JPH029599B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel 4-carbamoyloxy-oxazaphosphorine compounds and methods for their preparation as well as pharmaceutical formulations containing them as active substances. 2-[bis-(2-chloroethyl)-amino]-2-oxo-tetrahydro-, a cytostatic agent known from German Patent Application No. 2231311
2H-1,3,2-oxazaphosphorine (cyclophosphamide), 3-(2-chloroethylamino)
-2- [Bis-(2'-chloroethyl)-amino]
-2-oxo-tetrahydro-2H-1,3,2
-Oxazaphosphorine (trophosphamide), 3
-(2-chloroethylamino)-2-(2'-chloroethylamino)-2-oxo-tetrahydro-
2H-1,3,2-oxazaphosphorine (ifosfamide), 3-(2-chloroethylamino)-
2-(2'-methanesulfonylamino)-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorine (sphosphamide) and other similar cyclophosphamides with a hydroperoxy group -OOH at the 4-position. It is known that when introduced, a compound having a useful cell proliferation inhibitory effect can be obtained. However, these compounds have extremely low stability and cannot be converted into the pharmaceutical agents necessary for their use in human therapy. It is therefore an object of the present invention to provide novel cyclophosphamide compounds substituted in the 4-position thereof by another converted hydroxy group, which are characterized by particularly high cytostatic activity and improved stability. That's true. The novel compounds of the present invention have the formula {However, in the formula, Z is a group

[Formula] or a group -OR ₇ , X is oxygen or sulfur, and R ₁ , R ₂ and R ₃ are the same or different and represent hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl; , the radicals R ₄ are the same or different and represent hydrogen, methyl or ethyl,
R ₅ and R ₆ are the same or different, hydrogen, C _1-4
-Alkyl, hydroxy-C _1-4 - represents alkyl or phenyl, R ₇ is hydrogen, carbamoyl group,
-OR ₈ (In the formula, R ₈ is hydrogen, C _1-4 - alkyl,
phenyl or benzyl), straight-chain or branched C _1-18 -alkyl [possibly the same or different, hydroxy, halogen, -COOH,
-COOR ₉ , -CONH ₂ , -phenyl, benzyloxycarbonyl, -N(R ₉ ) ₂ , -N(R ₉ ) ₃ , -OR ₉ , -SR ₉ , -SO-R ₉ , -SO ₂ -R ₉ , _-SO3H or -PO( _CH3 ) ₂ (in the formula, _R9 represents methyl or ethyl) -
phenyl-C _1-4 -alkyl (optionally substituted with phenyl and/or
or substituted with one or two carboxy groups on the alkyl moiety), allyl,
C _3-8 - cycloalkyl, tetrahydrofuranyl,
Tetrahydropyranyl, phenyl (optionally 1
species or two of C _1-4 -alkyl, C _1-2 -alkoxy, nitro, halogen, trifluoromethyl, -
SO ₂ TH ₂ , substituted by carboxy, benzyloxycarbonyl and/or carbo-C _1-4 -alkoxy), benzyl, benzhydryl,
naphthyl, fluorenyl, pyridyl, thienylbenzoyl or C _1-4 -alkanoyl, or R ₅ and R ₆ or R ₆ and R ₇ taken together with the atoms to which they are attached, optionally an oxygen atom, C _1-4 - form a saturated heterocycle containing a nitrogen atom substituted with alkyl or a -S-, -SO- or -SO ₂ - group, or R ₆ and R ₇
together with adjacent nitrogens to form an aziridine ring optionally substituted by a cyano or carbamoyl group, and pharmaceutically useful salts thereof. Compounds of the formula that are preferred because they have particularly favorable properties and are easy to prepare are Z
but

[Formula], X is oxygen, R ₅ and R ₆ are the same or different and are hydrogen, methyl or ethyl, and R ₇ is hydrogen, linear or branched C _1-18 - A compound when it is alkyl, phenyl or benyl. Particularly preferred compounds are those in which all R ₄ groups in the formula represent hydrogen atoms among the preferred compounds mentioned above. Another preferred group of compounds is in which X is oxygen, R ₁ , R ₂ and R ₃ are the same or different and represent hydrogen or a 2-chloroethyl group, R ₄ , R ₅ and R ₆ represent hydrogen, and R ₇ is hydrogen, benzyl, phenyl (optionally substituted by 1 or 2 carboxy groups), C _1-4 -alkyl (optionally substituted by 1 carboxy group) or phenyl-C _{1 ~4} -Alkyl (optionally substituted by one or two carboxy groups in the phenyl and/or alkyl part of the group). According to the present invention, 4-carbamoyloxy-oxazaphosphorine of the formula (wherein R ₁ , R ₂ , R ₃ and R ₄ are as defined in the formula and Y is hydrogen, methyl or ethyl) optionally heated in the presence of an inert solvent. or with cooling and/or in the presence of an acid catalyst. (wherein Z, R ₅ and X are as defined in the formula). water, lower alkyl halides such as methylene chloride, lower alkyl ketones such as acetone,
Diethyl ether, dimethylformamide (DMF), hexamethylphosphoric triamide (HMPT) or similar solvents or mixtures of such solvents are suitable inert solvents for use in the above process. The reaction starts from -35℃ to +
It can be carried out at temperatures in the range up to 50° C., ie optionally with cooling, at room temperature or with heating. The reaction is carried out with acid catalysts such as inorganic acids, trichloroacetic acid, trifluoromethanesulfonic acid or Lewis acids such as aluminum chloride,
It can be carried out in the presence of zinc chloride or titanium tetrachloride. Another embodiment of the method for producing a compound of the formula, wherein Z is -NR ₆ R ₇ (wherein R ₆ is hydrogen) and R ₅ is hydrogen, is a compound of the formula (wherein R ₁ to _{R 4} are as defined in the formula) in an inert solvent, -
characterized in that it is subjected to reaction with a compound of _{the formula} . The course of the reaction in both embodiments can be followed by thin layer chromatography. Isolation of the thin-layer chromatographically homogeneous material is carried out by conventional preparation methods for such products, in particular by crystallization or chromatographic purification. The structure can be confirmed by melting point, thin layer chromatography,
This can be done by elemental analysis, infrared and/or ¹ H-NMR spectroscopy. The compounds of the formula and used as starting materials in the process according to the invention are widely known and can be used in crystalline form or as crude products. They can be synthesized by known methods, for example as follows. 4-Hydroxy-oxazaphosphorine can be obtained by reduction of 4-hydroperoxy derivatives [for example, "J.Med.Chem." Vol. 18, p. 376 (1975)
reference〕. 4-Methoxy- or 4-ethoxy-oxazaphosphorines can be produced from 4-hydroxy derivatives in methanol or ethanol or inert solvents containing methanol or ethanol in the presence of acid catalysts. Hydroxyurea derivatives are produced by converting appropriately substituted isocyanates or carbamic acid chlorides with hydroxylamine or N-monosubstituted hydroxylamines. Racemic cis- and trans-isomers can be produced from 4-carbamoyloxy-oxazaphosphorine according to the invention. The cis configuration is
2R, 4R/2S, 4S, but the other transformer arrangement is
2R, 4S/2S, 4R, and the relationship is according to IUPAC nomenclature conventions and consistent with the literature on corresponding oxazaphosphorine derivatives. The cis or trans form can be generated intentionally by selecting the reaction conditions. Pharmacologically, the isomers do not show significant differences. The compounds according to the invention have particularly valuable chemotherapeutic properties. Previously known cyclic phosphamides such as cyclophosphamide (cyclo-
phosphamide), trofosfamide, ifosfamide
They exhibit substantially equivalent cancer toxicity and chemotherapeutic efficacy against experimentally implanted tumors in rats. They have a direct alkylating effect in aqueous solution, unlike cyclic phosphamides, and have high cytotoxicity in vitro. In the latter case, for example, cyclophosphamide requires enzymatic activation and has virtually no cytotoxic effect in vitro. The acute toxicity of the compounds according to the invention is considerably lower than that of known cyclic phosphamides. For example, it is about 4 times lower than that of the reference substance cyclophosphamide, thus the therapeutic ratio of the compounds of the invention is considerably improved. Alternatively, the compounds according to the invention have distinct advantages over the prior art cyclic phosphamides with regard to organotoxic side effects such as leukocyte depletion and immunosuppression. The 4-carbamoyloxy-oxazaphosphorines according to the invention are useful in treating human malignancies and similar malignancies such as leukemia. They are administered in doses ranging from 0.01 to 100 mg per kg body weight per day. The pharmaceutical formulations used in such treatments are those commonly used for cyclic phosphamides and other known cytostatic agents, oxazaphosphorine. They can be produced in conventional manner using conventional additives, diluents and/or carrier materials. The compounds of the invention can be administered to animals, including humans, in a manner similar to that known in connection with the cyclic phosphamides described above, and for similar disease states. However, given the lower toxicity of the compounds of the invention, they can be used at higher dosage ratios and thus have a significantly expanded effective therapeutic dose range. Ordinarily, the compounds of the invention will be administered by conventional routes, such as orally or by injection, as conventional formulations prepared by mixing them with physiologically acceptable excipients and/or diluents. be done. The production of such formulations and the methods by which they are administered are well known to those skilled in the art. The compounds according to the invention are useful in preparing pharmaceutical products or medicaments containing one or more such compounds as active agents, optionally together with other pharmacologically or pharmaceutically active agents. It is. The production of the pharmaceutical preparations is carried out in a manner known per se using known and customary pharmaceutical excipients or customary carrier substances or diluents. Useful as carriers and excipients are, for example, ``ULLMANNS ENCYKLOPAEDIE DER''.
TECHNISCHEN CHEMIE, Vol. 4, pp. 1-39 (1953), J.Pharm.Sciences, Vol. 52, pp. 918 et seq. (1963), HVCzetsch-Lindenwald (ed.), Hilfsstoffe fu¨r Pharmazie und angrenzende.
Gebiete, Pharm.Ind.” Volume 2, pp. 72 et seq. (1961
), HP Fiedler, “Lexikon der
Hilfsstoffe fu¨r Pharmazie, Kosmetik und
This is a compound described in "Gebiete" (1971). Examples include gelatin, natural sugars such as sucrose or lactose, lecithin, pectin, starches (e.g. corn starch), alginic acid, tylose, talc, stone pine, silicic acid (e.g. colloidal silicic acid), cellulose, cellulose derivatives (e.g. cellulose ethers or those whose hydroxy groups are partially etherified with saturated lower aliphatic alcohols and/or with saturated lower aliphatic oxyalcohols (e.g. methyloxypropyl cellulose), stearates, having 12 to 22 carbon atoms fatty acids, especially magnesium and calcium salts of saturated fatty acids (e.g. stearates), emulsifiers, oils and fats, especially vegetable fats (e.g. peanut oil, castor oil, olive oil, sesame oil,
cottonseed oil, corn oil, wheat oil, sunflower oil,
cod liver oil, mono-, di-, and tri-glycerides of saturated fatty acids C ₁₂ H ₂₄ O ₂ to C ₁₈ H _{36 O} 2 and mixtures thereof), pharmaceutically compatible mono- or polyols and polyglycols such as polyethylene. Glycols and their derivatives, aliphatic monoalcohols (having 1 to 20 carbon atoms) of aliphatic saturated or unsaturated fatty acids (having 2 to 22, especially 10 to 18 carbon atoms), or polyols such as glycols , esters with glycerol, diethylene glycol, pentaerythritol, sorbitol, mannitol, and the like, as optionally esterified, benzyl benzoate, dioxolane, glycerol formal, tetrahydrofurfuryl alcohol, C ₁
~C ₁₂ - polyglycol ether with alcohol,
dimethylacetamide, lactamide, lactate,
Mention may be made of ethyl carbonate, silicones (especially dimethylpolysiloxane with medium viscosity), magnesium carbonate and the like. To prepare the solution, for example water or a physiologically compatible organic solvent such as ethanol,
1,2-propylene glycol, polyglycols and their derivatives, dimethyl sulfoxide, fatty alcohols, triglycerides, partial esters of glycerol, paraffin and their analogs are used. Customary solubilizers or emulsifiers known in the manufacture of pharmaceutical products, such as polyvinylpyrrolidone, fatty acid esters of sorbitan, such as sorbitan trioleate, lecithin, gum arabic, tragacanth, polyoxyethyl sorbitan monooleate, polyoxyethyl polyoxyethylated oleotriglycerides, linoleated oleotriglycerides,
Polyethylene oxide condensation products of fatty alcohols, alkylphenols or fatty acids or 1-
Methyl-3-(2-hydroxyethyl)-imidazolidone-(2) can be used. As used above, the term "polyoxyethylated" means that each compound generally has between 2 and 40
In particular, it means containing polyoxyethylene chains with a degree of polymerization of 10 to 20. Such polyoxyethylated products can be prepared, for example, by converting the corresponding hydroxy compound (for example a mono- or diglyceride or an unsaturated compound such as one containing an unsaturated oleyl group) into ethylene oxide (for example per mole of glyceride).
40 moles of ethylene oxide). Examples of oleotriglycerides are olive oil, peanut oil, castor oil, sesame oil, cottonseed oil, and corn oil [see HP Fiedler, Lexikon
191-195 (1971
2007)]. Furthermore, the pharmaceutical preparations according to the invention contain preservatives, stabilizers, buffers such as calcium hydrogen phosphate, colloidal aluminum hydroxide, flavorings, antioxidants and complexing agents (e.g. ethylenediaminetetraacetic acid) and the like. can do. Where possible, the active agent may be stabilized using physiologically compatible acids or buffers to stabilize it.
Adjust the pH to about 3-7. nearly neutral or slightly acidic
PH (up to PH5) is generally preferred. Common antioxidants are, for example, isomeric sodium bisulfite, ascorbic acid, gallic acid, gallic acid alkyl esters, butylhydroxyanisole, nordihydroguaiaric acid, tocopherols and tocopherols and synergistically active reagents. (Reagents that bind heavy metal cations by complex formation, e.g. lecithin, ascorbic acid, phosphoric acid)
It is a combination of Addition of synergistically active reagents considerably increases the antioxidant activity of tocopherols. Useful preservatives are, for example, sorbic acid, P-hydroxybenzoic acid esters (eg lower alkyl esters), benzoic acid, sodium benzoate, trichloroisobutyl alcohol, phenols, cresols, benzethonium chloride and formaldehyde derivatives. The conversion of the compounds according to the invention into pharmaceutical products is carried out applying customary galenic components and customary methods. For example, active agents or reagents and excipients and/or carrier materials are typically
Mix thoroughly (for example using a conventional mixer) while applying a temperature of preferably 20 to 50°C, especially room temperature. Further details can be found in “Pharamzeutische
Technology” (1978). The compounds according to the invention and the pharmaceutical preparations containing them may be applied to the skin, to the mucous membranes, or for example orally, enterally, to the pulmonary artery, respectively.
Rectally, through the nose, vagina, or tongue, intravenously, intraarterially, intracardially, intramuscularly, intraperitoneally, intradermally, subcutaneously, It is administered intrapleurally or intrathecally, and generally intrasinally. In view of the very favorable results, the active compounds according to the invention can be used as medicaments in combination with other active compounds, in particular uroprotective agents, such as, for example and preferably, sodium-2-mercapto-ethanesulfonate or 2,2'-dithiodi-(ethane). Sulfonic acid)
disodium salt], but also with other systemic or local antidotes. The compounds according to the invention exhibit good cytostatic and curative activity when applied intravenously, intraperitoneally or orally to rats and mice bearing various experimental tumors. For example, depending on the dose, therapeutic activity is achieved with the compounds according to the invention. That is, it is achieved after intraperitoneal implantation of Yoshida ascites sarcoma AH13 cells and, one day later, by administering them at various doses intravenously, intraperitoneally or orally.
For example, tumor ascites fluid is collected under aseptic conditions (as evidenced by bacteriological control) from 5-7 day old rats with Yoshida ascites sarcoma. The average cell number of ascites is 2×10 ⁵ cells per μ. The cell number is brought to 10 ⁸ cells per ml by adding Tyrode's solution. 0.4 ml (4 x 10 ⁷ cells) thereof is transplanted intraperitoneally into the test animal. In untreated control animals the rate at which these tumors begin to grow is 95% and the average time to death is 8 days (e=±0.5d). Again, the compound to be tested is obtained by transplanting tumors and
Administered intraperitoneally after hours. Six animals are used in each test group. Regarding the criteria for curative activity, cure is defined as no recurrence or metastasis observed for 90 days after treatment. After determining the cure rate (%), the dose that causes cure in 50% of the animals (DC50) is calculated from dose-active radiation using probit analysis by R. Fischer. This DC50 is the value of tumor-infected animals.
The dose is 50% curative. The DC50 of the compounds of the present invention tested by the above method is shown in the third column of Table 1 below. In the above test method by intraperitoneal administration, the dose for the compounds of the invention is, for example, 0.05 to 30 mg/Kg in rats. For example, when using a dose corresponding to the DC50, leukemia L1210 was observed in mice.
can increase the average survival time by 100%. In order to determine cytotoxic activity in vitro, "Naturwissenschaften" Vol. 43, p. 199 (1956
Yoshida ascites sarcoma cells, freshly harvested as described in 2010), are incubated with increasing concentrations of the test compound in Ringer's solution for 2 hours at 37°C. After washing away the test compound, the potential for implantation of tumor cells into untreated test animals is determined. Quantitative values for cytotoxic activity in vitro are obtained by determining the concentration at which survival of implanted tumor cells is inhibited in half of the animals, ie, the EC50. When using the compounds of the present invention, the above tests can be carried out in vitro, for example, from 1 to
Shows cytotoxic activity at a concentration of 100 μg/ml. To test the activity of the compounds of the invention against Leukemia L5222 in rats, blood from donor rats of the BD strain is removed by cardiac puncture under pentobarbital anesthesia (intraperitoneal application). The number of white blood cells in the blood is 50000 cells and 150000 cells per μ
The blood was collected using a sterile sodium chloride solution varying between cells with a white blood cell count of 5 x ¹⁰³ cells/μ.
dilute to 1 ml (=5×10 ⁶ cells) is intraperitoneally transplanted into a test animal of the same species. The tumor growth rate is 100%, the time to death of the test animals is 8-10 days, and 6 animals are used for each test group. Test compounds are administered 5 days after leukemia transplantation. The criteria for determining curative efficacy is no recurrence for 90 days after treatment. R.Fischer after determining the cure rate (%)
The DC50 dose can be determined from the dose-active line using probit analysis by Dr. This is the dose that produces cure in 50% of the test animals. This effect against Leukemia L5222 in rats when the compound according to the invention was administered intraperitoneally.
The DC50 is, for example, in the range of 0.5 to 20 mg per kg of rat. Furthermore, cytostatic activity is obtained when the compound according to the present invention is administered once or several times (four times) at different doses for several days after Leukemia L1210 10 ⁶ cells are implanted into the peritoneal cavity of a mouse. Cytostatic activity is expressed as an increase in the mean survival time of tumorous animals, a dose-dependent percentage increase in survival time greater than that of unaffected controls. This test method is described, for example, in N. Brock's “Pharamkologische Grundlagen der Krebs”.
"Chemotherapie" [edited by A. Georgii "Verhandlungen der Deutschen
"Krebsgesellschaft" Volume 1, pp. 15-42 (1978
year)]. This curative and cytostatic activity can be compared to that of the known cytostatic agents cyclophosphamide and ifosfamide. Furthermore, the compounds according to the invention exhibit a good healing width. Furthermore, they have distinct advantages over the known cytostatic agents cyclophosphamides, taking into account organotoxic side effects such as a reduction in white blood cell counts and a weaker immunosuppressive effect. Furthermore, the urinary toxicity of the compounds according to the invention is considerably low. Furthermore, it can be avoided by prophylactically administering uroprotective agents such as and preferably sodium 2-mercapto-ethanesulfonate. The lowest doses that already showed curative or cytostatic effects in the above animal tests are, for example, as follows. 0.01mg/Kg for oral administration, 0.01mg/Kg for intraperitoneal administration, 0.01mg/Kg for intravenous administration. Typical dosage ranges for therapeutic and cytostatic activity (in animal studies similar to those described above) are, for example: Oral administration: 0.01-100mg/Kg, especially 0.1-100mg/Kg
10.0mg/Kg, 0.01-100mg/Kg for intraperitoneal administration, especially 0.1-100mg/Kg
10.0mg/Kg. For intravenous administration: 0.01-100mg/Kg, especially 0.1-100mg/Kg
10.0mg/Kg. Compounds according to the invention are useful in curing malignant diseases in humans and animals. Toxicity tests for the compounds of the present invention were conducted as follows. Determination of lethal dose in a single dose (acute toxicity). The test compound was suspended in a 0.5% tragacanth suspension and administered intraperitoneally to male rats (Sprague-Dawley strain) with an average body weight of 160 g.
The maximum dose was 1000mg/Kg. The dose administered was varied in a logarithmic series with a factor of 2.15. Experimental animals were observed for 30 days, and the toxicity was completely consistent with known nitrogen mustard toxicity, which included hair loss, mucous membrane inflammation, tooth decay, apathy, and diarrhea. Most deaths occurred by the seventh day of observation. Miller and
Approximately the mean lethal dose (LD50) was determined from the graph according to Tainter's method. The results are shown in the second column of Table 1.

【table】

[Table] Generally, pharmaceutical preparations are 1 mg to 1 g, preferably 10
mg/300 mg of active compound or compound according to the invention. Administration can be carried out, for example, as tablets, capsules, pills, dragees, suppositories, ointments, jellies, creams or in liquid form. Liquid application forms are, for example, oily solutions or solutions in alcohol or water and suspensions and emulsions. Preferred dosage form is 10-200mg
tablets containing 0.1-5% of active compound or solutions containing 0.1-5% of active compound. A single dose of the compound according to the invention is, for example, as follows. (a) 1 to 100 mg/Kg, preferably 10 to 60 mg/Kg, for pharmaceutical preparations to be administered orally; (b) for pharmaceutical preparations to be administered parenterally (e.g. intravenously or intramuscularly); 1 to 100 mg/Kg, preferably 10 to 60 mg/Kg for pharmaceutical preparations)
Kg, (c) 1 to 100 mg/Kg for pharmaceutical preparations for rectal or vaginal application, preferably 10 to 60 mg/Kg, (d) Pharmaceutical preparations for topical application to the skin or mucous membranes, e.g. 1 to 1 for solutions, lotions, emulsions, ointments or similar forms)
100mg/Kg, preferably 10-60mg/Kg. (The above single doses are stated for the active compound in free base form). For example, 1 to 10 tablets each containing 10 to 300 mg of active compound can be administered 1 to 3 times per day. For intravenous injection, ampoules containing 10-50 mg of active compound per day (1-10
ml) can be administered once or twice. The minimum daily dose for oral administration is, for example, 200 mg, and the maximum daily dose for oral administration should not be more than 500 mg. The compounds can also be administered by continuous intravenous infusion over a period of 12 hours or more in special cases. When treating dogs and cats, a single oral dose is generally about 10-60 mg/kg body weight. Parenteral dosage is approximately 10-60 mg/kg body weight. When treating horses and cattle, a single oral dose is generally about 10-60 mg/Kg. A single parenteral dose is approximately 10-60 mg/kg body weight. Acute toxicity of the compounds of the present invention in mice [expressed in LD50mg/Kg, "Proc.Soc.Exper.
Biol.a.Med., Vol. 57, p. 261 (1944)], for example, 100~
1000mg/Kg or more than 1000mg/Kg respectively. The pharmaceutical preparations according to the invention can be used alone or in admixture with other pharmacologically active agents in human therapy, in veterinary therapy or agriculturally. EXAMPLES In order to better understand the present invention, examples will be described below, but the present invention is not limited thereto. Example 1 2-[bis-(2-chloroethyl)-amino]
-2-oxo-tetrahydro-2H-1,3,
2-oxazaphosphorin-4-yl-oxy-
Urea 4-hydroxycyclophosphamide (i.e. 2-[bis-(2-chloroethyl)-amino]-
4-Hydroxytetrahydro-2H-1,3,2
-oxazaphosphorine-2-oxide) 15g (54
mmol) and 4.4 g (58 mmol) of hydroxyurea were dissolved in 70 ml of DMF, acidified with trichloroacetic acid (PH 3-4), and stored at 0 °C in the refrigerator for 20 min.
Leave it for a while. The crystalline sludge obtained is diluted with 70 ml of ethyl acetate, separated off with suction after 2 hours, washed, dried and recrystallized from methanol. Yield cis form 11.3g (62% of theoretical value), m.
p.139-143℃ (decomposition). Example 2 2-[bis-(2-chloroethyl)-amino]
-2-oxo-tetrahydro-2H-1,3,
2-oxazaphosphorin-4-yl-oxy-
Urea (cis form) 4-hydroxycyclophosphamide 1.1g (4
mmol) in methanol, treated with traces of trichloroacetic acid, left overnight at −25°C, then methanol was gently distilled off, the residue was dissolved in a small amount of methylene chloride, dried, and concentrated. Then 4-methoxycyclophosphamide (i.e. 2
―[Bis-(2-chloroethyl)amino]-4-
1.2 g of methoxy-tetrahydro-2H-1,3,2-oxazaphosphorine-2-oxide are obtained. Dissolve 1.2 g of 4-methoxycyclophosphamide and 304 mg of hydroxyurea in 3 ml of DMF,
Then, keep it in the refrigerator at -25℃ for 20 hours. The crystalline sludge is diluted with 3 ml of ethyl acetate, separated with suction, washed, dried and recrystallized from methanol. Yield, same product as Example 1 670
mg (50% of logical value). Example 3 2-[bis-(2-chloroethyl)-amino]
-2-oxo-tetrahydro-2H-1,3,
2-oxazaphosphorin-4-yl-oxy-
Urea (trans form) 4-(Hydroxycyclophosphamide 16g (58
5.2 g (68 mmol) of hydroxyurea are dissolved in 160 ml of water, acidified (PH 3-4) with trichloroacetic acid and left in the refrigerator at 0° C. for 20 hours. The crystalline sludge is then separated by suction, washed with water, dried over phosphorus pentoxide under high vacuum and recrystallized from methanol/chloroform. Yield, trans form of the product of Example 1 12.7
g (65% of theory), mp 148°C (decomposition). Example 4 3-(2-chloroethyl)-2-[bis-
(2'-chloroethyl)-amino]-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-oxy-urea 4-hydroxytrophosphamide (i.e. 3
-(2-chloroethyl)-2-[bis-(2'-chloroethyl)-amino]-4-hydroxytetrahydro-2H-1,3,2-oxazaphosphorine-2-oxide) 20 g (50 mmol) and 5.3 g (70 mmol) of hydroxyurea are dissolved in 100 ml of DMF and cooled to -15°C. Next, make it acidic (PH3-4) with trichloroacetic acid and 55% at -15℃.
Stir for hours. After standing overnight at 0°C, the reaction solution is diluted with twice the amount of water. Extraction was then carried out by shaking four times with 300 ml of acetate/methanol (10:1) in each case, the combined acetate phases were washed twice with water, dried over sodium sulfate and concentrated under vacuum to form an oil. 22g is obtained. After treatment with ethyl acetate/methanol, 4.2 g (mp 106-110°C) crystallize out. The mother liquor is fractionated by silica gel column chromatography using chloroform/methanol (10:1) and recrystallized along with the first crystals from ethyl acetate/methanol. Yield, 7.0 g (35% of theoretical value), m.
p.115-116℃ (decomposition). Example 5 3-benzyl-1-[2-(bis-(2-chloroethyl)-amino)-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-oxy]- Urea 540 mg (3.25 mmol) of 3-benzyl-1-hydroxyurea in 40 ml of acetone and a catalytic amount of trichloroacetic acid are added to 900 mg (3.25 mmol) of 4-hydroxycyclophosphamide in 1 ml of methylene chloride. The mixture is kept at -25 DEG C. overnight, then the crystals are separated by suction, washed with acetone and ether and recrystallized from ethyl acetate. Yield 500mg (40% of theoretical value), mp122
~123℃ (decomposition). Example 6 3-(o-bromophenyl)-1-[2-(bis-(2-chloroethyl)-amino)-2-oxotetrahydro-2H-1,3,2-oxazaphosphorin-4-yl- Oxo]-Urea 560 mg (2 mmol) of 4-hydroxycyclophosphamide in 10 ml of acetone are treated with 460 mg of 3-o-bromophenyl-1-hydroxyurea and a catalytic amount of trichloroacetic acid and left at -25°C. After 2 days the crystals are separated by suction and recrystallized from acetone. Yield 320 mg (theoretical value 32
%), mp110-111℃ (decomposition). Example 7 N-[2-(bis(2-chloroethyl)-amino)-2-oxo-tetrahydro-2H-1,
3,2-Oxazaphosphorin-4-yl-oxy]-4-morpholinocarboxamide 1.2 g (4.3 mmol) of 4-hydroxycyclophosphamide in 15 ml of acetone was converted to 630 mg (4.3 mmol) of N-hydroxymorpholino-carboxamide.
and traces of trichloroacetic acid, and −
Hold at 25°C. After 4 days the crystals are separated by suction and recrystallized from acetone. Yield 780mg
(45% of theoretical value) mp123-124℃ (decomposition). Examples 8 to 70 Similarly corresponding 4-hydroxy-tetrahydro-2H-1,3,2-oxazaphosphorine-2-
Starting from the oxide, 4-ureidooxy derivatives having the following general formula are prepared.

【table】

【table】

【table】

【table】

【table】

[Table] They are distinguished by their R _f values obtained. Further distinguishing features are determined in the same manner as for crystalline compounds.
Example 71 Ethyl-2-[bis-(2-chloroethyl)-
Amino]-2-oxo-tetrahydro-2H-
1,3,2-oxazaphosphorin-4-yl-
Oxy-carbamate 4-hydroxycyclophosphamide 550mg (2
210 mg (2 mmol) of ethyl hydroxy carbamate (hydroxyurethane) are dissolved in 5 ml of alcohol-free methylene chloride. Catalytic amount of trichloroacetic acid and molecular sieves
Add 4A to it. The reaction mixture is left at -25°C for 3 days. The molecular sieves are then separated and washed once with a dilute solution of sodium bicarbonate. The methylene chloride phase is dried over sodium sulfate, part of the solvent is evaporated under vacuum and then diluted with ether. After standing at −25° C. for 20 hours, the separated crystals are separated, washed, and dried. Yield 290 mg (40% of theory), mp 96°C. Example 72 Benzyl-2-[bis-(2-chloroethyl)
-Amino]-2-oxo-tetrahydro-2H
-1,3,2-oxazaphosphorin-4-yl-oxy-carbamate 4-hydroxycyclophosphamide 750 mg (2.7
450 mg (2.7 mmol) of benzyl-hydroxycarbamate are dissolved in 6 ml of alcohol-free methylene chloride. A small amount of trichloroacetic acid is added to it and the solution is left in the refrigerator at -25°C for 3 days. The resulting solution is separated and the mother liquor is diluted with 5 ml of chloroform and then with water and then washed with a dilute solution of sodium bicarbonate and water. The washed solution is dried over sodium sulfate and evaporated under vacuum.
The oily residue is then recrystallized from ethyl acetate containing a small amount of methanol. Yield 680mg
(59% of theoretical value), mp112~114℃. Example 73 {3-[2-(bis-(2-chloroethyl)-
Amino)-2-oxo-tetrahydro-2H-
1,3,2-oxazaphosphorin-4-yl-
Oxy[ureido]-acetic acid cyclohexylamine salt (a) 3-hydroxy-ureidoacetic acid glycine benzyl ester hydrochloride 56.5g
(0.28 mol) is suspended in 300 ml of toluene. Dry, gaseous phosgene is introduced with stirring for 2 hours while the reaction mixture is heated on an oil bath heated to 140°C. The reaction mixture is then evaporated under vacuum and the crude benzyl isocyanate acetate residue is distilled under high vacuum. Yield 51g (95% of theoretical value),
bp0.05: 100-102℃. 6.6g hydroxylamine in 200ml dioxane
(0.2 mol) solution of 28.7 g (0.15 mol) of benzyl isocyanate acetate in 50 ml of dioxane.
Add dropwise to the solution with stirring and while cooling briefly. Continue stirring for an additional hour at room temperature. The reaction mixture is then evaporated under vacuum. The resulting residue i.e. crude benzyl
3-Hydroxyureido acetate is recrystallized from ethyl acetate. Yield 28.1g (theoretical value 83.6
%), mp113~120℃. 5 g of palladium-activated carbon are added to a solution of 22.4 g (0.1 mol) of benzyl-3-hydroxyureido acetate in 300 ml of methanol. Hydrogen is introduced while shaking. Hydrogen absorption ends after about 20 minutes. The catalyst is sucked off and the liquid is evaporated under vacuum. The solid residue, crude 3-hydroxyureidoacetic acid, is recrystallized from dioxane. Yield 9.8g (73% of theoretical value), m.
p.135℃. (b) 6.1 g of 4-hydroxycyclophosphamide
(2 mmol) is added to a solution of 2.4 g (18 mmol) of 3-hydroxyureidoacetic acid in 10 ml of water and 25 ml of acetone. This reaction mixture was heated at −25°C.
Leave it overnight. Then 25 ml of acetone and 1.8 g of cyclohexylamine (18
mmol) solution is added to it. After standing for 2 hours, the precipitate is filtered off with suction and recrystallized from acetone containing a small amount of methanol. Yield 3.1
g (44% of theoretical value), mp107-108℃. Example 74 3-[N,N-(bis-(2-chloroethyl)
-Diamino)-phosphinyl-oxy]-propionacedehyde-oxime (aldophosphamide-oxime) 4-hydroperoxycyclophosphamide 4.0
g (13.7 mmol) is suspended in 50 ml of water while cooling on ice. Add 500mg of Na ₂ S ₂ O ₃ ×5H _{2 O} to it. 5
Adjust the pH using a pH measuring device while stirring at ~10°C and maintain the pH at 4.5-5.5 by adding 2N sulfuric acid.
A concentrated solution of sodium thiosulfate is added dropwise thereto until the PH of the reaction mixture no longer increases continuously. Stirring was continued for half an hour at about 10°C and the pH was brought to 5 by addition of 2N sodium hydroxide.
An aqueous solution of 950 mg of hydroxylamine hydrochloride is added dropwise while maintaining . The resulting reaction mixture is left overnight at 5° C. in a condenser. The reaction mixture is then extracted four times with 50 ml each of ethyl acetate, the organic extracts are dried over sodium sulfate and evaporated under vacuum at 30°C. The residue is dissolved in methylene chloride and the crystals that have separated are separated after one day. Yield 3.4g (85% of theory), mp79-81℃. Example 75 3-p-bromo-1-[2-(bis-(2-chloroethyl)-amino)-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-oxy ]-Urea 4 g (20 mmol) of p-bromophenyl isocyanate in 40 ml of acetone and 5.8 g (20 mmol) of aldophosphamide-oxime in 60 ml of acetone.
Add to. The reaction mixture is stirred with cooling for 5 hours. After standing for 2 hours, the separated crystals are separated, dried under vacuum at 40° C. in a rotary evaporator, and recrystallized in methanol. Yield: 8.1g (theoretical value)
82.8%) mp118~120℃. Example 76 m-trifluoromethylphenyl-1-[2-
(bis-(2-chloroethyl)-amino)-2
-Oxo-tetrahydro-2H-1,3,2-
Oxazaphosphorin-4-yl-oxy]-urea 4.7 g (25 mmol) m-trifluoromethylphenyl isocyanate dissolved in 40 ml acetone
is added to 7.3 g (25 mmol) of aldophosphamide-oxime in 80 ml of acetone. The reaction mixture was stirred at 0 °C for 3 h and then in a condenser at -25 °C.
Leave it overnight. Then 150 ml of petroleum ether are added thereto and the mixture is left at −25° C. in a cooling box for another night. The crystals formed are separated, dried at 30°C and recrystallized from isopropanol.
Yield 9.2g (76.8% of theory), mp 91-93°C. Example 77 3-cyclohexyl-1-[2-(bis(2-
Chloroethyl)-amino)-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-oxy]-urea aldophosphamide-oxime 5 g (18 mmol) and cyclohexyl isocyanate 2.2 g
are separately dissolved in 10 ml of acetone and the solutions are mixed at 0°C. After standing for 2 hours, the crystals formed are filtered off with suction and recrystallized from acetone/ether. Yield 4.2g (56% of theoretical value),
mp113℃. Example 78 3-ethyl-1-[2-(bis-(2-chloroethyl)-amino)-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorine-
4-yl-oxy]-urea 5 g (18 mmol) of aldophosphamide-oxime and 1.2 g of ethyl isocyanate are each dissolved separately in 15 ml of acetone. The solutions are mixed at about 0°C. After standing for 5 hours, the crystals that have separated are filtered off with suction and washed with acetone/ether. Yield 3.5g (54% of theoretical value), m.
p.101℃. Example 79 2-(fluoren-2-yl)-1-[2-
(bis-(2-chloroethyl)-amino)-2
-Oxo-tetrahydro-2H-1,3,2-
Oxazaphosphorin-4-yl-oxy]-urea Fluorenyl-2- dissolved in 20 ml of acetone
2.1 g (10 mmol) of isocyanate in acetone
Aldophosphamide-oxime 2.9g in 30ml
(10 mmol) at 0°C. The separated crystals are separated the next day, dried under vacuum at 60°C and recrystallized from isopropanol/methanol. Yield 2.5g (50.1% of theory), mp114℃. Example 80 3-benzoyl-1-[2-(bis-(2-chloroethyl)-amino)-2-oxo-tetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-oxy]- Urea 2.9 g (20 mmol) of benzoyl isocyanate dissolved in 40 ml of acetone are added to 5.8 g (20 mmol) of aldophosphamide-oxime in 60 ml of acetone. The reaction mixture is stirred for 5 hours under a stream of nitrogen while cooling in an ice bath. The solid material that separates out is filtered off with suction, dried in a rotary evaporator at 30° C. and recrystallized from methanol. Yield 2.4
g (27.3% of theoretical value), mp124-125℃. Example 81 3-p-nitrophenyl-1-[2-(bis-
(2-chloroethyl)-amino)-2-oxotetrahydro-2H-1,3,2-oxazaphosphorin-4-yl-oxy]-urea 3.3 g p-nitrophenyl isocyanate dissolved in 40 ml acetone (20 mmol) in 5.8 g of aldophosphamide-oxime in 60 ml of acetone
(20 mmol). After standing for 2 hours, the solid material that separates out is filtered off with suction, dried in a rotary evaporator at 40° C. and recrystallized from DMF/ethanol. Yield 6.7g (73.5% of theoretical value), m.
p.117-118℃. Example 82 Tablets Coated with Gastric Juice-Resistant Coating 100 g of the compound of Example 5 are pressed through a sieve with 7.0 g of Aerosil (ie, finely divided amorphous silicic acid) and mixed thoroughly. Add to this mixture 76.0 g of Avicel PH105 (i.e. FMC's microcrystalline cellulose product), corn starch
Add 10g and 7.0g stearic acid. Mix this mixture until all ingredients are homogeneously dispersed. The mixture is compressed in the usual manner to produce tablet cores weighing 200 mg each containing 100 mg of active compound. Their tablet cores are coated with conventional gastric juice-resistant coatings such as suitable cellulose derivatives in organic solutions or aqueous dispersions (which may contain conventional plasticizers, dyes, sweeteners or antifoaming agents) or completely Cover with a synthetic coating. Example 83 Gelatin Capsules Coated with a Gastric Juice-Resistant Coating 250 g of the compound of Example 5 are pressed through a sieve with 7.5 g of Aerosil and mixed thoroughly. 40g lactose and magnesium stearate
Add 2.5 g to this mixture and mix it until the ingredients are homogeneously dispersed. This product is filled into gelatin capsules to produce capsules each containing 300 mg (dose) of active compound. The capsules are sealed and coated as described in Example 82.

Claims (1)

[Claims] 1. General formula {However, in the formula, Z is a group [formula] or a group -OR ₇ , X is oxygen or sulfur, and R ₁ , R ₂ and R ₃ are the same or different and are hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl, the radicals R ₄ are the same or different and represent hydrogen, methyl or ethyl;
R ₅ and R ₆ are the same or different, hydrogen, C _1-4
-Alkyl, hydroxy-C _1-4 - represents alkyl or phenyl, R ₇ is hydrogen, carbamoyl group,
-OR ₈ (In the formula, R ₈ is hydrogen, C _1-4 - alkyl,
phenyl or benzyl), straight-chain or branched C _1-18 -alkyl [possibly the same or different, hydroxy, halogen, -COOH,
-COOR ₉ , -CONH ₂ , -phenyl, benzyloxycarbonyl, -N(R ₉ ) ₂ , -N(R ₉ ) ₃ , -OR ₉ , -SR ₉ , -SO-R ₉ , -SO ₂ -R ₉ , _-SO3H or -PO( _CH3 ) ₂ (in the formula, _R9 represents methyl or ethyl) -
phenyl-C _1-4 -alkyl (optionally substituted with phenyl and/or
or substituted with one or two carboxy groups on the alkyl moiety), allyl,
C _3-8 - cycloalkyl, tetrahydrofuranyl,
Tetrahydropyranyl, phenyl (optionally 1
species or two of C _1-4 -alkyl, C _1-2 -alkoxy, nitro, halogen, trifluoromethyl, -
SO ₂ NH ₂ , substituted by carboxy, benzyloxycarbonyl and/or carbo-C _1-4 -alkoxy), benzyl, benzhydryl,
naphthyl, fluorenyl, pyridyl, thienyl,
benzoyl or C _1-4 -alkanoyl,
or R ₅ and R ₆ or R ₆ and R ₇ together with the atoms to which they are bonded optionally include an oxygen atom, a nitrogen atom substituted with C _1-4 -alkyl or -S-, -SO - or - form a saturated heterocycle containing a SO ₂ - group, or R ₆ and
R ₇ together with the adjacent nitrogen forms an aziridine ring optionally substituted with a cyano or carbamoyl group} 4-carbamoyloxy-oxazaphosphorine and its pharmaceutically useful salts. 2 Z is [Formula], X is oxygen, R ₅ and R ₆ are the same or different and are hydrogen, methyl or ethyl, and R ₇ is hydrogen, linear or branched C ₁ 4-carbamoyloxy-oxazaphosphorine according to claim 1, which is ₁₈ -alkyl, phenyl or benzyl. Claim 1 or 2, characterized in that all R ₄ groups in the formula 3 are hydrogen atoms.
4-Carbamoyloxy-oxazaphosphorine described in Section 1. 4 X is oxygen, R ₁ , R ₂ and R ₃ are the same or different and represent hydrogen or a 2-chloroethyl group, R ₄ , R ₅ and R ₆ represent hydrogen, and
R ₇ is hydrogen, benzyl, phenyl (in some cases 1
(substituted by one or two carboxy groups), C _1-4 -alkyl (optionally substituted by one carboxy group) or phenyl-
C _1-4 -alkyl (optionally 1 or 2 in the phenyl and/or alkyl part of the group)
4 of claim 1, characterized in that
-Carbamoyloxy-oxazaphosphorine. 5 formula (wherein R ₁ , R ₂ , R ₃ and R ₄ are as defined below and Y is hydrogen, methyl or ethyl) optionally heated in the presence of an inert solvent or While cooling and/
or in the presence of an acid catalyst (wherein Z, R ₅ and X are as shown below) {However, in the formula, Z is a group [formula] or a group -OR ₇ , X is oxygen or sulfur, and R ₁ , R ₂ and R ₃ are the same or different and are hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl, the radicals R ₄ are the same or different and represent hydrogen, methyl or ethyl;
R ₅ and R ₆ are the same or different, hydrogen, C _1-4
-Alkyl, hydroxy-C _1-4 - represents alkyl or phenyl, R ₇ is hydrogen, carbamoyl group,
-OR ₈ (In the formula, R ₈ is hydrogen, C _1-4 - alkyl,
phenyl or benzyl), straight-chain or branched C _1-18 -alkyl [possibly the same or different, hydroxy, halogen, -COOH,
-COOR ₉ , -CONH ₂ , -phenyl, benzyloxycarbonyl, -N(R ₉ ) ₂ , -N(R ₉ ) ₃ , -OR ₉ , -SR ₉ , -SO-R ₉ , -SO ₂ -R ₉ , _-SO3H or -PO( _CH3 ) ₂ (in the formula, _R9 represents methyl or ethyl) -
phenyl-C _1-4 -alkyl (optionally substituted with phenyl and/or
or substituted with one or two carboxy groups on the alkyl moiety), allyl,
C _3-8 - cycloalkyl, tetrahydrofuranyl,
Tetrahydropyranyl, phenyl (optionally 1
species or two of C _1-4 -alkyl, C _1-2 -alkoxy, nitro, halogen, trifluoromethyl, -
SO ₂ NH ₂ , substituted by carboxy, benzyloxycarbonyl and/or carbo-C _1-4 -alkoxy), benzyl, benzhydryl,
naphthyl, fluorenyl, pyridyl, thienyl,
benzoyl or C _1-4 -alkanoyl,
or R ₅ and R ₆ or R ₆ and R ₇ together with the atoms to which they are bonded optionally include an oxygen atom, a nitrogen atom substituted with C _1-4 -alkyl or -S-, -SO - or - form a saturated heterocycle containing a SO ₂ - group, or R ₆ and
R ₇ together with the adjacent nitrogen forms an aziridine ring optionally substituted with a cyano or carbamoyl group. 6 together with physiologically acceptable carriers and/or diluents of the general formula {However, in the formula, Z is a group [formula] or a group -OR ₇ , X is oxygen or sulfur, and R ₁ , R ₂ and R ₃ are the same or different and are hydrogen, methyl, ethyl, 2-chloroethyl or 2-methanesulfonyloxyethyl, the radicals R ₄ are the same or different and represent hydrogen, methyl or ethyl;
R ₅ and R ₆ are the same or different, hydrogen, C _1-4
-Alkyl, hydroxy-C _1-4 - represents alkyl or phenyl, R ₇ is hydrogen, carbamoyl group,
-OR ₈ (In the formula, R ₈ is hydrogen, C _1-4 - alkyl,
phenyl or benzyl), straight-chain or branched C _1-18 -alkyl [possibly the same or different, hydroxy, halogen, -COOH,
-COOR ₉ , -CONH ₂ , -phenyl, benzyloxycarbonyl, -N(R ₉ ) ₂ , -N(R ₉ ) ₃ , -OR ₉ , -SR ₉ , -SO-R ₉ , -SO ₂ -R ₉ , _-SO3H or -PO( _CH3 ) ₂ (in the formula, _R9 represents methyl or ethyl) -
phenyl-C _1-4 -alkyl (optionally substituted with phenyl and/or
or substituted with one or two carboxy groups on the alkyl moiety), allyl,
C _3-8 - cycloalkyl, tetrahydrofuranyl,
Tetrahydropyranyl, phenyl (optionally 1
species or two of C _1-4 -alkyl, C _1-2 -alkoxy, nitro, halogen, trifluoromethyl, -
SO ₂ NH ₂ , substituted by carboxy, benzyloxycarbonyl and/or carbo-C _1-4 -alkoxy), benzyl, benzhydryl,
naphthyl, fluorenyl, pyridyl, thienyl,
benzoyl or C _1-4 -alkanoyl,
or R ₅ and R ₆ or R ₆ and R ₇ together with the atoms to which they are bonded optionally include an oxygen atom, a nitrogen atom substituted with C _1-4 -alkyl or -S-, -SO - or - form a saturated heterocycle containing a SO ₂ - group, or R ₆ and
R ₇ together with the adjacent nitrogen forms an aziridine ring optionally substituted with a cyano or carbamoyl group} Cell proliferation inhibitor containing 4-carbamoyloxy-oxazaphosphorine as an active ingredient .