DE3220672A1 - Cis-oxazaphosphorine-4-thioalkanesulphonic acids, their neutral salts, process for the preparation thereof and pharmaceutical preparations containing them - Google Patents

Abstract

The present invention relates to the novel cis-compounds of oxazaphosphorine-4-thioalkanesulphonic acids and neutral salts thereof of the general formula I <IMAGE> processes for their preparation and pharmaceutical preparations containing them.

Description

cis-Oxazaphosphorin-4-thio-alkanesulfonic acids, their

neutral salts, processes for their preparation and containing them pharmaceutical preparations.

Addition to DE-PS ........ (Patent application P 31 11 428.8 of March 24, 1981) The present invention relates to the new cis-compounds of oxazaphosphorin-4-thio-alkanesulfonic acids and their neutral salts of the general formula I. where R1, R2 and R3, which can be identical or different from one another, represent hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl and at least two of these radicals are 2-chloroethyl and / or 2-methanesulfonyloxyethyl, R4 is hydrogen or methyl is, X is a straight or branched-chain C26-alkylene, and the hydrogen cation, an alkali or alkaline earth cation, the guanidinium, morpholinium or cyclohexylammonium cation or the cation which is derived from an amine of the formula NR5R6R7, in which the radicals R5 to R7 are the same or different and are hydrogen, C1-C2-alkyl groups or oxyethyl groups, or where Y + is the ethylenediammonium cation H3N + -CH2CH2-N + H3 or the piperazonium ion and z is 1 if is a monobasic cation, or 2 is if a dibasic cation or the cation of a compound having two monobasic cations.

Because of their good accessibility and good properties are included those compounds of formula I preferred in which Y is the guanidinium, morpholinium or cyclohexylammonium cation or the cation which is derived from an amine Formula NR5R6R7 derives, in which the chain R5 to R7 are identical or different and hydrogen, C1-C-alkyl groups or oxyethyl groups or in which Y denotes Ethylenediammonium cation H3N + -CH2CH2-N + H3 or the piperazonium cation and z is 1 is when Y is a monobasic cation, or is 2 when YO is a dibasic cation c'dcr is the cation of a compound with two monobasic cations.

If Y + is a cation which is different from an amine of the formula NR5R6R7 derives, it is preferably the ammonium cation or one with 1-3 C1-C2-alkyl groups substituted ammonium cation or one with 1-3 2-hydroxyethyl groups substituted ammonium cation.

Potassium and alkaline earth salts also have other beneficial properties of the acids of the formula I and are therefore preferred.

Among the acids of the formula I neutralized with nitrogen bases the ammonium salts in which Y + is the N114 cation are particularly preferred, the cyclohexylammonium salts, in which Y + is the C6H11NH3 + cation, or the guanidinium salts, in which Y + is the N + H2 = (CH2) cation.

The salts according to the invention are particularly easily accessible and therefore preferably cis-sodium 2- [2- (bis- (2-chloroethyl) -amino) -2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorine 4-yl] thioethanesulfonate and the corresponding ammonium, cyclohexylammonium and Potassium salt.

Of particular importance are those compounds of the formula I in which R1 and R2 are the 2-chloroethyl group, R3 and RÄ are hydrogen, X is the ethylene group and z = 1 and Y + = NH4 +, or N + H2 = (CH2) 2 or x = (CH2) 3, z = 1, where Y = N + H2 = C (NH2) 2.

If Y is an alkaline cation, it is in particular that Sodium or potassium salt; if Y is an alkaline earth, it is in particular around the neutral calcium or magnesium salt; if Y + is a cyclohexylammonium cation is the following cation: C6H11NH3; if Y is the cation R5R6R7N + this is derived in particular from the following amines: methylamine, ethylamine, Dimethylamine, diethylamine, trimethylamine, triethylamine, methylethylamine, dimethylethylamine, Diethylmethylamine, 2-hydroxy-ethylamine, bis- (2-hydroxy-ethyl) -amine, tris- (2-hydroxy-ethyl) -amine, (2-hydroxy-ethyl) -methylamine, (2-hydroxy-ethyl) -dimethylamine, bis- (2-hydroxy-ethyl) -methylamine, (2-hydroxy-ethyl) -ethylamine, (2-hydroxy-ethyl) -diethylamine, bis- (2-hydroxy-ethyl) -ethylamine, (2-hydroxy-ethyl) -methylethylamine.

In general, the synthesis according to DE-PS .... (patent application P 31 11 428.8, in which a 4-hydroxy- or 4-C14-alkoxy-oxazaphosphorine of the general formula II in which R1, R2, R3 and R4 have the same meaning as in formula I and Z is hydrogen or C14 -alkyl, with a compound of the general formula III HS-X-SO2-Y + III in which X and Y + have the same meaning as in formula I have, and, if Y + is hydrogen, and the oxazaphosphorin-4-thio-alkanesulfonic acid obtained are neutralized with a base corresponding to the other meaning of Y +, cis / trans mixtures. In the case of compounds that crystallize well, this gives the cis-form crystallized. However, if the reaction is carried out in anhydrous solvents or in solvents with a low water content, the cis form is obtained exclusively or predominantly. For example, the pure cis form of a compound of formula I that does not crystallize or that crystallizes poorly can be prepared by adding an acetone solution of the compound of formula II to an aqueous solution of the compound of formula III at temperatures between -30 and + 200C and after is dropped several times after completion of the implementation.

The starting compounds according to formula II can be used as racemic cis isomers (See above for illustration) as an optically active cis-shape of which used will. Optically active compounds according to formula II are obtained, for example, from the salts of the optically active sulfonic acids (see above for representation) by adding to the corresponding optically active 4-hydroxy - oxazaphosphorines of the formula II hydrolyzed (in water for example between 0 and 50 ° C, especially 5 to 30 ° C) and the released thiol of the formula III is oxidized to disulfides (for example using iodine or hydrogen peroxide in water). Another procedure goes for example from a separate optically active cyclophosphamide derivative Example NrO 16 or 33 of the German patent application P 31 33 309.5 and hydrolyzed this compound to optically active 4-hydroxy-cyclophosphamide. In another Process is, for example, optically active Cyclophosphiiamid (DE-OS 29 44 106) in a method described (Peter et al, Cancer Treatment Reports 60, 429 (1976)) converted to optically active 4-Ilydroxycyclophosphamid.

For example, optically active bases can be used for the resolution of the racemate for example 1-phenylethylamine, brucine, quinidine, strychnine and cinchonine as well other bases and methods in question are discussed in "Optical Resolution Procecures for Chemicals Compounds, "Vol. 2, Paul Newman, 1981 Publishers Optical Resolution Information Center in Rivexdale, USA. These optically active bases can also be used for the synthesis according to the reaction of a compound of the formula II with a compound of the formula III can be used instead of the meanings of Y already given. In this case, this optically active base is then opposed in the usual way Hydrogen or another base Y according to the definition already given by Y to exchange.

The compounds according to the invention can be used in the fight against Cancer and immunosuppression. They have opposite the cis / trans mixtures or the trans compounds have increased potent anti-tumor activity. they draw by a high activity with parenteral and with oral application as well characterized by low general toxic phenomena.

They have a high carcinotoxic selectivity in vivo and in in vitro a high cytotoxic specificity.

The substances were used to determine the carcinotoxic effects tested in animal experiments on a number of tumors of different chemoresistance. For the evaluation, the relationship between the Logarithms of the doses and the frequencies of cured and surviving test animals the mean curatively effective doses (DC 50 tig / kq /)) were determined. As a reference substance, which is close to the products according to the invention in chemical constitution, the commercially available compound cyclophosphamide was used.

In the case of lymphatic leukemia L5222 of the rat (strain: BD IX) was this mean curative dose - with a single intravenous application on the 5th Day after inoculation of the leukemia - for the products according to the invention as well as for cyclophosphamide at 1.5 mg / kg.

In the chemosensitive Yoshida ascites carcinosarcoma (line ER 13) of the Rat (strain: Sprague-Dawley) this DC 50 was for the products according to the invention as well as for the comparison substance cyclophosphamide at 1 mg / kg.

The toxic effect was tested in an analogous manner, whereby from the Relationship of the logarithms of the doses and the frequency of death the mean lethal Dose (DL 50 [mg / kg]) was determined.

For the products according to the invention, this DL 50 is once intravenous application around 300 mg / kg. The comparatively determined mean lethal Single intravenous dose of cyclophosphamide is 244 mg / kg. The products according to the invention accordingly have an acute acute level that is around 20% lower Toxicity or

with the same curative effectiveness a correspondingly higher therapeutic one Broad.

For the study of the cytotoxic specificity in vitro were each approx. 3. 10 'cells obtained aseptically from the chemoresistant Yoshida ascites carcinosarcoma (Line AH13, strain ASTA) with increasing concentrations of the products according to the invention incubated for two hours at 37 ° C and washed out several times into the abdominal cavity implanted in untreated host animals. From the relationship between the logarithms the concentrations and frequencies of developing tumors was using the probit analysis determined the mean cytotoxic doses (CE50 [µg / ml]). Under The CE50 of the products according to the invention lies between these test conditions 3 and 5 µg / ml.

Because cyclophosphamide is initially an inactive form of transport compound exhibits no cytotoxic activity, was used as a comparison substance in these studies the active one primary metabolite 4-hydroxy-cyclophosphamide used, which is formed in the body through enzymatic activation, mainly in the liver will. The mean cytotoxic concentration is for this comparison substance also at 5 jig / ml.

The compounds of the invention are pharmaceutical for the manufacture Compositions and preparations suitable. The pharmaceutical compositions or medicaments contain one or more of the substances according to the invention as the active ingredient Compounds, if necessary in a mixture with other pharmacological respectively pharmaceutically active substances. The pharmaceuticals are manufactured in a well-known manner Way, with the known and usual pharmaceutical substances as well as others Conventional carriers and diluents can be used.

Examples of such carriers and auxiliaries are those Substances in question that are listed in the following literature as auxiliaries for pharmacy, Cosmetics and related areas are recommended or indicated: Ullmanns Encyklopadie der technischen Chemie, Volume 4 (1953), pages 1 to 39; Journal of Pharmaceutical Sciences, Volume 52 (1963), page 918 and ff., H.v. Czetsch-Lindenwald, auxiliaries for pharmacy and related areas; Pharm. Ind., No. 2, 1961, page 72 and ff .; Dr. H. P. Fiedler, Lexicon of auxiliaries for pharmacy, cosmetics and related Territories Cantor KG. Aulendorf in Württemberg 1971.

Examples of this are gelatin, natural sugars such as cane sugar or milk sugar, lecithin, pectin, starch (e.g. corn starch), alginic acid, Tylose, talc, lycopodium, silica (for example colloidal), cellulose, cellulose derivatives (For example cellulose ethers, in which the cellulose hydroxyl groups are partially with lower saturated aliphatic alcohols and / or lower saturated aliphatic ethereal oxyalcohols, for example methylo>: propylcellulose) stearates, Magnesium and calcium salts of fatty acids with 12 to 22 carbon atoms, in particular the saturated (for example stearates), emulsifiers, oils and fats, in particular vegetable (for example peanut oil, castor oil, olive oil, sesame oil, cottonseed oil, Corn oil, wheat germ oil, sunflower seed oil, cod liver oil, mono-, di- and triglycerides of saturated fatty acids C12H24O2 to C18H36O2 and their mixtures), pharmaceutical compatible monohydric or polyhydric alcohols and polyglycols such as polyethylene glycols and derivatives thereof, esters of aliphatic saturated or unsaturated fatty acids (2 to 22 carbon atoms, especially 10 to 18 carbon atoms) with monovalent aliphatic Alcohols (1 to 20 carbon atoms) or polyhydric alcohols such as glycols, glycerine, Diethylene glycol, pentaerythritol, sorbitol, mannitol and so on, which may be can also be etherified, benzyl benzoate, dioxolanes, glycerine formals, tetrahydrofurfuryl alcohol, Polyglycol ether with C1-C12 alcohols, dimethylacetamide, lactamides, lactates, ethyl carbonates, Silicones (especially medium viscosity dimethylpolysiloxanes) magnesium carbonate and similar.

For example, water or physiological solutions are used to produce solutions compatible organic solvents in question, such as ethanol, 1,2-propylene glycol, Polyglycols and their derivatives, dimethyl sulfoxide, fatty alcohols, triglycerides, partial esters of glycerine, paraffins and the like In making the Preparations can contain known and customary solubilizers or emulsifiers, be used. As solubilizers and emulsifiers, for example, come in Question: polyvinylpyrrolidone, sorbitan fatty acid esters such as sorbitan trioleate, lecithin, Acacia, tragacanth, polyoxyethylated sorbitan monooleate, polyoxyethylated fats, polyoxyethylated olotriglycerides, linolized olotriglycerides, polyethyleneoxyå rondensation products of fatty alcohols, alkylphenols or fatty acids or 1-methyl-3- (2-hydroxyethyl) -imidazolidone- (2). Polyoxyethylated here means that the substances in question are polyoxyethylene chains contain, whose degree of polymerization is generally between 2 to 40 and in particular is between 10 and 20.

Such polyoxyethylated substances can, for example, by reaction of compounds containing hydroxyl groups (e.g. mono- or diglycerides or unsaturated compounds such as those that contain oleic acid residues) can be obtained with ethylene oxide (for example 40 moles of ethylene oxide per mole of glyceride).

Examples of oleopotriglycerides are olive oil, peanut oil, castor oil, Sesame oil, cottonseed oil, corn oil (see also Dr. H.P. Fiedler "Lexicon of auxiliaries for pharmacy, cosmetics and related areas 1971, pages 191 to 195).

In addition, the addition of preservatives, stabilizers, Buffer substances, e.g. calcium hydrogen phosphate, colloidal aluminum hydroxide, Flavor corrections, antioxidants and complexing agents (for example ethylenediaminetetraacetic acid) and the like possible. If necessary, it is used to stabilize the active ingredient molecule with physiologically compatible acids or buffers to a pH range of approx. 3 to 7 to be set. In general, a neutral to weakly acidic one is possible (up to pH 5) pH value preferred.

As antioxidants, for example, sodium metabisulphite, ascorbic acid, Gallic acid, gallic acid alkyl ester, butylhydroxyanisole, nordihydroguajaretic acid, Tocopherols as well as tocopherols + synergists (substances the heavy metals through complex formation bind, for example lecithin, ascorbic acid, phosphoric acid) for use. Of the The addition of the synergists increases the antioxidant effect of the tocopherols considerably.

Sorbic acid and p-hydroxybenzoic acid esters, for example, can be used as preservatives (for example lower alkyl ester), benzoic acid, sodium benzoate, trichloroisobutyl alcohol, Phenol, cresol, benzethonium chloride and formalin derivatives are possible.

The pharmaceutical and galenic handling of the invention Connections are made according to the usual standard methods. For example, are substance (s) and auxiliary or carrier substances by stirring or homogenizing (for example by means of a conventional mixer) well mixed, with in general at temperatures between 20 and 80 ° C, preferably 20 to 50 ° C, in particular is worked at room temperature. Otherwise, reference is made to the following standard work referenced: Sucker, Fuchs, Speiser, Pharmaceutical Technology, Thieme-Verlag Stuttgart, 1978.

The application of the active ingredients or the drugs can on the skin or mucous membrane or inside the body, for example oral, enteral, pulmonary, rectal, nasal, vaginal, lingual, intravenous, intraarterial, intracardiac, intramuscular, intraperitoneal, intracutaneous, subcutaneous, intrapleural, intrathecal, intracavity.

In particular, the addition of other active pharmaceutical ingredients, especially of uroprotectors such as the sodium salt of 2-mercaptoethanesulfonic acid or the disodium salt of the corresponding disulfide of the formula HO3S-CH2CH2SSCH2-SO3H, but other systemic or regional detoxifying agents are also possible or cheap.

The compounds of the invention show in intravenous, intraperitoneal or oral application to various experimental tumors of the rat and the Mouse good cytostatic and curative activity.

Examples of such indications are: leukemia, lymphogranulomatosis, Lymphosarcoma, reticulosarcoma, plasmacyitoma, M. Waldenström; Tumors with spread Growth, especially ovarian, breast, bronchial carcinoma, neuroblastic horn, dead tumors, Kidney carcinoma, pancreatic carcinoma, seminoma, Ewing sarcoma; Postoperative addition treatment, especially in the case of chemosensitive tumors, which presumably affect the local boundaries already exceeded and have a poor prognosis despite radical surgery.

Furthermore, the compounds according to the invention come in contrast to the known agents "cyclophosphamide; and" ifosfamide ", especially for the following Applications in question: 1) for regional perfusion in the extremities and the large body cavities, 2) for an in vitro treatment of the bone marrow in the extracorporeal treatment of the bone marrow of leukemia patients; 3) for pretherapeutic sensitivity tests of tumors in vitro.

For example, the compounds according to the invention are used in rats 5 days after intraperitoneal implantation of 105 cells of the L5222 leukemia with different doses administered intravenously, intraperitoneally or orally and depending on has a curative effect on the dose.

As a cure, the relapse-free and metastasis-free survival of the tumor-bearing animals defined after 90 days.

From the frequency of divisions obtained with the different doses is determined as the mean curative dose (DC50) using probit analysis according to R. FISHER Dose calculated with which 50% of the tumor-bearing animals can be cured * For example the compounds according to the invention are also intraperitoneally one day after Implantation of 106 cells of Yoshida ascites sarcoma AH13 at different doses administered intravenously, intraperitoneally or orally and depending on the dose achieved a curative effect.

Here, too, the curative effect is free of recurrences and metastases The life of the tumor-bearing animals is defined over 90 days.

In a corresponding manner, using probit analysis according to R. FISHER calculated as the mean curative dose (DC50) that dose at which 50% of the tumor-bearing Animals can be cured.

The compounds according to the invention are also used, for example once or several times (4 x) on consecutive days after intraperitoneal Implantation of 106 cells of mouse leukemia L1210 with different doses intravenously, administered intraperitoneally or orally and achieved a cytostatic effect.

The cytostatic effectiveness is expressed as an extension of the median survival time of tumor animals and is recorded as a dose-dependent percentage extension the survival time compared to an untreated control group.

The mean curative dose is independent for the rat tumors of the dosage form in the range from 0.1 to 10 mg / kg. With the same doses an extension of the median survival time of 100 ei for mouse leukemia L1210 achieve (see N. Brock: Pharmacological principles of cancer chemotherapy In: A. GEORGII (Ed), Negotiations of the German Cancer Society Volume 1, pp 15 - 42, Gustav Fischer Verlag, Stuttgart (1978)).

This curative and cytostatic effect is with the effect of the well-known Medicinal products cyclophosphamide (EndoxanR) and ifosfamide (HoloxanR) comparable.

The lowest, curative or cytostatic effective dose in the animal experiments given above, for example, 0.01 mg / kg orally 0.01 mg / kg intraperitoneally 0.01 mg / kg intravenously As a general dose range for the curative and cytostatic effect (animal experiments as above) comes for example in question: 0.01 - 100 mg / kg orally, in particular 0.1-10.0 mg / kg 0.01 - 100 mg / kg intraperitoneally, in particular 0.1-10.0 mg / kg 0.01-100 mg / kg intravenously, in particular 0.1-10.0 mg / kg Indications for which the compounds according to the invention can be considered: Malignant diseases of humans and animals.

The pharmaceutical preparations generally contain between 1 mg to 1 g, preferably 10 to 300 mg, of the active component according to the invention (n).

Administration can, for example, in the form of tablets, capsules, Pills, coated tablets, suppositories, ointments, jellies, creams or can be made in liquid form. The following liquid forms of use are, for example: oily or alcoholic or aqueous solutions as well as suspensions and emulsions.

Preferred forms of use are tablets that are between 10 and 200 mg or solutions containing between 0.1 to 5% of the active substance.

The single dose of the active components according to the invention can, for example a) for oral drug forms between 1-100 mg / kg, preferably 10-60 mg / kg b) in the case of parenteral dosage forms (for example intravenous, intramuscular) between 1-100 mq / kg preferably 10-60 mg / kg c) for drug forms for rectal or vaginal Appliw cation between 1-100 mg / kg, preferably 10-60 mg / kg d) for drug forms for local application to the skin and mucous membranes (for example in the form of Solutions, lotions, emulsions, ointments, etc.) between 1-100 mg / kg preferably 10-60 mg / kg - (The doses are based on the free base) examples 1-3 times a day 1 to 10 tablets with a content of 10 to 300 mg more effective Substance or, for example, with intravenous injection 1 to 2 times a day or Several ampoules of 1 to 10 ml content with 10 to 250 mc3 substance are recommended. In the case of oral administration, the minimum daily dose is, for example, 200; the maximum daily dose for oral administration should not exceed 5000. Even a continuous infusion of corresponding doses over 12 and more hours can be used in individual cases recommended.

For the treatment of dogs and cats, the single oral dose is recommended generally between about 10 and 60 mg / kg body weight; the parenteral Dose approximately between 10 and 60 mg / kg body weight.

For the treatment of horses and cattle, the single oral dose is recommended generally between about 10 and 60 mg / kg; the parenteral single dose approximately between 10 and 60 mg / kg body weight.

The dosages indicated on this page and on the previous pages also relate to the use of the compounds according to the invention for immunosuppression.

The acute toxicity of the compounds according to the invention in the mouse (expressed by the LD 50 mg / kg; Miller and Tainter method: Proc. Soc. Exper. Biol. A. Med.

57 (1944) 261) lies between, for example, in the case of oral administration 100 and 1000 mg / kg or above 1000 mg / kg.

The drugs can be used in human medicine, veterinary medicine as well as in agriculture alone or in admixture with other pharmacologically active substances Substances are used.

The following examples serve to further illustrate the present Invention without being limited to this.

Example 1 2- [2- (bis- (2-chloroethyl) -amino-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-thio] -ethanesulfonic acid cis -cyclohexylammonium salt 5.6 g (20 mmol) of 4-hydroxycyclophosphamide (i.e. 2- (bis- (2-chloroethyl) -amino) -4-hydroxy-tetrahydro-2H-1, D, 2-oxaza = phosphorine-2-oxide) and 4.8 g (20 mmol) cyclohexylammonium-2-mercaptoethanesulfonate were dissolved in 100 ml of distilled water, and a little trichloroacetic acid was added and left to stand for 3 days at 0 ° C in the refrigerator. After that, the solvent removed in a high vacuum, the residue taken up in acetone, concentrated again, off Acetone crystallized and isopropanol recrystallized.

Yield: 7.2 g (72% of theory), melting point: 149-151 ° C. (cis form).

Example 2 2- [2- (bis- (2-chloroethyl) -amino-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-thio] -ethanesulfonic acid cis -cyclohexylammonium salt 2.9 g (10 mmol) 2- (bis- (2-chloroethyl) -amino) -4-methoxytetrahydro-2H-1,3,2-oxazaphosphorine-2-oxide and 2.4 g (10 mmol) of cyclohexylammonium 2-mercaptoethanesulfonate were combined with a little trichloroacetic acid dissolved in 10 ml of dimethylformamide and 20 hours at -250C kept.

After a further 3 hours of standing at 0 ° C, with ether until the beginning Turbidity added, rubbed, the crystals after standing for 20 hours at 0 ° C vacuumed, washed and dried.

Yield: 4.7 g (94% of theory), melting point: 1450C (decomp.) Recrystallization from alcohol / ether, m.p .: 149-1510C (cis form).

Example 3 2-z2- (Bis- (2-chloroethyl) -amino) -2-oxo-tetrahydro-2H-1,3s2-oxazaphosphorin-4-yl-thio] -ethanesulfonic acid cis-Cyclohexylammonium salt 1.4 g (5 mmol) 4-Hydroxycyclophosphamid and 1.2 g (5th rnrqol) Cyclohexylammonium-2-mercaptoethanesulfonate were together with a trace Trichloroacetic acid was dissolved in 25 ml of methanol and placed in the refrigerator overnight at -25 ° C ditched.

The reaction solution was then concentrated to about 5 ml, ether Added until the beginning of cloudiness and rubbed. The crystals were after Sucked off, washed and dried after standing at 0 ° C for 20 hours.

Yield: 2.1 g (84% of theory), melting point: 143-145 ° C. (cis form).

Example 4 2- [2- (bis- (2-chloroethyl) -amino-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-thio] -ethanesulfonic acid cis ammonium salt 2.8 g (10 mmol) 4-hydroxycyclophosphamide and 1.6 g (10 mmol) ammonium 2-mercaptoethanesulfonate were combined with a catalytic amount of trichloroacetic acid in 50 Iil water dissolved and left to stand in the refrigerator at OoC for 3 days. After that the water became distilled off in a high vacuum, the residue taken up twice in acetone and dissolved in Reduced vacuum. Recrystallization from Accton / Ether.

Yield: 3.9 (93% of theory), melting point: 131-1330C (cis form).

Example 5 2- [2- (Bis- (2-chloroethyl) -amino-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-thio] -ethanesulfonic acid cis ammonium salt 720 mg (1.8 mmol) 2- [2- (bis- (2-chloroethyl) -amino) -2-oxotetrahydro-2H-1,3,2-oxazaphosphorin-4-yl] -thioethanesulfonic acid were dissolved in a little water, neutralized with ammonia and treated with 4 ml of acetone. The reaction solution stood at -25 ° C. overnight. The crystals were filtered off with suction and recrystallized from methanol / acetone.

Yield: 530 mg (71% of theory), melting point: 133-134 ° C. (cis form).

Example 6 cis-2- / 2- (bis- (2-chloroethyl) -amino) -2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-thio] -ethanesulfonic acid Sodium salt 2.8 g (10 mmol) of 4-Hydroxycyclophosphamid in 10 ml of acetone were with 1.3 g (8 mmol) of sodium 2-mercaptoethanesulfonate in 2 ml of water are added and 3 hours stored at 00 C. The reaction mixture was then in vacuo at 250 C concentrated, the residue taken up in dry acetone and in the cold with dry ether added until the onset of turbidity. When concentrating the solution in A precipitate formed in vacuo.

Yield: 1.5 g (44% of theory) of the cis isomer, m.p .: 83-850 C.

Recrystallization in acetone gave a sparingly soluble in acetone Product.

Example 7 Preparations containing 50 mg or 200 mg of substance as active ingredient 6 according to the following overview: Substance Example 6 50 mg 200 mg mannitol 230 mg 380 mg water for injection purposes ad 2 ml ad 4 ml The substance Example 6 and Mannitol are in each case under gassing with nitrogen and light protection in as much water dissolved so that a solution of 2 ml or 4 ml volume is formed. In the preparation of and further processing is done in such a way that the temperature of the solution does not reach 0.degree exceeds. The solutions are subjected to sterile filtration in a known manner, under aseptic conditions in brown 10 ml injection bottles of 2 ml or 4 ml dosed, fitted with a freeze-drying stopper and placed in a freeze-drying system lyophilized. The system is then aerated with dry nitrogen and the ampoule bottles closed in the system.

The residual water content of the bottle content must not exceed 0.5%.

For the preparation of the injectable solution, the contents the bottles with 50 mg of active ingredient in 5 ml, with 200 mg of active ingredient in 10 ml of water for Injections resolved.

Claims (9)

Claims cis-Oxazaphosphorin-4-thio-alkanesulfonic acids and their neutral salts of the general formula I. where R1, R2 and R3, which can be identical or different from one another, represent hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl and at least two of these radicals are 2-chloroethyl and / or 2-methanesulfonyloxyethyl, R4 is hydrogen or methyl is, a straight or branched chain C2-6-alkylene, and the hydrogen cation, an alkali or alkaline earth cation, the guanidinium, morpholinium or cyclohexylammonium cation or the cation which is derived from an amine of the formula NR5R6R7, in which the radicals R5 to R7 are identical or different and denote hydrogen, C1 ~ C2-alkyl groups or oxyethyl groups, or in which Y + is the ethylenediammonium cation H3N + -CH2CH2-N + H3 or the piperazonium cation and z is 1 if is a monobasic cation, or 2 if is a dibasic cation or the cation of a compound having two monobasic cations. 2. Compounds according to claim 1, characterized in that in formula I Y + the hydrogen cation, the guanidinium, morpholinium or cyclohexylammonium cation or the cation derived from an amine of the formula NR5R6R7, wherein the chain R5 to R7 are identical or different and are hydrogen, C1-C2-alkyl groups or O>: yethyl groups, or in which Y + is the ethylenediammonium cation H3N + -CH2CH2-N + H3 or that is the piperazonium cation and z is 1 when + Y is a monobasic cation is, or 2 when Y + is basic cation or the cation of a compound with two monobasic cations. 3. Compounds according to claim 1, characterized in that in formula 1 is an Ealium- or Flidalkalication. 4 compounds according to claim 1 or 2, characterized in that in formula X YO is the cyclohexyl ammonium cation. 5. Compounds according to claim 1 or 2, characterized in that in formula I Y + is the guanidinium cation. 6. 2- [2- (Bis- (2-chloroethyl) -amino) -2-oxotetrahydro-2H-1,3,2-oxazaphosphorin-4-yl] -thioethanesulfonic acid cis sodium salt. 7. A process for the preparation of the cis-oxazaphosphorin-4-thio-alkanesulfonic acids and their neutral salts according to one or more of claims 1 to 6 by reacting a 4-hydroxy- or 4-C1 ~ 4-alkoxy-oxazaphosphorin of the general formula II in which R1, R2, R3 and R4 have the same meaning as in formula I and Z is hydrogen or C1 ~ 4-alkyl, with a compound of the general formula III HS-X-SO3-Yü III in which X and Y + have the same meaning as in formula I, and, if Y + is hydrogen and neutralization of the oxazaphosphorin-4-thio-alkanesulfonic acids obtained with a base corresponding to the other meaning of Y +, neutralized according to DE-PS ..... (patent application PP 31 11 428.8 thereby characterized in that the reaction is carried out in an anhydrous solvent or mixture of several such solvents or in the presence of only a small proportion of water. 8. The method according to claim 7, characterized in that the Reaction between compounds II and III at a temperature between -30 and +200 C. 9. Pharmaceutical preparations containing a compound as active ingredient according to claims 1 to 6 in addition to the usual physiologically compatible carriers and / or diluents.