3

(+) II le

An amount of 21 g (76.5 mmol) (IR, 5R) -2,2,2-trichloroethyl-5-hydroxy-2-cyclopentene-1-carbamate, 13.76 g (84.4 mmol) N Hydroxyphthalimide and 22.1 g (84.4 mmol) of tri-phenylphosphine are dissolved in 400 ml of dry tetrahydrofuran. 16.12 g (92.6 mmol) of diethyl azodicar boxylate is added dropwise to the solution with stirring over a period of 15 minutes, while keeping the temperature below 35 ° C. using an ice bath. The reaction is then stirred at 25 ° C. for 1 hour and then concentrated in vacuo at a pressure of less than 25 mm and at a temperature of approximately 30 ° C. The residue is treated with approximately 100 ml of ethyl acetate-Skellysolve B in a ratio of 10:60 and the separated triphenylphosphine oxide is separated off by filtration after about 15 minutes. The residue is applied to the top of a column packed with 2 kg of silica gel 60, which is eluted with 3 liters of ethyl acetate / Skelly-60 solve B in a ratio of 40:60 and then in a ratio of 50:50. Fractions of 300 ml each are collected. Fractions 19-27 contain the pure (IR, 5S) -2,2,2-trichloroethyl-cis-5- (phthalimidoxy) -2-cyclopentene-1-carbamate, as is shown by the thin-layer chromatography-65 phie. Concentration gives 26.9 g (IR, 5S) -2,2,2-trichloroethyl-5- (phthalimidoxy) -2-cyclopenten-1-carbamate with a melting point of 117-118 ° C and an optical rotation of [ a] o = -26 ° (C = 0.56, MeOH).

646 961 38

Preparation XXVI

Preparation of (IR, 5S) -2,2,2-trichloroethyl-5- (aminooxy) -2-cyclopentene-1-carbamate

NH2NH2H2O

0

nc0ch2cc13

HpNO

0

h II

NCOCH2CCI3

I! Id

An amount of 24.9 g (17.67 mmol) (IR, 5S) -2,2,2-trichloroethyl-5- (phthalimidoxy) -2-cyclopenten-1-carbamate is dissolved in 230 ml of tetrahydrofuran and 230 ml Dissolved ethyl alcohol. The solution is treated with 3.3 ml (3.4 g 68 mmol) hydrazine hydrate. A precipitate forms after 10 minutes. After 2 hours the reaction mixture is evaporated in vacuo using a pressure of less than 25 mm and a temperature of about 30 ° C. The residue is treated with methylene chloride and filtered to remove the insoluble phthalhydrazide. The filtrate is concentrated in vacuo, leaving 23.3 g of a semi-solid. This material is treated with 100 ml of ethyl acetate and filtered. The filtered solids are washed with 100 ml of methylene chloride. The combined filtrates are dried in a vacuum.

kene evaporated to give 18.51 g (IR, 5S) -2,2,2-trichloroethyl-5- (aminooxy) -2-cyclopentene-1-carbamate in the form of an oil.

Thin layer chromatography on silica gel 60 gives an Rf of 0.41 using a mobile solvent of methanol / benzoyl in a ratio of 5:95 and an Rf of 0.41 using a mobile solvent of ethyl acetate / Skellysolve B in a ratio of 40 : 60.

Using the same procedure as in Example 23, but using the appropriate haloalkyl- (IR, 5S) -5- (phthalimidoxy) -2-cyclopenten-1-carbamate instead of (IR, 5S) -2,2, 2-trichloroethyl-5- (phthalimide-oxy) -2-cyclopentene-1-carbamate, the corresponding 30 haloalkyl-5- (aminooxy) -2-cyclopentene-1-carbamates are obtained.

20th

25th

Preparation XXVII

Preparation of (IR, 5S) -2,2,2-trichloroethyl-5 - ({[(benzyloxy) carbonyl] amino) oxy) -2-cyclopentene-1-carbamate

0

Hll nc0ch2ccl3

O

C1COCH2C6H5

0 Hll nc0ch2cc13

h2no c6h5ch2ocno

Id

IIIc

An amount of 18.5 g of crude (IR, 5S) -2,2,2-trichloroethyl-5-aminooxy-2-cyclopentene-1-carbamate, prepared as in preparation XVII, is dissolved in 175 ml of pyridine and the solution becomes cooled with an ice bath and treated with 11.6 g (67.7 mmol) of benzyloxychloroformate dissolved in 15 ml of methylene chloride. After 1½ hours the reaction mixture is treated with 10 ml and stirred for 10 minutes and the reaction mixture is then poured into water and methylene chloride and treated with ammonium chloride until an acidic reaction is obtained. The methylene chloride layer is separated, dried over sodium sulfate and concentrated in vacuo to give 22.6 g of an oil. This oil is recrystallized from ether, giving 5.2 g of (IR, 5S) -2,2,2-trichloroethyl ({[(benzyloxy) carbonyl] ami-no} oxy) -2-cyclopentene-1-carbamate with a Receives melting point of 89.5-90.5 ° C. A second crystal fraction of 9.17 g has a melting point of 89-90 ° C. and a third crystal fraction of 1.54 g has a melting point of 80-85 ° C. and is obtained by crystallizing the mother liquors of the first crystal fraction from isopropyl ether. The optical rotation was [a] d = -33 ° (C = 0.02, MeOH).

Thin layer chromatography on silica gel 60 gave an Rf value of 0.73 using ethyl acetate / Skellysolve B in a ratio of 40:60 as the eluent.

Preparation XXVIII

Preparation of (IR, 5S) -2,2,2-trichloroethyl-5 - ({[(t-butyl-oxy) carbonyl] amino} oxy) -2-cyclopentene-1-carbamate ss. A quantity of 7.8 g crude (IR, 5S) -2,2,2-trichloroethyl-5- (aminooxy) -2-cyclopentene-1-carbamate is dissolved in 25 ml of tetrahydrofuran and 6.15 g (25 mmol) of 2- (tert- Butoxycarbonyloxyimino) -2-phenylacetonitrile added and the mixture is stirred for 22 hours at room temperature and finally 20 hours at 50 ° C and 72 hours at room temperature and then evaporated in vacuo. The residue is chromatographed over 900 g of silica gel and eluted with ethyl acetate / toluene in a ratio of 15:85. Three hundred ml fractions of 65 are collected. The product is present in fractions 10-12. Evaporation and recrystallization of the residue from Skellysolve B gives (IR, 5S) -2,2,2-trichloroethyl-5 - ({[(t-butyloxy) carbonyl] amino} oxy) -2-cyclopentene-

39

646 961

1-carbamate with a melting point of 83-84 ° C in the form of preparation XXIX

of a white solid. Preparation of (1 S, 2R) -Benzyl - [(2-amino-3-cyclopenten-

NMR (CDCb, 8): 1.45 (s, 9), 2.45 = 2.7 (m, 2), 4.3-4.9 (m, l-yl) oxy] carbamate 2), 4 , 75 (s, 2), 5.6-6.1 (m, 3H), 7.7-7.8 (b, l).

0

II "

cahsch20cn0

NC0CH2 cc13 h

I c

HI

oncoch2c6h5

IIb

An amount of 7.12 g (16.8 mmol) of (IR, 5S) -2,2,2-trichloroethyl-5 - ({[(benzyloxy) carbonyl] amino} oxy) -2-cyclopentene-1 carbamate dissolved in 75 ml of methanol. The solution is treated with 7 g (108 mmol) of zinc dust. 2.1 ml of methanesulfonic acid are added to this mixture over the course of 10 minutes with vigorous stirring. Thin layer chromatography shows that the reaction is complete in less than 90 minutes. The reaction mixture is then filtered and the zinc solids are washed with methanol. The filtrate and the combined washing solutions are evaporated in vacuo to give 5.8 g of crude (1S, 2R) -benzyl - [(2-amino-3-cyclopenten-1-yl) oxy] carbamate, which is obtained directly in the preparation XXVII continues to be used.

Using the same procedure as in 20 preparation XXIX, but using (IR, 5S) -2,2,2-trichloroethyl-5 - ({[(t-butyloxy) carbonyl] amino} oxy) -2-cyclo- pentene-1-carbamate instead of (IR, 5S) -2,2,2-trichloroethyl-5 - ({[(benzyloxy) carbonyl] amino} oxy) -2-cyclopentene-1-car-bamate (IS, 2R ) -t-Butyl-5 - [(2-amino-3-cyclopenten-l-25 yl) oxy] carbamate.

Preparation XXX

Preparation of (IS, 2R) -benzyl- [2-phthalimide-3- (cyclo-30-penten-l-yl) oxy] carbamate

CqH5CH20CN0

"H

c6h5ch2ocno

I! b

I IIa

An amount of 5.8 (16.8 mmol) crude (IR, 5R) -benzyl- [2-amino-3-cyclopenten-l-yl) oxy] carbamate from preparation XXVII is dissolved in 90 ml of tetrahydrofuran. 4.95 g (25 mmol) of 2-methoxycarbonylbenzoyl chloride and 10 ml (7.3 g, 72 mol mol) of triethylamine are added to this solution. The reaction mixture is heated to 50 ° C for 72 hours and after this time thin layer chromatography shows that the reaction is complete. The reaction mixture is then evaporated in vacuo. The reaction mixture is then allowed to distribute between ethyl acetate and ammonium chloride. The ethyl acetate layer is separated off and washed with 5% sodium bisulfate solution and water and dried over magnesium sulfate. Evaporation of the ethyl acetate phase in vacuo gives 7.4 g of a crude residue. This is chromatographed over 600 g of silica gel 60 and eluted with ethyl acetate / Skellysolve B in a ratio of 40: 60.50 ml fractions are collected. The proso product is in fractions 16-24, as can be seen from the thin layer chromatography. Concentration of these fractions gives 5.2 g (75%) (IS, 2R) -benzyl- [2-phthalamid-3- (cyclopenten-yl) oxy] carbamate in the form of a crystalline solid with a melting point of 55 89-92 ° C and an optical rotation of [ci] d = -143 ° (C = 0.7, MeOH).

Thin layer chromatography on silica gel 60 gives an Rf value of 0.48 using ethyl acetate / Skellysolve B in a ratio of 40:60 as the eluent.

646961

40

Example 4

Preparation of (aS, 5S) -2 - [(Benzyloxy) carbonyl] -3-oxo-a-phthalimide-5-isoxazolidine-acetic acid co2h cHgCsHs

W

Illa Ile Vlld

An amount of 10.0 g (26.5 mmol) of (2R, 3S) -benzyl- [2- 2 - [(benzyloxy) carbonyl] -3-oxo-a-phthalimide-5-isoxazoline-phthalimide-3-cyclopentene -1 -yl) oxy] carbamate of the formula acetic acid with a melting point of 113 to 116 ° C and purple is dissolved in 250 ml of acetone and 180 ml of water. Such an optical rotation of [cc] d = -57 ° (C = 2.0 in MeOH). solution obtained is treated with 28 g (130 mmol) of sodium iodate. Fraction 14 gives 0.93 g (45.5S) -2 - [(benzyloxy) -car-

and 100 mg of ruthenium chloride hydrate (1-3 H2O) in 5 ml of 2s bonyl] -3-oxo-4-phthalimide-5-isoxazolidine acetic acid treated with water. The reaction mixture is stirred vigorously for a melting point of 160-162 ° C. (from acetic acid ethyl for one hour. The reaction mixture is ester / hexane 1: 1).

then at a pressure of less than 30 mm and at a A mixture of the two acids is evaporated in fraction 15 temperature of less than 30 ° C to the bound.

Remove acetone. The aqueous residue is then allowed to partition between ethyl acetate and water using the same procedure as in Example 4, but using (2R, 3S) -t-butyl- [2-which is acidified with about 10 ml of 1M sulfuric acid phthalimide- 3-cyclopenten-1-yl) oxy] carbamate instead of. The aqueous layer is separated off and (2R, 3S) -benzyl- [2-phthalimid-3-cyclopenten-l-yl) oxy] car-extracted again twice with ethyl acetate. The verbamate becomes (a5,5S) -2 - [(t-butyloxy) carbonyl] -3-oxo-a-unified ethyl acetate solutions with saturated 35 phthalimide-5-isoxazolidine-acetic acid and (4S, 5S) -2- [(t-sodium chloride solution washed, over magnesium sulfate butyloxy) carbonyl] -3-oxo-4-phthalimide-5-isoxazolidine-acetic acid-dried and evaporated in vacuo to give less acid.

than 30 mm pressure and less than 30 ° C and the analytical data were as follows:

got a glassy foam. The residue is placed on the top of a column of 40 NMR (CDCb, 5) packed with 1 kg of CC-4 silica gel: 1.35 (s, 9), 2.7-3.0 (m, 2), 4.55- 5.0 (m, 1), applied together with 80 g of CC-4 silica gel, to 5.15-5.55 (m, 1), 5.55-6.25 (m, 2), 7 , 2-7.4 (1), 7.6-8.0 (m, 4). which it was stored by evaporating the methylene chloride and then eluting. Four Hundred Example 5

dert ml fractions are collected. Evaporation of the production of (a5,5S) -3-oxo-a-phthalimide-5-isoxazo-

Fractions 16-19 give 7.48 g essentially pure (aS, 5S) - 45 lidin-acetic acid

"An amount of 7.48 g (17.6 mmol) (aS, 5S) -2 - [(benzyl- whereby 3-oxo-a-phthalimide-5-isoxazolidine-acetic acid oxy) carbonyl] -3-oxo- a-phthalimide-5-isoxazolidine acetic acid is obtained as a residue.

is in 150 ml of ethyl acetate and 75 ml of 95%

Dissolved ethanol. The solution is dissolved with 1.5 g of palladium black. The analytical data were the following:

treated and hydrogenated at 25 ° C under 1 atmosphere

Print. The reaction is stopped after 165 minutes, NMR (CD3OD, 5): 2.9 2 (partially split d, CH2), 5.1-5.7

it is filtered and the filtrate is evaporated in vacuo, (m, 2), 7.85 (s, 4 ArH).

41

646 961

Thin layer chromatography on silica gel 60 gave an Rf of 0.36 in the upper phase of ethyl acetate / acetic acid / cyclohexane / water in a ratio of 9: 2: 5: 10. In the same system, the starting material shows an Rf value of 0.54 and the compound Vlld shows an Rf value of 0.64.

Example 6

Preparation of the Benzhydrylester of (a5,5S) -3-Oxo-a-

an Rf of 0.46 and the hydrogenated product of VldS phthalimide-5-isoxazolidine-acetic acid

0oH

1 Ib

HNn 1 .C02CH (CeHs) 2

I i a

An amount of 2.8 g (14.3 mmol) of benzophenone hydrazone was dissolved in 80 ml of ethyl ether. The solution was treated with 10 g of sodium sulfate, 6.02 g (28 mmol) of yellow mercury oxide and 0.2 ml of a saturated potassium hydroxide solution in ethyl alcohol. The reaction mixture is stirred for 60 minutes and the deep burgundy mixture thus obtained was filtered. The solution thus obtained was added directly to the reaction solution from 8.45 mmol hydrogenation solution in which (a5.5S) -3-oxo-a-phthalimide-5-isoxazolidine-acetic acid was prepared. After one hour the reaction mixture is treated with sufficient 3N hydrochloric acid while stirring vigorously to destroy the excess of the burgundy colored diazo compounds. The reaction mixture is then evaporated in vacuo at a pressure of less than 30 mm and a temperature of less than 30 ° C. and chromatographed over 300 g of CC-4 silica gel by using

Ethyl acetate / toluene in the ratio 40:60 eluted. Thirty 5 ml fractions are collected. The benzhydryl ester of (aS, 5S) -3-oxo-a-phthalimid-5-isoxazolidin ~ acetic acid is in fractions 10-14 and shows an optical rotation of [ci] d = + 26 ° (C = 0.70). The analytical data were the following:

30th

NMR (CDCb, 5): 2.75 (d, J = 11.5, CH?), 5.16-5.7 (m, 2), 6.93 (s, CH (CóHs », 7.18 (s, CóHs), 7.28 (s, CsHs), 7.5-8.0 (m, 4 ArH).

TLC (silica gel 60): Rf = 0.42 using ethyl acetate / toluene in a ratio of 40:60 as eluent. The iso product of the formula VIIIb showed an Rf value of 0.59 in the same system.

Preparation XXXI 40 Production of tricholomic acid co2

*

co2 '

20 ml of 2N sodium hydroxide solution are added to 1.20 g (6.75 mmol) of (aS, 5S) -cc-amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid (AT-125) and the mixture is stirred at 25 ° C for 36 hours. The solution is adjusted to about pH 3.5 using 6N hydrochloric acid (dropwise addition) and then kept at -10 ° C (in a refrigerator) overnight. The crystals are collected by filtration, washed twice with cold water and then dried, and 720 mg of tricholomic acid are obtained. The first filtrate is lyophilized and the powder thus obtained is taken up in 4 ml of water, filtered and washed with 2 portions of 4 ml of cold water and dried, and a further 160 mg of tricholomic acid are obtained (total 80% yield).

The analytical data were the following:

60 NMR (D2O): 5.65-5.35 (1 H, m, ring CH), 4.27 (1 H, d, J = 3.5 Hs, cc-H), 3.69 (2H, d, J = 9.5 H3, CH2),

TLC: Rf = 0.31 (detection reagent ninhydrin), eluent: 60% methyl ethyl ketone, 20% acetone, 15% water and 5% acetic acid.

65 C NMR (D2O): 154.6 (CONH), 79.4 (CHO), 56.8 (<x-C), 36, S (CH2)

646961

42

Example 7

Preparation of the methyl ester of phthalyl tricholomic acid

C02

0

co2CH3 I I a2

methane. Chromatography on silica gel with 75% E / H gave 160 mg.

The analytical values were as follows:

A sample of 400 mg (2.5 mmol) of tricholomic acid,

720 mg of sodium carbonate, 6 ml of water and 1.2 g of N-carb ethoxyphthalimide are mixed together in three portions during the first 2 hours. After acidification to pH 3 with 3N hydrochloric acid, the reaction mixture is 20 NMR (CDCb): 5.65, 5.3 (1H, m, CHO), 5.3-5.1 (1H, m, mixture with 2x25 ml of ethyl acetate The ver aH), 3.06 (2H, d, CH2, PHTH unchanged).

united organic phases are with saturated cooking TLC: (AIX) Rf = 0.33 (50% E / H), Rf = 0.15; (AT-125 salt solution washed, dried over sodium sulfate and in the derivative) Rf = 0.42.

Vacuum concentrated. Chromatography on CC-4 silica gel with 50-75% E / H gradient gives phthalyl tricholomic acid. 2s analysis:

The analytical values are as follows:

Calculated: C 55.26; H 3.98; N 9.21 NMR (methanol d4): 7.86 (4H, s, 0), 5.65-5.0 (2H, m, CHO), found: C 55.20; H 4.60; N 8.43.

a-H), 2.92 (2H, d, CH2).

TLC: (AIX) Rf = 0.15 (detection was carried out using Ultravio-30 mass spectrometry: M + m / e 304 (3%), 272 (H + -CH20H), lettled, and this resulted in a yellow spot with vanillin / 244 (M + -CÛ2CH3), 219.

Spraying phosphoric acid and heating.

Example 8

The product was dissolved in 10 ml of dry tetrahydrofuran. Preparation of the benzhydryl ester of (aS, 5S) -chlorine-4,5-

added and treated with ethereal diazo-35 dihydro-a-phthalimide-5-isoxazole-acetic acid

(+) HN

May lex

To 192 u.1 (2 mmol) carbon tetrachloride in 25 ml of ethyl trotate and washing with 50 ml of tetrahydrofuran in a nitrogen atmosphere under Zim- 0.1N hydrochloric acid and then with a seed temperature 310 p.1 (1.9 mmol ) Hexamethyl-phosphorous sodium chloride solution and dried over sodium sulfate, phortriamide dichloride dropwise during 1 minute. After concentration in vacuo, the residue is added to 50 g of sili. 590 mg (1.3 mmol) of cagel are rapidly chromatographed within 2 minutes by eluting with (aS, 5S) -Oxo-cc-phthalimide-5-isoxazo-ethyl ester / hexane (200 ml elution) with 30% acetic acid benzhydryl ester , which gives lidin-acetic acid in about 5 ml of dry tetrahydrofuran 200 mg of crude product (33% yield) in addition to 245 mg. The heterogeneous solution is at 45 ° C during 48 starting material (42%). The crude product gave (cc5.5S) stirring for -3 hours. Alternatively, 590 mg (1.3 60 chloro-4,5-dihydro- <x-phthalimide-5-isoxazole-acetic acid in mmol) (aS, 5S) -Oxo-a-phthalimide-5-isoxazolidine in 20 ml form of needles with a melting point of 178-179 ° C dry tetrahydrofuran under a nitrogen atmosphere during recrystallization from methanol.

Room temperature 610 mg (2.6 mmol) of hexamethylphosphor- The thin layer chromatography gave an Rf value for triamide dichloride [made from hexachloroethane in the product of 0.70 in 35% ethyl acetate / hexane acetonitrile with hexamethylphosphorus triamide corresponding to 65 and an Rf value of 0 , 11 for the source material, the work of R. Appel and H. Schöler, Chem. Ber., 110, The optical rotation was:

2382 (1977)]. The solution is returned to

river heated. The reaction mixture is dissolved in 150 ml of vinegar- (01) 20 (CHCb) + 79 ° (589 nm), 82 ° (578), 94 ° (547), 163 ° (436),

43

646 961

256 ° (365)

NMR (CDCb): 7.80 (4H, m, PHTH), 7.32 and 7.22 (ÎOH, s, 0), 6.95 (1 H, s, 0CH), 5.6 (1H, m , CHO), 5.26 (1H, d, aH), 3.32 (2H, d, CH2).

Elemental analysis showed for C26H19CIN2O5

Calc .: C 65.75; H 4.03; N 5.90 Found: C 65.45; H 4.08; N 5.95.

(+)

CO2 CH ^ s

Using the procedures described above, but using the benzhydryl ester of (aS, 5S) -3-oxo-4,5-dihydro-4ß-phthalimid-5a-isoxazole-acetic acid of the formula VIIb, the benzhydryl- s ester of (aS, 5S) -3-chloro-4,5-dihydro-4ß-phthalimid-5a-iso-xazole-acetic acid (formula VIIIc, X = Cl).

Example 9

Preparation of (aS, 5S) -chloro-4,5-dihydro-a-phthalimide-10 5-isoxazole-acetic acid

COpH

In 185 mg (0.4 mmol) of benzhydryl ester of 3-chloro-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid in 3 ml of dry nitromethane in an ice bath under a nitrogen atmosphere, dry hydrogen chloride gas is introduced for 5 minutes . The bath is removed and the solution is stirred for 1 hour. After concentrating the solution in vacuo, the residue is chromatographed on CC-4 silica gel (30 g) by eluting with 30% ethyl acetate / hexane to give 115 mg of a white foam. This foam is recrystallized from hexane / ethanol in a ratio of 4: 1, whereby the (aS, 5S) -3-chloro-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid with a melting point of 176-177 ° C, which forms a hydrate when exposed to moisture.

The analytical results were as follows:

TLC: Rf = 0.45 (AIX).

NMR (acetone-dô): 7.95 (4H, s, PHTH), 5.6 (IH, m, CHO),

C02CH ^ a

25 5.27 (IH, d, a-H), 3.57 (2H, d, CH2).

The elementary analysis for C13H9CIN2O5 showed:

Calc .: C 50.58; H 2.94; N 9.08 30 Found: C 50.57; H 3.06; N 9.46.

Using the same procedure as described above, but using the benzhydryl ester of (aS, 5S) -3-chloro-4,5-dihydro-4ß-phthalimid-5a-iso-35 xazole-acetic acid of formula VIIIc (X means chlorine ) gives (aS, 5S) -3-chloro-4,5-dihydro-4ß-phthalimid-5a-iso-xazole-acetic acid (formula VIlb, X means chlorine).

1 Example 10

Preparation of (aS, 5S) -bromo-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid.

Br

In 185 mg (0.4 mmol) of benzhydryl ester of (aS, 5S) -chloro-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid in 3 ml of dry nitromethane in an ice bath, under a nitrogen atmosphere for 5 Minutes of dry hydrogen bromide gas introduced. The bath is removed and the solution is stirred for 1 hour. After concentrating the solution in vacuo, the residue is chromatographed on CC-4 silica gel (30 g) with 30% ethyl acetate / hexane to give 115 mg of a white foam. This foam is recrystallized from hexane / ethanol in a ratio of 4: 1, whereby the 3-bromo-4,5-dihydro-a-phtha-limid-5-isoxazole-acetic acid is obtained.

The elementary analysis showed for CnHîBr ^ Os:

Calculated: C 44.21; H 2.57; N 7.93 "Found: C 44.49; H 2.64; N 7.83.

The optical rotation was:

[a]: o (MeOH) + 67 ° (589 nm), 70 ° (578), 79 ° (547), 131 ° (436), 65 187 ° (365).

Using the same procedures as in Example 10, but using hydrogen iodide

646 961

44

3-iodo-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid was obtained instead of broxnhydrogen.

Likewise, using the procedure of

Example 10, but using hydrogen fluoride instead of hydrogen bromide to obtain 3-fluoro-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid.

Example 11

Methyl ester of phthalyl- (aS, 5S) -a-amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid from methyl phthalyl tricholomate

To 60 mg (0.2 mmol) of tricholomic acid ester phthalimide in 1 ml of dry tetrahydrofuran at room temperature under a nitrogen atmosphere are added 25 μl of carbon tetrachloride and then 30 μl of hexamethyl-phosphortriamide dichloride. After 16 hours, a further 10 μl-1 carbon tetrachloride and 10 μl of phosphine are added. The solution was worked up with ethyl acetate / water after a total reaction time of 36 hours. The products are chromatographed on 5 g of silica gel (50% E / H), whereby 35 mg of the methyl ester of phthalyl- (a5,5S) -a-amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid are obtained.

The analytical data were the following:

NMR: (Deuterochloroform) identical to AT-125-PHTH-2s methyl ester, made from natural (+) AT-125 according to Example 7.

TLC: Identical to AT-125-PHTH methyl ester (UV detection and staining identical).

30th

Example 12

(aS, 5S) -a-Amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid or AT-125

(+)

o2h

Ibi

(+ -)

+ nh-

I a4

260 ml of hydrazine hydrate are added to 710 mg (2.3 mmol) of 3-chloro-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid in 15 ml of water and the solution is stirred at 50 ° C. for 7 hours touched. After cooling, the solution is adjusted to pH 5.5 with acetic acid (80jj.1) and filtered and the precipitate is washed with 13 ml of water. The filtrate is diluted with 200 ml of 2-butanol and allowed to crystallize in the refrigerator for 18 hours. The filtration gives 240 mg (ccS, 5S) -a-amino-3-chloro-4,5-dihydro-5-iso-xazole-acetic acid and the concentration of the mother liquors by 15% gave 50 mg of a second yield of (aS, 5S) -a-amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid (71%).

The analytical values were as follows:

TLC: Rf = 0.40 (60/20/15/5; MEK / Acet / I-hO / AcOH). NMR: (DzO) 5.2 (1H, m CHO), 4.17 (1H, d, a-H), 3.61 (2H, d, CHa).

55

60

65

Using the same procedure as described above, but using DL- or D-3-chloro-4,5-dihydro-4ß-phthalimide-5a-isoxazole-acetic acid (formula VIlb, X means chlorine) gives DL in each case -or D-3-chloro-4,5-dihydro-4ß-amino-5a-isoxazole-acetic acid (formula Villa). A similar substitution in the compound of formula IIa to the compound of formula Ib gives tricholomic acid.

The analytical values were as follows;

Elemental analysis calculated for C5H9CIN2O3:

Calc .: C 33.63; H 3.95; N 15.69; Cl 19.86 Found: C 33.60; H 4.11; N 16.15; Cl 19.63.

CD: 02i6X = 13,300.

45 646961

Example 13

(aS, 5S) -a-Amino-3-bromo-4,5-dihydro-5-isoxazole-acetic acid

260 μl of hydrazine hydrate are added to 710 mg (2.3 mmol) of 3-bromo-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid in 15 ml of water and the solution is stirred at 50 ° C. for 7 hours touched. After cooling, the solution is adjusted to pH 5.5 by adding acetic acid (80 p.1) and filtered and the precipitate is washed with 13 ml of water. The filtrate is diluted with 200 ml of 2-butanol and allowed to crystallize in the refrigerator for 18 hours. Filtration gives 240 mg (aS, 5S) -a-amino-3-bromo-4,5-dihydro-5-isoxazole-acetic acid and concentration of the mother liquors by about 15% gave a further 50 mg of a second fraction (aS, 5S ) -A-Amino-3-bromo-4,5-dihydro-5-iso-xazole-acetic acid (71%).

The thin-layer chromatographic and nuclear magnetic resonance spectra results were indistinguishable from the chlorine analogues. The UV spectrum showed the same

Extinction but at 214 nanometers.

CD: 0213 "= 11 250.

Elemental analysis gave the following values for 25 CsH7BrN202:

Calculated: C 26.92; H 3.16; N 12.56; Br 15.73 Found: C 26.63; H 3.33; N 12.60; Br 15.58.

30 (The value for bromine was determined based on the culometric chlorine determination.)

Mass spectrum: The disilyl compound showed a peak M + -15 at 352 and 543 m / e.

35 Example 14

Direct conversion of tricholic acid to AT-125

+ NH3

+ NH;

! fd

! a 4,

To 140 mg of tricholomic acid in 3 ml of POCb under a nitrogen atmosphere at room temperature was added 160 ml of purified diethylaniline. The mixture was placed in a water bath at 100 ° C for 5 minutes. The POCb was removed in vacuo. The solid so obtained was treated with 2 ml of water and stirred for 10 minutes while neutralizing with concentrated ammonium hydroxide solution. After the material had been lyophilized, it was applied to a column packed with 10 g of silica gel and eluted with 85% ethyl acetate / 5%

Water / 10% acetic acid. A dark band was found, which had a thin layer chromatographic mobility, similar to AT-125. The material was examined for its biological activity against B. subtilis (syn.) And it was shown that it had at least a 2% activity, assuming that the tricholomic acid used as the starting material was 100% pure and the biological Tricholomic acid activity.

60

646 961 46

Example 15

Benzhydryl ester of (aS, 5S) -methoxy-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid

Ha le (X = OCHa)

To 90 mg (0.2 mmol) of benzhydryl ester of 3-oxo-a-phthalic. The analytical data were the following: imide-5-isoxazolidine-acetic acid (formula IIa) in 5 ml of methylene chloride, a 50% molar excess of more common NMR (CdCb): 7.80 (m, 4H), 7.32 and 7.22 (s, 10H, Ph), 6.96 ethereal diazomethane solution added. A lot of (s, Ph: CH), 5.1 -5.8 (m, 2H), 3.75 (s, CH3O), 3.07 (d, J = 7.5 5 µl ßFs.EtzO added and after 30 minutes the 25 Hz, 2H).

The reaction mixture is concentrated and the residue is brought to TLC (AIX) = Rf = 0.60.

a preparative thin layer chromatography plate (AIX eluent), whereby one

45 mg benzhydryl ester of 3-methoxy-4,5-dihydro-a- A second product is obtained in an amount of 35 mg phthalimide-5-isoxazole-acetic acid. 30 formed, which has the structure and has the following analytical data: 1H), 5.1-5.9 (m, 2H), 3.10 (s, NCH3), 2.81 (m, 2H).

NMR (CDCb): 7.82 (m, 4H), 7.32 and 7.22 (s, 10H), 6.97 (s, TLC (AIX): Rf = 0.40.

Example 16

Methyl ester of phthalyl- (aS, 5S) -cc-amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid

To 110 mg (0.35 mmol) of phthalimide in 3 ml of ether was added NMR (CDCb): 7.87 (4H, m 0), 5.6 (IH, m, CHO), 5.22 (IH, ethereal diazomethane) until gas evolution d, J = 6 Hz, aH), 3.78 (3H, s, CH3), 3.40 (2H, d, J = 9, CH2).

has stopped and a yellow color persists. Concentrate 6s TLC: (AIX) Rf = 0.50 (85% E / H) Rf = 0.70. and chromatography on 15 g of silica gel (40% acetic acid-13C NMR: (CDCb) 167.5 (CO 2), 167.1 (CON), 149.9 (C-Cl), ethyl ester / hexane elution) gave 110 mg of an oil (95%). 134.7, 131.5, 124.0 (0), 80.2 (CHO), 53.1 (CH3), 52.9 (a-C). The analytical data were as follows: 41.5 (CH2).

47

646 961

Analysis:

Calculated: C 52.10; H 3.44; N 8.68 Found: C 52.58; H 3.67; N 8.39.

Mass spectrometry: no M + m / e 322.291 (M + -OCH3), 287 (M + -C1), 263 (IVP-CH2CH3), 219 (62% PHTH-COCHO2CH3).

Example 17

3-Chloro-a - {[(9H-fluoren-9-yl-methoxy) carbonyl] amino} -4,5-dihydro-5-isoxazole-acetic acid.

300 mg of sodium bicarbonate were added to 178 mg of AT-125 in 4 ml of water. After the mixture had cooled to 0-5 ° C., 300 mg of 9-hydroxymethylfluorenylcarbo-

nyl chloride was added and the mixture was stirred for 2 hours and left to stand at room temperature overnight. The semi-solid thus obtained is taken up in ethyl acetate and 1N hydrochloric acid 5N is added until a pH of 3 is reached. The organic phase is separated, dried over sodium sulfate, concentrated and chromatographed on 20 g of CC-4 silica gel, using 40% ethyl acetate / hexane as the eluent. Those fractions containing the desired product are evaporated and the residue is recrystallized from ethyl acetate, whereby 3-chloro-a - {[(9H-fluoren-9-yl-methoxy) carbonyl] amino} -4,5- dihydro-5-isoxazole-acetic acid with a melting point of 182-183 ° C. The nuclear magnetic resonance spectrum ls corresponds to the desired structure.

B

Claims (14)

646 961 2nd PATENT CLAIMS 1. Derivatives of isoxazoline-5-acetic acid, characterized in that they are optically active isomers of a compound of formula I COaR (I) , R i or racemic mixtures of the optically active isomers of the formula I or salts of these compounds, wherein in formula I R represents a hydrogen atom, an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms and with 1-5 carbon atoms or an aralkyl group with 7-20 carbon atoms, X is a bromine atom, a chlorine atom or a grouping of the formulas -ORi, -SRi, or -NR'R " is, in these groups Ri represents an alkyl group with 1-12 carbon atoms, an aryl group with 6-20 carbon atoms or an aralkyl group with 7-20 carbon atoms, R 'and R "are identical to or different from one another and denote hydrogen atoms or alkyl groups with 1-8 carbon atoms, Ru and Ris independently represent hydrogen atoms or groups of the formula in which Rs has the following meanings: (a) a grouping of the formula 5 R9 R11 I I -C-C- I I 10 Rio R.12 in which R9, Rio, Rh and R12 independently of one another have the meaning 15 of hydrogen atoms or alkyl groups having 1-5 carbon atoms or (b) a grouping of the formula 20 V / CH2 \ CH CH0 i i 2 * / CH2 2S C < or (c) orthointerphenylene or (d) means a substituted orthointerphenylene, under 30 provided that if all R, Ri4 and Ris Hydrogen atoms mean that X must not be a chlorine atom and provided that if X is a chlorine atom and R represents either a hydrogen atom or a lower alkyl group with 1-8 carbon atoms, 35 neither Ri4 nor Ris may be an alkoxycarbonyl group, nor R14 and R15 together with the nitrogen atom to which they are attached is a grouping of the formula O O 40 Are -COR6, -C-Rt, wherein Ró is an alkyl group with 1 -8 carbon atoms, a halo-45 alkylated group with 1-3 halogen atoms and 1-5 carbon atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7- Represents 20 carbon atoms and R7 represents an alkyl group with 1-12 carbon atoms, an aryl group with 6-20 carbon atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7-20 carbon atoms, or R14 and R15 together with the nitrogen atom to which they are attached , a grouping of the formula ss -N are allowed to form in which Rs has the meaning given above, or that they are optically active isomers of a compound of the formula VIII 0 R 13 ■ ù 4r15 Vlld 60 ■ COzR or racemic mixtures of the optically active isomers of the formula VIII or salts of these compounds, where 6s in the formula VIII R "" is an alkyl group with 1-1-2 carbon atoms, a halogenated alkyl group with 1-5 carbon atoms and 1-3 halogen atoms, an aralkyl group with 7-20 carbon atoms. 3rd 646961 Means atoms or a substituted aralkyl group with 7-20 carbon atoms, Ru 'and Ris' are hydrogen atoms or groups of the formula O O II II -CORó, -C-Ri ' mean with the proviso that at least one of the substituents Rh 'and Ris' is hydrogen, and in which Rô has the same meaning as in formula I and R7' is an alkyl group with 1-8 carbon atoms, an aryl group with 6- 20 carbon atoms, an aralkyl group having 7-20 carbon atoms or a substituted aralkyl group having 7-20 carbon atoms, or R14 'and Ris' together with the nitrogen atom to which they are attached is a grouping of the formula 0 0 form what Rs has the same meaning as in formula I and R13 is a hydrogen atom, an alkoxycarbonyl group, a halogenated alkoxycarbonyl group or an aralkoxycarbonyl group, provided that when R13, Rm 'and Ris all represent hydrogen, R "" does not Is alkyl group of 1-8 carbon atoms, and when Rb is a halogenated alkoxycarbonyl group or an aralkoxycarbonyl group, neither the group of the formula O II -CORs can be an aralkoxycarbonyl group or a halogenated aralkoxy-carbonyl group, nor the group of the formula O II -C-RT ' may be an aralkylcarbonyl group or a halogenated alkylcarbonyl group. 2. derivatives of isoxazoline-5-acetic acid according to claim 1, characterized in that they are optically active isomers of the formula I or racemic mixtures of the optically active isomers or salts of these compounds, in the formula I X represents a bromine atom or chlorine atom. 3. derivatives of isoxazoline-5-acetic acid according to claim 2, characterized in that in them Rw and Ris are hydrogen atoms or Rm and Ris together with the nitrogen atom to which they are attached, a grouping of the formula 0 0 represent, wherein Rs has the meanings given in claim 1, provided that, however, when Ru and Ru together with the nitrogen atom form this cyclic grouping and X is also a chlorine atom, the radical R must have a different meaning than that of a hydrogen atom or an alkyl group with 1-8 carbon atoms. 4. Connection according to claim 3, characterized in that it: the (aS, 5S) -a-amino-3-bromo-4,5-dihydro-5-isoxazole-acetic acid of the p-methoxybenzyl ester of (aS, 5S) -amino-3-chloro-4,5 - dihydro-5-isoxazole acetic acid, the p-methoxybenzyl ester of N-phthaloyl- (aS, 5S) -a- amino-3-chloro-4,5-dihydro-5-isoxazole-acetic acid, the benzhydryl ester of (ctS, 5S) -3-chloro-4,5-dihydro-a- phthalimide-5-isoxazole-acetic acid or the (aS, 5S) -3-bromo-4,5-dihydro-a-phthalimide-5-isoxazole- is acetic acid. 5. derivatives of isoxazoline-5-acetic acid according to claim 1, characterized in that they are optically active isomers of the compound of formula I or racemic mixtures of the optically active isomers of formula I or salts of these compounds, wherein in formula I. X is a grouping of the formula -ORi is where Ri has the meanings given in claim 1. 6. A compound according to claim 5, characterized in that Rwund Ris together with the nitrogen atom to which they are attached, a grouping of the formula 0 0 form, wherein Rs has the same meaning as in claim 1. 7. A compound according to claim 6, characterized in that in it R is a benzhydryl group, R14 and R15 together with the nitrogen atom to which they are attached form a grouping of the formula 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 646961 4th 0 I " or racemic mixtures of the optically active isomers of the compounds of the formula I "or salts of these compounds, in which Rs form an orthointerphenylene group of the structure R 'an alkyl group having 1-12 carbon atoms, an aryl 15 group with 6-20 carbon atoms or an aralkyl CH. means group with 7-20 carbon atoms, thereby \. /% s. characterized in that a compound of formula II CH CH 1H CH ^ v / 20th and X is a methoxy group, so that this compound is the benzhydryl ester of (aS, 5S) -3-methoxy-4,5-dihydro-a-phthalimide-5-isoxazole-acetic acid. 8. Process for the preparation of optically active isomers of the formula I ' 25th II 30th wherein R '"represents an alkyl group of 1-8 carbon atoms, a halogenated alkyl group of 1-3 halogen atoms and 1-5 carbon atoms or an aralkyl group of 7-20 carbon atoms, chlorination to form a compound of formula I' or racemic mixtures of the optically active isomers of the compounds of the formula I ', in which R' "denotes an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms and with 1-5 carbon atoms or an aralkyl group with 7-20 carbon atoms, characterized in that a compound of formula II H where R '"has the same meaning as in formula II, subjects these esters of formula I' to form the corresponding free acid of the formula II wherein R '"has the same meaning as in formula I' subject to chlorination. 9. Process for the preparation of optically active isomers of the formula I " 65 saponified, and this with an alkali metal salt of a mercapane of the formula Ri-S-M where Ri has the same meaning as in formula I " and the protective group on the amino group is split off. 10. Process for the preparation of optically active isomers of the formula I '" nowhere 'nh; or racemic mixtures of the optically active isomers of the compounds of the formula I '"or salts of these compounds, in which R 'and R "are identical to or different from one another and denote hydrogen atoms or alkyl groups with 1-8 carbon atoms, characterized in that a compound of the formula II wherein R '"represents an alkyl group of 1-8 carbon atoms, a halogenated alkyl group of 1-3 halogen atoms and 1 -5 carbon atoms or an aralkyl group of 7-20 carbon atoms, chlorination to form a compound of formula I' Cl in which R '"has the same meaning as in formula II, then subjects this ester to form the corresponding free acid of the formula Cl 646961 saponified and this with ammonia or an amine of the formula H-N <T \ R " wherein R 'and R "have the same meaning as in formula Ic, and the amino protecting group is split off. 11. Process for the preparation of an optically active isomer of the formula I "" 0-ch or racemic mixtures of the optically active isomer of the formula I "" or salts, characterized in that a compound of the formula II HN 0 wherein R '"is an alkyl group with 1-8 carbon atoms, is a halogenated alkyl group with 1-3 halogen atoms and 1-5 carbon atoms or an aralkyl group with 7-20 carbon atoms, reacted with diazomethane in a solvent with a catalytic amount of boron trifluoride diethylate and cleaves the amino protecting group and the group R '". 12. Process for the preparation of optically active isomers of the formula I ' Cl or racemic mixtures of the optically active isomers of the compounds of the formula I ', in which R' "denotes an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms and with 1-5 carbon atoms or an aralkyl group with 7-20 carbon atoms and optically active isomers of the formula VH'd 5 s 10th 15 20th 25th 30th 35 40 45 50 55 60 65 646 961 6 «Jy / -CH2OC-NS J 0— ( is obtained, the compound of the formula VH'd is isolated from this mixture and the benzyloxycarbonyl protective group is split off from the remaining compound of the formula Ile and thus a compound of the formula IIb VI \ d C02H or racemic mixtures of the optically active isomers of the compounds of the formula VH'd, characterized in that a compound of the formula lila io \ 0 15 practice 20th oH JL ~ <^ ch2oÛno ^ nÇJO) 0 receives this to the ester of formula IIa purple 25 H 30th with a ruthenium compound in the presence of an oxidizing agent and thereby a mixture of the compound of the formula Ile ! la «JP 0 wherein R '"has the same meaning as in formula I', esterified and subjecting this ester to chlorination to form the compound of formula I '. 13. A composition for combating the growth of microorganisms, characterized in that it contains, as an active ingredient, a derivative of isoxazoline-5-acetic acid, which is an optically active isomer of a compound of the I IC «Formell or Vlld with the compound of the formula Vll'd C02R nri4r15 (X) 60 0 VI fd \ no; 4r15 (Vlld) C02H • C0aR ' 7 646 961 or racemic mixtures of the optically active isomers of the formula I or VIII or a salt of the compounds of the formula I, where in the formulas IR is a hydrogen atom, an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms and also with 1-5 carbon atoms or an aralkyl group with 7-20 carbon atoms, Rh and Ris independently of one another are hydrogen atoms or groups of the formula O O II II -CORó, -C-R7 mean, wherein Ró represents an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms and 1-5 carbon atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7-20 carbon atoms and R7 denotes an alkyl group with 1-12 carbon atoms, an aryl group with 6-20 carbon atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7-20 carbon atoms or Ri4 and R15 together with the nitrogen atom to which they are attached, a grouping of the formula 0 0 represent what Rs has the following meanings: (a) a grouping of the formula R9 Ru 1 I -C-C- I I Rio R12 in which R9, Rio, Ri 1 and R12 independently of one another have the meaning of hydrogen atoms or alkyl groups having 1-5 carbon atoms or (b) a grouping of the formula ch ch0 I I 2 ™ ch2 nch ( or (c) orthointerphenylene or (d) represents a substituted orthointerphenylene, and X represents a bromine atom, a chlorine atom or a grouping of the formulas -ORi, -SRi, or-NR'R " is, in these groups Ri represents an alkyl group with 1-12 carbon atoms, an aryl group with 6-20 carbon atoms or an aralkyl group with 7-20 carbon atoms, R 'and R "are identical to or different from one another and denote hydrogen atoms or alkyl groups with 1-8 carbon atoms, provided that when all the radicals R, Ru and R15 denote hydrogen atoms, X must not be a chlorine atom and under the further condition that when X is a chlorine atom and R represents either a hydrogen atom or a lower alkyl group with 1-8 carbon atoms, neither R14 nor Ris may be an alkoxycarbonyl group, nor R14 and R15 together with the nitrogen atom to which they are attached are a grouping of the formula 0 n s 0 may form, in which Rs has the meaning given above and in formula Vlld R "" denotes an alkyl group with 1-12 carbon atoms, a halogenated alkyl group with 1-5 carbon atoms and 1-3 halogen atoms, an aralkyl group with 7-20 carbon atoms or a substituted aralkyl group with 7-20 carbon atoms, R14 'and Ru' are hydrogen atoms or groups of the formula O O II II -COR6, -C-Rt ' with the proviso that at least one of the substituents R14 'and R15' is hydrogen, and in which Rô has the same meaning as in formula I and R7 'is an alkyl group with 1-8 carbon atoms, an aryl group with 6 Is -20 carbon atoms, an aralkyl group having 7-20 carbon atoms or a substituted aralkyl group having 7-20 carbon atoms, or R14 'and R15' together with the nitrogen atom to which they are attached is a grouping of the formula 0 n s 0 5 10th 15 20th 25th 30th 35 40 45 50 55 60 65 646 961 8th form what Rs has the same meaning as in formula I and Ri3 is a hydrogen atom, an alkoxycarbonyl group, a halogenated alkoxycarbonyl group or an aralkoxycarbonyl group, provided that when R13, Ru 'and Ris' are all hydrogen, R "" does not Is alkyl group of 1-8 carbon atoms, and when R13 is a halogenated alkoxycarbonyl group or an aralkoxycarbonyl group, neither the group of the formula O II -COR « can be an aralkoxycarbonyl group or a halogenated aralkoxycarbonyl group, nor the group of the formula O II -C-R7 ' may be an aralkylcarbonyl group or a halogenated alkylcarbonyl group. (VIId) VU ' co2r NR14R1 (I) ■ COaR or racemic mixtures of the optically active isomers 10 of the compounds of the formula I or VIII or salts of these compounds. In the formula I, R here denotes a hydrogen atom, an alkyl group with 1-8 carbon atoms, a halogenated alkyl group with 1-3 halogen atoms, and also with 1-5 carbon atoms or an aralkyl-15 group with 7-20 carbon atoms and R14 and Ris independently of one another represent hydrogen atoms or groups of the formula O O 20th