IL28263A - Cyclic phosphoric acid derivatives and their production - Google Patents

Description

28263/2 nsoinn *tv mvnn ηΐ'^ρ».. ma ram New cyclic phosphoric acid derivatives and their production ASTA-WER A TIENGESELLSCHAFT CH ISCHE! FABRIK She present invention consists in new cyclic phosphoric acid derivatives of formula wherein R^ S a lower alfcyl group having from 1 to 4 carbon atoms an being substituted with one or more halogen atoms, the Z are independently hydrogen or a lower allcyl group having from 1 to 4 carbon atoms, is 2 or 5* and Xg is an ethylene imino grou o a group of the formula 28263/2 - 8 - compound of formula is reacted with a compound of formula wherein is halogen, preferably chlorine, R.^, 2, Z and m have the same meaning as in formula I and Rc has the same meaning as in formula XI in the presence of an acid binding agent; at an elevated temperature, preferabl in the range of about 100^C. to about 250°C.

Preferably, the reactions A, B,and C are' carried out in an inert organic solvent such as a halogenated lower ali-phatic hydrocarbon such as chloroform or methylene dichlorid l - : ■·! ' '· .■ or in an aromatic hydrocarbon such as benzene or toluene, or in an ether such as diethyl ether .or dioxane. The acid binding agent should be. present in an amount corresponding .· to at least two mol equivalents: in the metho embodiment A and in an amount corresponding to at least one mol equiva-lent in the method embodiment B in order to preferably absorb the total amount of. acid HX^ produced ^during the reactions. Many basic compounds are known tosttie expert useful as acid binding agent such as alkali metal carbonates and i bicarbonates and particularly tertiary amines such as triethyl amine or pyridine. The reactions A, r-B, arid C may be carried out at room temperature or at an elevated: . temperature such as a temperature elevated t6 the boiling point of the solvent usedv Halogenating agents for . . exchanging aliphatically. 'bound hydroxy groups' to halogen atoms are known to the expert, too. Such agents' are for instance the phosphorous trihalogenides such . ats iphosphorus trichloride PCl^ and phos.phoDrus tribromide PBr^1, the phosphoric acid halbgenides such' as phosphorous- penta- chloride PCl^, phosphorous : oxychloride POCl-j and ! phosphorous oxybromide, the halogenides of sulphurous and sulphuric Determination of CD 50 (4) with a Yoshida-Ascites- Sarkom of rats with a total dose 'D. subdivided ..into- 4 subsequent days. Start up of therapy at the -day of sarkom implantation; Test period: 90 days. ■ Determination of LD 50 (1) with a single administration- to. rats of the same race; Test period: 1 4 days.

The compound according. to Example 1 is characterized by a very good · chemotherapeutic effectiveness -to the mouse tumours Sarkoma 37 and Ehrlich-Carcinoma known to be ' resistant to chemotherapy; the known comparative test compound A is not effective or exerts · effectiveness, only with subtoxic doses. The compound of Example 4 is characterized over the known test compound by that the accumulation of the toxic effectiveness is smaller in the lethality test..

Thus, the LD 50 (4) of compound A is 140 mg.-/kg. and thus lower, than the LD 50 (1) while. the LD 50 (4) of .the com- , pound according to Example 4 amounts ■ o 190 mg>/kg. and thus is significantly higher than the LD 50 (1). This .: lower toxic accumulation is of importance in the ' treatment over prolonged periods of .time.

■Furthermore, the compounds of Formula V is characterized by a high solubility .in water and furthermore represents, a new type of cytostatics wherein the 2-chloroeth 1 groups are not bound to one and the same nitrogen atom as in the ■ nitrogen mustard compound; on the contrary, the tv/o 2-chloro- ethyl groups are substituted at two different amide nitrogen atoms .

Finally, in comparison to the known cytostatic N,N-bis-B-chloroethylamino- 1 , O-propylene-phosphoric acid ester diamide- (test compound A) which is well accepted in the medical art under the generic name Cyclophosphamide , the compounds of the present invention are characterized by an improved stability in aqueous solutions as follows from Table II. When allowing compounds of the. present invention to stand in a bicarbonate buffer solution (pH 7.5) for -10 days at 37«5°C, chlorine ions are split off in an amount of only about 1/6 of the amount split off . with the known test compound A.

Table II The following Examples serve to further illustrate^ the ·;. present invention without however, limiting the same thereto.

Example 1 Ν,Ν,Ν ' -Tris-( 2-chloroethyl) -N ' , O-propylene phosphoric ■· acid ester diamide 2.59 g. (Ί mol) of N,N-bis-( 2-chloroethyl) phosphoric acid amide dichloride, 209 g. (1.2 mol) of N-( 2-chloroethyl) - N-( 3-hydroxypropyl) -amine hydrochloride (crude), 1000 cc. of methylene dichloride and 344 g. ( 3 · 4' mol).. of triethyl- amine . ■ · ' N,N-bis-( 2-chloroethyl) -phosphoric acid amide dichloride is dissolved in the methylene dichloride. N-( 2-chloroethyl) N-( 3-hydroxypropyl) -amine hydrochloride is suspended in this solution and, triethylamine is added .thereto dropwise with stirring. The temperature of the solution rises, to "boiling. After the termination of the addition, the mixture is heated to boiling for another 6 hours. There-, after, the reaction mixture is cooled down and allowed to •stand' over night at about 0°C. The. precipitated triethyl- amine hydrochloride, is filtered off with suction. The resulting solution is evaporated, the residue (about .370 g. is triturated- ith about 3.2 1. of ether and is heated to boiling for a short period 'of time. The ethereal solution is decanted from the insolubles (about 90 g.). The solution is rendered to pH 6.5 to.7 by the addition -of ethereal hydrochloric acid and then is filtered over charcoal and thereafter; is evaporated . During evaporation, the temperature should not rise above 40°C. The residue is dissolved in ether in an amount corresponding to half of its weight (240 g. of residue dissolved in 120 cc. of ether) , the . ethereal solution is cooled to -5°C« and. is inocculated .

After standing for 25 hours, 140 g. "have bee separated . by crystallization. After separation by filtration with suction, the mother liquor is diluted with ether to: 5 times its volume, the solution is filtered over charcoal, is · again evaporated and the residue' is . again dissolved in a volume corresponding to .half of the weight of the residue.

Another cooling to -5°C. and inocculation produces further termination of the addition, the reactio mixture is stirred at 50°C . for another three, hours ." Thereafter , the mixture is cooled and precipitated trieth lamirie hydrochloride is separated by filtration with suction.

The mother liquor is evaporated in a vacuum and the resi-.. due is dissolved in 500 cc- of ether. This solution. is washed 3 times with water,- then washed with dilute soda lye and finally washed another time with water. Thereafter' it is dried over anhydrous sodium sulphate and evaporated in a vacuum to about 1 30 g. This solution is cooled down to -5°C. and inocculated, thus causing- crystallization and separation of 28 g. of the desired product. After ~~ separation thereof the mother liquor is diluted with' ether to about five times the initial volume. It is then filtered over charcoal and evaporated. The residue is dissolved in suc a volume of ether which corresponds to half of the'weight of the residue. The solution is. again, cooled down-to -5°C . Upon inocculation, another 1 0 g. of the desired compound crystallizes .. Total yield: 38 g. (47 of .the theoretical) .

Pp. : 50 to 51.°C.

Example 3 .. Ν,Ν,Ν ' -Tris-( 2-chloroethyl) -N ' , 0-propylene phosphoric acid ester diamide, 42.6 g. ( 0 . 30 mol) of ,N-bis-( 2-chloroethyl) -amine , N-chloroethyl-N, 0-propylene phosphoric acid ester amide. . monochloride and 65 cc« -^of methylene dichloride. : ; . 42 . 6 g'. of N,N-"bis-( 2-chloroethyl)-amine are dissolved in' 1 50 cc. of methylene dichloride. A solution of 32 . 7 g. of ' N-( 2-chloroethyl)-N, 0-propylene phosphoric acid ester amide monochloride in 65 cc. of methylene dichloride are added' thereto with stirring. ' The mixture is heated to boiling for 1 hour and thereafter evaporated in a' vacuum. The residue is triturated with 350 cc. of anhydrous, ether ' and precipitated Ή ,N-bis-( 2-chloroethyl) -amine hydrochloride is separated by filtration with suction.. The. ethereal solution is washed, with dilute hydrochloric acid and /thereafter with a solution of sodium. bicarbonate. After drying over ■ anhydrous sodium sulfate, 'the ether is distilled off and · the oily residue is carefully dried, dissolved in ten times its volume of anhydrous ether, filtered over activa- : ted charcoal and evaporated again. The residue is dissolved in 40 cc. of anhydrous ether. After cooling at - 5 °C , • in '■ ' '· ■·· crystallization is initiated by peculation. After, standing. " 'for 24 hours, the precipitated crystals are filtered off' with suction and dried in a. vacuum. Yield: 24 · 3 g* ( 50 . 1 of the theoretical).

Pp. :.51 to 52 °0 . ' - -g - ■ Example N,N'-Bis-(2-chloroethyl)-N' ,0-propylene phosphoric acid ester diamide 127.6 g. (1.1.mol) of N-(2-chloroethyl) -amine hydrochloride- are suspended in a solution of 218 g. (1 mol) of N-(2- chloroethyl) -N , O-propylene phosphoric acid ester amide monochloride in 600 cc. of methylene dichloride, and 212 g. of triethylamine are added thereto dropwise with stirring. ' The reaction mixture is, heated to boiling by the reaction ■ heat. After termination of the addition, the reaction mixture is heated to boiling for another 2 hours . '. Thereafter, it is cooled to room temperature and the precipitated triethylamine hydrochloride' is separated, by filtration with suction. The filtrate is extracted with about 60' cc. of > dilute hydrochloric acid (pH 3)» then twice with .about 60 cc. of water, .thereafter with about 60 cc. of dilute soda lye and finally twice with about 60 cc. of. water..

After drying over, anhydrous .sodium sulfate, methylene dichloride is distilled off under normal pressure .' The oily residue is dried in a vacuum and thereafter extracted in a perforator with 500 cc. of anhydrous ether. The oily extract crystallizes upon inocculation. and standin in an ice box. After standing for several hours, the precipitate is filtered off, washed with a small amount of cold ether and dried in a vacuum at room temperature. Yield: 185 g. (71 c/ of the theoretical) .

Pp. : 39 to 41°C..

Example 5 ■" '.'■» · ' ' ·" ' N-Methyl-N ,Ν ' -bis-( 2-chloroethyl) -H ' , O-propylene. phosphoric acid, ester diamide 90.8 g. of trieth lamine are added to a suspension of ■52.2 g. of 2-chloroethyl-3-hydroxypropyl-amine hydrochloride in 2.10 cc. of methylene dichloride. A solution of 52.5 g. of N-methyl-:tT-( 2-chloroethyl) -phosphoric .acid. ' amide dichloride in 52.5 cc. .of ' methylene dichloride is added, thereto with stirring. During t e addition,- the reaction mixture is heated to boiling by the reaction heat.-After- termination of the ^addition,, the reaction mixture is', heated to boiling for another 5 hours.- After cooling to room temperature., the precipitated, triethylamine hydrochloride is filtered off' with suction. The filtrate is washed subsequently with dilute hydrochloric acid, water, 2 N-soda lye and then twice with water. After 'drying over, anhydrous sodium sulfate, the solution is .evaporated in ·-.' a vacuum and the oily residue is dissolved, in twelve times its volume of ether. The resulting' solution is filtered over activated, charcoal and..-is again evaporated. The colourless oil is dried in a vacuum. Yield: 52 g.-. (75.51° of the theoretical).

Example 6 ■■ . Έ ,N-Ethylene-I\i '-( 2-chloroethyl) -N 1 , O-propylene .phosphoric acid, ester diamide .

A ' solution of 0.72 g. of ethylene imine and 11.1 g. -of triethylamine in 30 cc. of ether are added with stirring to a solution of 21 . 8 g. of N-( 2-chloroethyl) -N , O-propylene. phosphoric acid ester amide .monochloride' in 1 32 cc. of . anhydrous ether. The temperature of the reaction mixture 'rises and triethylamine hydrochloride is precipitated.

After termination of the addition, the reaction mixture is, stirred at room temperature for another 2 hours.

Thereafter, precipitated, triethylamine hydrochloride is filtered off with suction, the filtrate is filtered over activated charcoal and the solvent is distilled off. The .. ' oily residue is carefully dried in a vacuum and' is then ■ dissolved in, ten times its. volume of. anhydrous- ether . The solution is filtered over activated charcoal onc more and . is. again evaporated. The oily residue is carefully dried in' a vacuum. Yield: 1 5 g« ' ( 67 i° of, the. theoretical).

Example 7 ', - c .

N ,N-Bis-( 2-chloroeth l) -N ' -( 3-chloropropyl) j-"' , 0-propylene phosphoric acid, ester diamide . 1 31.9 g. ( 1 /1 0 moi') of N,N-bis-( 2-chloroethyl) 3- . ■ · . ■ :<■ hy'droxypropyl)-W , 0-propylene phosphoric acid, ester- '. diamide are dissolved. ih-80¾ cc. of chloroform and a solu- tion of 1 4 cc. ( 2/1 0 mol) of ' thionylchlorid¾ in 20 cc of chloroform are added thereto with stirring.'-. There occurs a weakly exothermic reaction. After termination of the addition, nitrogen is huhhled through the reaction mixture wh'ile heating the mixture to a temperature of f„38 to: 0°C .

, : ■ . . : -. Formation of HC1 and -SOg is . terminated, after 3 hours .

After coolin to room temperature,- the solution is shaken with 0.1 IT hydrochloric acid until a remaining acid reaction, of the mixture. Thereafter, the mixture is neutralized by washing with dilute sodium bicarbonate solution and the neutralized chloroform solution is dried over anhydrous sodium sulfate. After separation of sodium sulfate, chloroform is-.distilled : off in a vacuum. ;and the oily residue is carefully dried. Yield: 16 g... (47.4. f of the theoretical).

Example 8 N,N-Bis-( 2-chloroeth l) -N ' -( 3-chloropropy1) -N ' , 0-propylene phosphoric acid ester diamide .9 g' of N,N-bis-(2-chloroethyl)-phosphoric acid amide dichloride (0.1 mol) , 18.8 g.' of N-( 3-chlor'opropyl) -N-( 3-hydroxypropyl) -amine hydrochloride (0.1 mol), 30.3 g-triethylamine (0.3 mol), and 100 cc. of. dioxane.: The above starting materials are heated at,50°C. with stirring for 3 hours.' After cooling to room temperature , the precipitated, tr ethylamine hydrochloride' is filtered, off with suction and the .solution is evaporated in a vacuum. The- oily residue is dissolved in 250 cc. of ether r and the solution is shaken with 0.1 IT hydrochloric acid, until the solution remains acid. Thereafter, the solution is neutralized by v/ashing with a dilute sodium bicarbonate ' Example 11 >N'-Bis-(2-chloroethyl)-N'<0-¾utylene- ,3 phosphoric acid ester diamide . .

A solution' of 29 g. of triethylamine and .25 cc. of methylene chloride is added dropwise with stirring to a solution of 32 g. of N~(2-chloroethyi)-N,0-butylene- ,3 phosphoric acid ester amide monochloride (i.e. 2-chloro-.3- ( 1 -chloroethyl)-4~ rnethyl-tetrahydro-2H- , 3 » 2-oxaza-phosphorine-2-oxide) and 17.7 g. of N-(2-chloroethyl) amine hydrochloride and 25 cc. of methylene chloride. The mixture is heated' to boiling' for two; ho and cooled to room temperature therafter. The., precipitated triethylamine hydrochloride · is filtered off with suction.

The remaining solution is shaken consecutively with 15 cc. of dilute hydrochloric acid, 1o cc. of water,. 1o cc. of an · . ' aqueous sodium bicarbonate solution and finally twice with o cc'. of water.

The methylene chloride solution is dried over anhydrous sodium sulfate and evaporated in a vacuum. The oily residue is disaolv in four times the amount of anhydrous ether and the solution- is filtered over activated charcoal and evaporated in a vacuum. Yield in N, N' -bis'~( 2-chloroethyl.)- ' , 0-butylene-1 , phosphoric acid ester diamide o -3- (2-chloro- ethyl)- -methyl-tetrahydro-2H-1 , 3» 2-oxaza-phosphorine~2-oxidc : 32.3 g. (85; % of the theoretical) . ' *' ■: ■ n D" : .'l,5o67. ! ·■ - '■.■■· ;·'.. ,' 'n· '. .■·· Example 1 , N1 -Bis- ( 2-chloroethyl) -N' 0-2 ; 2-dimethyl-pronylene- , 3 phosphoric acid, ester diamide 2o.2 g.- of 2, 2-diraethyl- -(-2-chloroethyl)-amino-propanol-( ,3)-hydrochloride (l/1o.mol) are suspended i 16ο cc. of anhydrous chloroform. 31 g. of triethylamine (3/1o mol) are added thereto stepwise with shaking and cooling with ice.

The resulting clear solution is added to a solution of 19.6 g. of N-(2-chloroethyl) phosphoric acid amide dichloride ( 1/1 o mol) in 2o cc. of anhydrous chloroform over a period of one hour dropwise with- stirring at a temperature of the reaction mixture of· 2o to 25°C. Stirring is continued for two hours at 4o°C. Thereafter, the solvent is evaporated in a water Jet vacuum and the semi-solid residue is mixed -with 1oo cc. of ether. The oily, component of- the residue thus . is dissolved and the remaining crystalline part consists of ■ triethylamine hydrochloride. The crystals are filtered off )■ with suction and the remaining solution, i , evaporated. The · remaining' oil is purified by extractio with' 3:1 ether.:petrol ether in .a perforator. The clear colorless oil is' soluable in alcohol, chloroform, dioxane and acetone. .' Yield in N,N,-bis-(2-chloroethyl)-N' , 0-2, 2-dimethyl-propylene- 1,3 phosphoric acid ester diamide or 2-(2-chloroethyl amino ) ~3- '(•2-chloroethyl )-5-dimethyl-2H-tetrahydro- , 3, 2-oxaza-phos'phorine 2-bxide :.'. '■ " · . ' .· . .· . s . ' · ' .;· . · ■ ■'■ ■·'■"' 12,9 g. '(4 .6% of the theoretical) . v ,: ■■■ 22 ' ■ '·' ·' .' ' .·-' ■' n y. :..1.5p 2 -> ' '.' ' ' ■. . ' ■ : ; j The. oil .may be solidified, by dissolving in chloroform,- washing the solution consecutively with i-/ o hydrochloric acid, 50 aqueous sodium bicarbonate solution and water, drying the solution over anhydrous calcium chloride and evaporation of the solvent. The residue is dissolved in a small amount of chloroform and the solution is passed through a -column of 1"io g. of silica gel. The compound then is eluated by means. of Soo cc. of ' chloroform.

Th.e solvent is evaporated and the remaining oil is di-ssolved in a small amount of ether 'and the solution is cooled in an icebox. · Crystallization is initiated by- friction with a. las rod. ' Example *I5 N , N, M ' -Tris- ( 2-chloro ethyl ) - ' , 0-2 , 2-dimethyl-propylene- 1 , 3 phosphoric 'acid ester diamide 31 g. of triethylaraine ( 3/1O mol) are added in 'small amounts with stirring and cooling to a suspension of 2o . 2 g. of N- ( 2-chloroethyl) -2 , 2-dimethyl-amino-propanol- ( Ί> 3) hydrochloride ( 1 / 1O mol) in 15o cc. of anhydrous chloroform. The resulting clear solution is- added to a solution of 25. 9 g.. of N,N-Bis- ( 2-chloroethyl) phosphoric acid amide dichloride ( /1 o mol) in 13o cc. of anhydrous chloroform over a period of 15 minutes dropwise at 25°C.

Stirring at 4o°C. is continued for two hours., In. order to completely separate the .resulting triethylamine hydrochloride, 5o cc. of ether are. added to the mixture which . then ;is .allowed to stand in ah icebox for 2o hours.

The precipitated. hydrochloride is separated by filtration and the resulting solution is evaporated in a water jet vacuum. The oily residue is extracted in a .perforator with 1.: 2 e,ther:petrol ether. After evaporation of the eluant, the oily residue is dissolved in methylene chloride. The solution is washed consecutively v/ith 1 /2o N hydrochlorid acid, '5% aqueous sodium bicarbonate solution and water. · "· The remaining solution is dried over anhydrous ' sodium sulfate After removing of the solvent in a water jet vacuum, ' the oil is dried in a high vacuum. The resulting clear colorless oil is solua'ble in alcohol, chloroform, acetone and dioxane. Yield in N, N, N.' -tris- ( 2-chloroethyl)-N' , 0-2 , 2-dimethyl- " · propylene- 1', 3 phosphoric acid ester diamide or 2 -/Ny -bis- "' ■.. · i' I

Claims (1)

1. 28263/3 28263/2» - 35 - 16. Processes for the preparation of cyclic phosphoric acid derivatives according to Claim 1, substantially as described herein with reference" to the Bxamples. PC.BH