Patent Publication Number: US-4546181-A

Title: Platinum chelates of 3-amino-substituted-[1,2,4]triazolo[1,5-a]-pyrimidin-7(3H)-ones

Description:
SUMMARY OF THE INVENTION 
     This invention is concerned with new compounds which may be represented by the following formula I: ##STR1## wherein R 1  is selected from the group consisting of hydrogen and formyl; R 2  is selected from the group consisting of hydrogen, alkyl(C 1  -C 3 ) and phenyl; R 3  is selected from the group consisting of hydrogen and alkyl(C 1  -C 3 ); R 4  is selected from the group consisting of alkyl(C 1  -c 3 ) and phenyl and L and L&#39; are the same and are selected from the group consisting of halide, nitrate, sulfate and a monobasic organic acid, such as glucuronic acid, or L and L&#39; taken together may be a dibasic organic acid such as malonic asid, oxalic acid, methyl malonic acid, succinic acid, tartonic acid, or cyclobutane dicarboxylic acid. 
     In addition this invention is concerned with a class of intermediates used to prepare the compounds of formula I, which intermediates may be represented by the following formula II: ##STR2## wherein R 1 , R 2 , R 3  and R 4  are as described for Formula I. 
     DESCRIPTION OF THE INVENTION 
     The compounds of this invention may be prepared according to the following reaction sequence: ##STR3## 
     In accordance with the above reaction scheme, a substituted 3-amino-2-hydrazino-4-(3H)-pyrimidinone (I) is reacted with triethyl ortho derivative (2) in n-butanol at reflux 30-60 minutes, giving the 3-amino-substituted-[1,2,4]triazolo[1,5-a]pyrimidin-7(3H-one intermediate (3) where R 2 , R 3 , and R 4  are as described above. 
     Alternatively (1) is reacted with formic acid at reflux for 12-48 hours, giving the intermediate (4). 
     Intermediates (3) or (4) are then reacted with potassium tetrachloroplatinate in aqueous solution for 12-48 hours, giving the products (5). 
     To prepare compounds of formula I where L and L&#39; are nitro, compounds of formula I where L and L&#39; are chloro are dissolved in water and reacted with 2 equimolar amounts of silver nitrate for several hours at room temperature, then filtered and the filtrate evaporated to give the dinitrate derivatives. 
     To obtain the organic acid derivatives the dinitrate compound is reacted with one equimolar amount of the potassium salt of a dibasic acid or 2 equimolar amounts of the potassium salt of a monobasic acid. 
     The novel chelated compounds of the present invention possess the property of inhibiting the growth of transplanted mouse tumors as established by the following test. 
     Lymphocytic leukemia P388 test 
     The animals used were BDF/1 mice all of one sex, weighing a minimum of 18 g and all within a 3 g weight range. There were 5 to 6 animals per test group. The tumor transplant was by intraperitoneal injection of 0.5 ml of dilute ascitic fluid containing 10 6  cells of lymphocytic leukemia P388. The test compounds were administered intraperitoneally on days 1,5 and 9, relative to tumor inoculation, at various doses. The animals were weighed and survivors recorded on a regular basis for 30 days. The median survival time and the ratio of survival time for treated (T)/control (C) animals were calculated. The positive control compound was either 5-fluorouracil, 1,4-dihydroxy-5,8-bis[[2-(2-hydroxyethylamino)ethyl]-amino]anthraquinone dihydrochloride or Cisplatin. The results of this test on representative compounds of this invention appear in Table I. 
     
                       TABLE I                                                     
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                           Median                                         
                           Sur-                                           
                  Dose     vival   T/C × 100                        
Compound          (mg/kg)  (Days)  (%)                                    
______________________________________                                    
3-amino-5-methyl[1,2,4]-                                                  
                  200      15      127                                    
triazolo[1,5-a]pyrimidin-                                                 
                  50       15      127                                    
7(3H)-one, compound with                                                  
                  12       12.5    106                                    
platinum chloride (1:1)                                                   
Control           --       11.8    --                                     
5-Fluorouracil    50       20.5    174                                    
                  12       15      127                                    
                   3       13.5    114                                    
3-amino-2,5-dimethyl[1,2,4]-                                              
                  200      15      127                                    
triazolo[1,5-a]pyrimidin-                                                 
                  50       12.5    106                                    
7(3H)-one compound with                                                   
                  12       12      102                                    
platinum chloride (1:1)                                                   
Control           --       11.8    --                                     
5-Fluorouracil    50       20.5    174                                    
                  12       15      127                                    
                   3       13.5    114                                    
3-amino-5-phenyl-[1,2,4]-                                                 
                  200      11.5    115                                    
triazolo[1,5-a]pyrimidin-                                                 
                  50       10.5    105                                    
7-(3H)-one, compound with                                                 
                  12       11.5    115                                    
platinum chloride (1:1)                                                   
                   3       10.5    105                                    
Control           --       10      --                                     
1,4-dihydroxy-5,8-bis[[2-(2-                                              
                  1.6      &gt;30     &gt;300                                   
hydroxyethylamino)ethyl]-                                                 
                  0.4      19      190                                    
amino]anthraquinone dihydro-                                              
                  0.1      14.5    145                                    
chloride            0.025  16      160                                    
N(5,6-dimethyl-7-oxo[1,2,4]-                                              
                  200      18.5    185                                    
triazolo[1,5-a]pyrimidin-                                                 
                  50       13      130                                    
3(7H)-yl)formamide, compound                                              
                  12       13      130                                    
with platinum chloride (1:1)                                              
Control           --       10      --                                     
Cisplatin         0.8      23.5    235                                    
                  0.2      21      210                                    
3-amino-5-methyl-2-phenyl-                                                
                  200      15      150                                    
[1,2,4]triazolo[1,5-a]-                                                   
                  50       12      120                                    
pyrimidin-7(3H)-one, compound                                             
                  12       11      110                                    
with platinum chloride (1:1)                                              
Control           --       10      --                                     
Cisplatin         0.8      23.5    235                                    
                  0.2      21      210                                    
N--(5-methyl-7-oxo[1,2,4]-                                                
                  200      15      150                                    
triazolo[1,5-a]pyrimidin-                                                 
                  50       14.5    145                                    
3(7H)-yl)formamide, compound                                              
                  12       16      160                                    
with platinum chloride (1:1)                                              
                   3       11      110                                    
Control           --       10      --                                     
Cisplatin         0.8      23.5    235                                    
                  0.2      21      210                                    
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     This aspect of the invention includes the novel compositions of matter and the method of inducing the regression and/or palliation of leukemia and related cancers in mammals when administered in amounts ranging from about one mg to about 1.2 gm per square meter of body surface area per day. The interrelationship of dosages for animals of various sizes and species and humans (based on mg/m 2  of surface area) is described by Freireich, E. J., et al., Quantitative Comparison of Toxicity of Anticancer Agents in Mouse, Rat, Hamster, Dog, Monkey and Man. Cancer Chemother. Rep., 50, No. 4, 219-244, May 1966. A preferred dosage regimen for optimum results would be from about 3 mg/m 2  /day to about 200 mg/m 2  /day, and such dosage units are employed that a total of from about 5 mg to about 360 mg of the active compound for a subject of about 70 kg of body weight are administered in a 24 hour period. This dosage regimen may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. The active compound may be administered by the intravenous, intramuscular, or subcutaneous routes. 
     The active compounds may be administered parenterally or intraperitoneally. Solutions or dispersions of the active compound can be prepared in water suitably mixed with a surfactant such as hydroxypropyl cellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prvent the growth of microorganisms. 
     The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent of dispersion medium containing, for example, water, ethanol, polyol, (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases it will be preferable to include isotonic agents, for example sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example aluminum monostearate and gelatin. Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredient into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze-drying technique which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. 
     As used herein, &#34;pharmaceutically acceptable carrier&#34; includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatable with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. 
     It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the novel dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active material for the treatment of disease in living subjects having a diseased condition in which bodily health is impaired as herein disclosed in detail. 
     The principal active ingredient is compounded for convenient and effective administration in effective amounts with a suitable pharmaceutically acceptable carrier in dosage unit form as hereinbefore disclosed. A unit dosage form can, for example, contain the principal active compound in amounts ranging from about 2 mg, to about 2 g, with from about 5 to about 360 mg being preferred. Expressed in proportions, the active compound is generally present in from about 2 to about 100 mg/ml of carrier. In the case of compositions containing supplementary active ingredients, the dosages are determined by reference to the usual dose and manner of administration of the said ingredients. 
     Regression and palliation of cancers are attained, for example, using intraperitoneal administration. A single intravenous dosage or repeated daily dosages can be administered. Daily dosages up to about 5 or 10 days are often sufficient. It is also possible to dispense one daily dosage or one dose on alternate or less frequent days. As can be seen from the dosage regimens, the amount of principal active ingredient administered is a sufficient amount to aid regression and palliation of the leukemia or the like, in the absence of excessive deleterious side effects of a cytotoxic nature to the hosts harboring the cancer. As used herein, cancer disease means blood malignancies such as leukemia, as well as other solid and non-solid malignancies such as the melanocarcinomas, lung carcinomas, and mammary tumors. By regression and palliation is meant arresting or retarding the growth of the tumor or other manifestation of the disease compared to the course of the disease in the absence of treatment. 
     This invention will be described in greater detail in conjunction with the following specific examples. 
    
    
     EXAMPLE 1 
     3-Amino-5-methyl[1,2,4]triazolo[1,5-a]pyrimidin-7(3H)-one 
     A mixture of 26.44 g of diaminoguanidine hydrochloride, 10.8 g of sodium methoxide and 200 ml of ethanol was stirred for 30 minutes and then filtered. A 13 g portion of ethyl acetoacetate was added to the filtrate. The crystals were collected and washed with cold ethanol, giving 7.62 g of 3-amino-2-hydrazino-6-methyl-4(3H)-pyrimidinone. 
     A 2.4 g portion of 3-amino-2-hydrazino-6-methyl-4(3H)-pyrimidinone was dissolved in 100 ml of n-butanol by heating to the boiling point. A 20 ml portion of triethyl orthoformate was added and the solution was refluxed for 45 minutes. The solid was collected while the solution was hot, washed with ether and dried, giving 1.45 g of the desired intermediate, mp 202°-204°. 
     EXAMPLE 2 
     3-Amino-2,5-dimethyl[1,2,4]triazolo[1,5-a]pyrimidin-7(3H)-one 
     A 3.0 g portion of 3-amino-2-hydrazino-6-methyl-4(3H)-pyrimidinone, 20 ml of triethyl orthoacetate and 100 ml of n-butanol was reacted as described in Example 1, giving 2.36 g of the desired intermediate, mp 287°-289° C. 
     EXAMPLE 3 
     N-(5,6-Dimethyl-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-3(7H)-yl)formamide 
     A mixture of 13.22 g of diaminoguanidine hydrochloride, 5.4 g of sodium methoxide, 7.2 g of ethyl-2-methyl acetoacetate and 100 ml of ethanol was reacted as described in Example 1, giving 4.5 g of 3-amino-2-hydrazino-5,6-dimethyl-4(3H)-pyrimidinone. 
     A 2.0 g portion of the above compound and 20 ml of formic acid was refluxed overnight and then evaporated to dryness. The residue was crystallized from ethanol, giving 1.82 g of the desired intermediate, mp 232° C. (dec). 
     EXAMPLE 4 
     3-Amino-5-methyl-2-phenyl[1,2,4]triazolo[1,5-a]pyrimidin-7(3H)-one 
     A mixture of 2.0 g of 3-amino-2-hydrazino-6-methyl-4(3H)-pyrimidinone, 10 ml of triethyl orthobenzoate and 20 ml of n-butanol was reacted as described in Example 1, giving 1.27 g of the desired intermediate, mp 195° C. (dec.). 
     EXAMPLE 5 
     N-(5-Methyl-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-3(7H)-yl)formamide 
     A mixture of 1.0 g of 3-amino-2-hydrazino-6-methyl-4(3H)-pyrimidinone and 10 ml of formic acid was refluxed overnight, then cooled to room temperature and evaporated to dryness. The residue was slurried in methanol and then diluted with ether. The solid was collected and dried, giving 1.2 g of the desired intermediate, mp 218° C. (dec.). 
     EXAMPLE 6 
     3-Amino-5-phenyl[1,2,4]triazolo[1,5-a]pyrimidin-7(3H)-one 
     A suspension of 13.22 g of N, N&#39;-diaminoguanidine hydrochloride and 5.4 g of sodium methoxide in 150 ml of ethanol was stirred for one hour and then filtered. To the filtrate was added 9.6 g of ethyl benzoylacetate and this mixture was stirred one hour. The solid was collected, giving 4.8 g of 3-amino-2-hydrazino-6-phenyl-4(3H)-pyrimidinone. 
     A solution of 2.77 g of the above compound in 20 ml of triethyl orthoformate and 70 ml of n-butanol was refluxed for 1.5 hours, then cooled to room temperature and the solid was collected, washed with ether and dried, giving 1.39 g of the desired intermediate, mp 260°-266° C. 
     EXAMPLE 7 
     3-Amino-5-methyl[1,2,4]triazolo[1,5-a]pyrimidin-7(3H)-one, compound with platinum chloride (1:1) 
     A solution of 495 mg of 3-amino-5-methyl[1,2,4]triazolo[1,5-a]pyrimidin-7(3H)-one, 1.245 g of potassium tetrachloroplatinate and 30 ml of water was stirred overnight. The solid was collected, washed with water and dried, giving 1.08 g of the desired product, mp&gt;300° C. 
     Following the procedure of Example 7 and using the intermediates of Examples 2-6, the products of Examples 8-12, listed in Table II were obtained. 
     
                                           TABLE II                                
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     Intermediate                                                         
Example                                                                   
     of Example                                                           
            Product                MP °C.                          
__________________________________________________________________________
 8   2      3-amino-2,5-dimethyl[1,2,4]triazolo-                          
                                   &gt;300                                   
            [1,5- -a]pyrimidin-7(3 .sub.--H)--one, compound with          
            platinum chloride (1:1)                                       
 9   3       .sub.--N--(5,6-dimethyl-7-oxo[1,2,4]triazolo[1,5- -a]-       
                                   &gt;295                                   
            pyrimidin-3(7 .sub.--H)--yl)formamide, compound with          
            platinum chloride (1:1)                                       
10   4      3-amino-5-methyl-2-phenyl[1,2,4]triazolo-                     
                                   &gt;290                                   
            [1,5- -a]pyrimidin-7(3 .sub.--H)--one, compound with          
            platinum chloride (1:1)                                       
11   5       .sub.--N--(5-methyl-7-oxo[1,2,4]triazolo[1,5- -a]-           
                                   &gt;280                                   
            pyrimidin-3(7 .sub.--H)--yl)formamide, compound with          
            platinum chloride (1:1)                                       
12   6      3-amino-5-phenyl[1,2,4]triazolo[1,5- -a]-                     
                                   &gt;300                                   
            pyrimidin-7(3 .sub.--H)--one, compound with platinum          
            chloride (1:1)                                                
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     EXAMPLE 13 
     Preparation of Dinitrate Derivatives 
     To a suspension of 1 m mole of a dichloride derivative, selected from those of Examples 7-12, in 50 ml of water is added 2 m moles of silver nitrate. The mixture is stirred at room temperature for 3 hours and then filtered. The filtrate is evaporated to dryness, giving the desired dinitrate derivatives of formula I, where L and L&#39; are NO 3 . 
     To obtain the organic acid derivatives the dinitrate derivative is stirred with one molar equivalent of the potassium salt of a dibasic acid or two molar equivalents of the potassium salt of a monobasic acid in an aqueous medium.