Patent Publication Number: US-4481218-A

Title: 3-(2,2,2-Trimethylhydrazinium)propionate and method for the preparation and use thereof

Description:
This is a continuation of application Ser. No. 194,295 filed as PCT SU79/00060, Jul. 27, 1979, published as WO80/01068, May 29, 1980 §102(e) date July 27, 1980. 
    
    
     FIELD OF THE INVENTION 
     The present invention relates to a novel compound out of the class of novel chemical compounds--linear hydrazinium betaines--3-(2,2,2-trimethylhydrazinium)propionate, a method for preparing same and use as an active principle for a growth stimulator for fowl and animals. 
     BACKGROUND OF THE INVENTION 
     Compound having a betaine structure with a hydrazinium group as a positively charged grouping have not been hitherto described in the literature, 
     The compound according to the present invention, viz. 3-(2,2,-trimethylhydrazinium)propionate possessing this structure is novel and hitherto unknown from the literature. 
     Known in the art is the use, as growth stimulator for farm animals and fowl, of antibiotics such as chlorotetracycline, nitrofuran preparations, arsenic derivatives, enzymes, hormones, tissue preparations, phosphatides and the like. However, antibiotics and nitrofuran preparations are efficient only under conditions of infection. 
     On the other hand, substantially all of the known growth stimulators: antibiotics, enzymatic preparations, are economically inefficient. Some of these such as arsenic preparations, are toxic. In cases of the use of antibiotics it is necessary to take into consideration the real risk of origination of resistant, bacteria strains, such as pathogenic strains e.g. E.coli. Residual amounts of antibiotics in meat products and eggs can cause allergic diseases. 
     U.S. Pat. No. 2,925,342, published 02.16.60, Cl. nat. C I.99-2. 
     U.S. Pat. No. 3,017,272 published 01.16.62, Cl.nat. C I.99-2; U.S. Pat. No. 3,639,595 published 02.01.72, Cl.Int. A 61 k 21/00; 
     French Pat. No. 2,137,731 published 02.02.73, Cl.Int. A 23 k 1/00; FRG Pat. No. 1,692,477 published Oct. 19, 1972, Cl. 53 g 4/04). 
     SUMMARY OF THE INVENTION 
     According to the present invention, the novel chemical compound 3-(2,2,2-trimethylhydrazinium)-propionate has the following formula: 
     
         (CH.sub.3).sub.3 N.sup.+ NHCH.sub.2 CH.sub.2 COO.sup.-.2H.sub.2 O (1) 
    
     The method for preparing said compound 3-(2,2,2-trimethylhydrazinium)-propionate comprises passing a solution of 3-(2,2,2-trimethylhydrazinium)alkylpropionate of the formula: 
     
         X.sup.- (CH.sub.3).sub.3 N.sup.+ NHCH.sub.2 CH.sub.2 COOR 
    
     wherein X is Cl, Br, I, CH 3  SO 4 , R is a lower alkyl through a column with a strongly basic anion exchange resin, followed by isolation of the desired product. 
     It is advisable to use a solution of 3-(2,2,2-trimethylhydrazinium)alkylpropionate in water or in a polar organic solvent. 
     As the polar organic solvent use is preferably made of methanol or ethanol. According to the present invention, the growth stimulator involving an active principle and a vehicle contains, as the active principle, 3-(2,2-trimethylhydrazinium-propionate. It is preferred to use the growth stimulator for animals and fowl with a content of the active principle of from 0.001 to 0.5% by weight. 
     DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS 
     The compound according to the present invention, viz. 3-(2,2,2-trimethylhydrazinium)propionate comprises a colourless crystalline powder with a melting point of 254°-256° C. (from ethanol). It is very readily soluble in water, well soluble in ethanol and methanol. The PMR in deuterized water reveals the presence of the following signals τ (p.p.m.): 6.70 singlet, (CH 3 ) 3  N; 6.89, triplet, CH 2 , 
     The toxicological study of 3-(2,2,2-trimethylhydrazinium)-propionate on white mice weighing 20-25 g has shown that with a single peroral administration to the animals in doses of up to 23,000 mg/kg the preparation causes no death of the animals. When administered intraperitoneally, doses of up to 10,000 mg/kg of 3-(2,2,2-trimethylhydrazinium)propionate proved to be non-toxic, No pathological changes in vitally important organs are observed upon administration of the preparation to white rats weighing 200-250 g perorally in doses of 100 mg/kg every day within the period of 3 months. 
     The study of specific activity of 3-(2,2,2-trimethylhydrazinium)-propionate was effected on various animals: mice, rats, pigs, cattle, chickens, goslings, turkey poults and ducklings. The growth-controlling activity of this compound relative to plats has been studied on barley, oats, rye and wheat. 
     It has been found that 3-(2,2,2-trimethylhydrazinium)-propionate possesses a clearly pronounced growth-stimulating activity relative to animals and fowl. Depending on the variety of animals and fowl, the compound according to the present invention ensures a daily weight gain of 8-24%. At the same time, in the test animals and fowl, metabolism is improved and the rate of feedstock consumption is reduced by 7-15% per weight gain unit. 
     In the investigation of the activity of 3-(2,2,2-trimethylhydrazinium)-propionate on plants it has been found that this compound possesses a clearly pronounced growth-controlling effect. This compound ensures a 15% increase in the stem diameter of crops such as oats, barley, rye, wheat, thus improving their stability against lodging. 
     Furthermore, specific gravity of grain crops is increased, thus causing, in general, a yield gain of these crops by 15-30%. 
     3-(2,2,2-Trimethylhydrazinium)-propionate comprises the active principle of a growth-stimulator for animals and fowl. This compound can be used with any vehicle suitable for feeding to animals and fowl, such as water, milk, as well as in a mixture with a feedstuff or any other suitable filler. The growth stimulator for animals and fowl is administered daily in a single dose or several times per day. The efficiency of the growth stimulant does not substantially depend on the mode and number of administrations and is defined mainly by the dose and duration of feeding. The growth stimulant is employed for different kinds of animals and fowl and a preferred content of the active principle is varied within the range of from 0.001 to 0.5% by weight, or 0.1-100 mg per kg of the animal body weight. 
     The compound according to the present invention, viz. 3-(2,2,2-trimethylhydrazinium)-propionate is produced by passing a solution of a 3-(2,2,2-trimethylhydrazinium)-alkylpropionate of the general formula: 
     
         X(CH.sub.3).sub.3 N.sup.+ NHCH.sub.2 CH.sub.2 COOR, 
    
     wherein X is Cl, Br, I, CH 3  SO 4 , R is a lower alkyl, through a column with a strongly-basic anion exchange resin, followed by isolation of the desired product. The nature of the anion does not affect the yield and quality of the desired product. Use is made mainly of salts of 3-(2,2,2-trimethylhydrazinium)alkylpropionates, wherein the alkyl radical is methyl, ethyl, propyl or other lower alkyl radicals. The specific feature of the method according to the present invention for the preparation of 3-(2,2,2-trimethylhydrazinium)-propionate resides in that passing a solution of a salt of 3-(2,2,2-trimethylhydrazinium)-alkylpropionate through a column with a strongly-basic anion exchange resin such as Amberlite IRA-400 or another strongly-basic resin causes hydrolysis of the estereal grouping simultaneously with exchange of the acid anion to hydroxy ion, thus considerably reducing the synthesis duration. In the process according to the present invention use can be made of any suitable solvent such as water, a polar organic solvent (alcohols and the like) so that the solvent nature does not substantially affect the yield and purity of the desired product. 
    
    
     For a better understanding of the present invention some specific examples illustrating the method for preparing the novel compound 3-(2,2,2-trimethylhydrazinium)-propionate and use are given hereinbelow. 
     EXAMPLE 1 
     A solution of 196.5 g (1 mole) of 3-(2,2,2-trimethylhydrazinium)-methylpropionate chloride in 500 ml of water is passed through a column with the ion-exchange resin Amberlite IRA-400. The solvent is evaporated, the residue is crystallized from ethanol. The yield of 3-(2,2,2-trimethylhydrazinium)-propionate is 140 g (85%); the product comprises colourless crystals melting at 254°-256° C. The PMR-spectrum (τ, p.p.m.): 6.70, singlet, (CH 3 ) 3  N +  ; 6.89, triplet, CH 2  ; 7.77, triplet, CH 2 . Found, %: C 39.56; H 10.10; N 15.36; C 6  H 14  O 2  N 2 .2H 2  O. Calculated, % C 39.56; H 9.89; N 15.30. 
     EXAMPLE 2 
     A solution of 272 g (1 mol) of 3-(2,2,2-trimethylhydrazinium)-methylpropionate methylsulphate in 500 ml of methanol is passed through a column filled with an ion-exchange resin. The solvent is evaporated, the residue is crystallized from ethanol. The yield of the desired product is 148 g (90%) in the form of colourless crystals. The product characteristics are similar to those described in the foregoing Example 1. 
     EXAMPLE 3 
     A solution of 210.5 g (1 mol) of 3-(2,2,2-trimethylhydrazinium)-ethylpropionate chloride in 800 ml of ethanol is passed through a column with an ion-exchange resin (Amberlite IRA-400) and the solvent is evaporated. There are obtained 146 g (about 88%) of 3-(2,2,2-trimethylhydrazinium)-propionate as colourless crystals. The product characteristics are similar to those described in Example 1. 
     EXAMPLE 4 
     A solution of 244.9 g (1 mol) of 3-(2,2,2-trimethylhydrazinium)-ethylpropionate bromide in 500 ml of water is passed through a column with the ion-exchange resin Amberlite IRA-400, then the solvent is evaporated to give 147 g (about 87%) of 3-(2,2,2-trimethylhydrazinium)-propionate as colourless crystals. The product characteristics are similar to those of Example 1 hereinabove. 
     EXAMPLE 5 
     A solution of 277 g (1 mol) of 3-(2,2,2-trimethylhydrazinium)-methylpropionate iodide in 500 ml of water is passed through a column with the ion-exchange resin Amberlite IRA-400, whereafter the solvent is evaporated to give 140 g (85%) of 3-(2,2,2-trimethylhydrazinium)-propionate as colourless crystals. The product characteristics are similar to those described in Example 1. 
     EXAMPLE 6 
     The growth stimulator for animals and fowl containing 3-(2,2,2-trimethylhydrazinium)-propionate as the active principle is tested for efficiency. The test is conducted in chicks of 6 days of age, the test group consisting of 50 chicks and the control group--50 chicks. The test chicks were fed with a feedstuff containing 0.02% by weight of the active principle, the control group--only basic diet without the addition of the active principle. The chicks&#39; diet contained, % by weight: barley--20, wheat--6, corn--40, sunflower and soya-bean grist--19, fish flour--7, meat-bone flour--3, yeast--5. One kilogram of the feedstuff contained 1.25 feeding unit, 3,135 kCal of exchange energy, 21% of a crude protein, 4% of crude cellulose and 197 g of digestable protein. The experiment was carried out for 34 days and by its end the chicks&#39; weight was 440.5 g on the average in the test group vs. 412.2 g of the average weight of the control group chicks, i.e. the chicks fed with 0.02% by weight of the feedstuff of the additive 3-(2,2,2-trimethylhydrazinium)-propionate had a weight gain of 7.7% compared to the control chicks. The test results are shown in Table 1. 
     
                       TABLE 1                                                     
______________________________________                                    
                                     Per-                                 
                            Average  cen-                                 
Feedstuff   Average body weight                                           
                            gain for tage                                 
charac-     of chicks, g, aged:                                           
                            the test to the                               
No.  teristic   6 days  30 days                                           
                              40 days                                     
                                    time   control                        
______________________________________                                    
1.   Without    42.7    274.0 412.2 364.5  100.0                          
     special                        ± 10.1                             
     additives                                                            
     (control)                                                            
2.   With the   47.8    289.4 440.5 392.7  107.7                          
     addition                       ± 9.9                              
     of 0.02%                                                             
     by weight                                                            
     of 3-(2,2                                                            
     2-trimethyl-                                                         
     hydrazinium)-                                                        
     propionate                                                           
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     EXAMPLE 7 
     The experiment for testing efficiency of the growth stimulant for animals and fowl according to the present invention was carried out following the procedure similar to that described in the foregoing Example 6; added to the feedstuff was 0.04% by weight of the active principle based on the feedstuff weight. By the end of the 34-days&#39; period the chicks of the test groups fed with the growth stimulant were 12.9% heavier. The test results are shown in the following Table 2. 
     
                       TABLE 2                                                     
______________________________________                                    
                        Average  Per-                                     
           Average body weight                                            
                        mass     cen-                                     
           of chicks, g. aged:                                            
                        gain for tage                                     
     Feedstuff   6       30    40   the test                              
                                           to the                         
No.  characteristic                                                       
                 days    days  days time, g                               
                                           control                        
______________________________________                                    
1.   Without addi-                                                        
                 54.0    261.9 364.3                                      
                                    313 ± 3                            
                                           100.0                          
     tive (control                                                        
2.   With the addi-                                                       
                 51.0    268.9 404.8                                      
                                    353 ± 8                            
                                           112.9                          
     tion of 0.04%                                                        
     by weight of                                                         
     3-(2,2,2-tri-                                                        
     methylhydra-                                                         
     zinium)-propio-                                                      
     nate                                                                 
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     EXAMPLE 8 
     The efficiency of the growth stimulant for animals according to the present invention was tested in feeding of microbless animals. The test was carried out in microbless pig mini-sucklings grown without under sterile conditions in a comparison with pig mini sucklings grown keeping to the requirements of sterile conditions, since birth and to the age of two-months, a feedstuff containing 0.04% by weight of the active principle. Their growth was compared to that of 10 sucklings grown without keeping to sterility conditions and the development of 7 microbless pig sucklings. By the end of the experiment the test pig sucklings were 17.2% heavier than the microbless pig sucklings given no growth stimulant and 18.2% heavier than the sucklings grown under nonsterile conditions and given no growth stimulator. The test results are shown in the following Table 3. 
     
                       TABLE 3                                                     
______________________________________                                    
                    Animal weight                                         
                                Number of ani-                            
                    at the 2-mon-                                         
                                mals in the                               
No.   Animal category                                                     
                    ths&#39; age, g group                                     
______________________________________                                    
1.    Ordinary pig suck-                                                  
                    3,244 ± 335                                        
                                10                                        
      lings given no                                                      
      growth stimulator                                                   
2.    Microbless mini-                                                    
                    3,271 ± 385                                        
                                7                                         
      pigs (sucklings)                                                    
      given no growth                                                     
      stimulant                                                           
3.    Microbless mini-                                                    
                    3,833 ± 348                                        
                                5                                         
      pigs fed with                                                       
      the growth stimu-                                                   
      lator in the amo-                                                   
      unt of 0.04% by                                                     
      weight of the ac-                                                   
      tive principle                                                      
      (based on the                                                       
      feedstuff)                                                          
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     EXAMPLE 9 
     The efficiency of the growth stimulator was tested on white-breed young pigs deprived of mother&#39;s milk with the body weight of about 20 kg, 10 kgs, in each group. 
     The young pigs were in the same pig sty and serviced by one operator; all of the animals were on the same diet consisting of barley flour, green mass of a vetch-oat mixture and whey. The animals were fed two times a day with free watering. To the feedstuff of the test group there were added 200 mg of 3-(2,2,2-trimethylhydrazinium)propionate per 1 kg of dry substance of the feed-stuff (0.02% by weight of the active principle). It was found that the pigs given the growth stimulator for 122 days had a weight gain by 7.4 kg higher as compared to the control group animals, while the consumption of feeding units per every kilogram of the weight gain is by 7.1% less. The test results are shown in Table 4. 
     
                       TABLE 4                                                     
______________________________________                                    
                 Group of animals                                         
Characteristic     Test      Control                                      
______________________________________                                    
Number of animals  10        10                                           
Average body weight by the                                                
                   218 ± 1.5                                           
                             20.0 ± 1.67                               
test beginning, kg                                                        
Average body weight by the                                                
                   88.2 ± 4.0                                          
                             74.0 ± 3.2                                
test end, kg                                                              
Body weight gain during the                                               
                   66.4 ± 3.6                                          
                             59.0 ± 2.1                                
test period, kg                                                           
Average daily weight gain, kg                                             
                   544 ± 32                                            
                             484 ± 17                                  
Average daily weight gain,                                                
                   112.4     100.0                                        
percent of the control                                                    
Feedstuff consumption during                                              
                   270.7     285.5                                        
the test period, feeding                                                  
units                                                                     
Feedstuff consumption per                                                 
                    4.07      4.38                                        
1 kg of the body weight                                                   
gain, feeding units                                                       
Feedstuff consumption per                                                 
                    92.9     100.0                                        
1 kg of the body weight gain                                              
% of control                                                              
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     EXAMPLE 10 
     The efficiency of the growth stimulator was studied on young rats. The experiment was carried out with nondescript young rats weighting about 75 g in groups of 30 animals in each. The growth stimulator was administered perorally to each rat as an aqueous solution at a rate of from 0.1 to 0.001% by weight of the consumed daily diet. The administration of the growth stimulator was effected every day during 10 days. The observation period was 20 days; the rats were weighed after every 4 days. It was found that 3-(2,2,2-trimethylhydrazinium)-propionate in the dose of 0.1% by weight of the consumed daily diet substantially did not affect the weight gain of the rats, whereas the addition of 0.01% and 0.001% by weight of the consumed daily diet increased the body weight gain of the rats by 12 and 12.5% respectively. On completion of the experiment the animals were killed and the parenchymatous organs and peripheral blood were analyzed. No pathology was observed. The test results are shown in Table 5. 
     
                       TABLE 5                                                     
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             Weight on the day of weighing, g                             
             since the beginning of feeding                               
No.   Animal category                                                     
                   4      8    12    16    20                             
______________________________________                                    
1.    Animal given no                                                     
                   74 ±                                                
                          89 ±                                         
                               101 ±                                   
                                     116 ±                             
                                           131 ±                       
      growth stimulator                                                   
                   5      7    6     9     10                             
2.    Animals given the                                                   
                   76 ±                                                
                          95 ±                                         
                                98 ±                                   
                                     107 ±                             
                                           129 ±                       
      growth stimulator                                                   
                   4      6    5     8     6                              
      with 0.1% of the                                                    
      active principle                                                    
3.    Animals given the                                                   
                   79 ±                                                
                          93 ±                                         
                               125 ±                                   
                                     139 ±                             
                                           158 ±                       
      growth stimulator                                                   
                   5      8    10    7     9                              
      with 0.01% of the                                                   
      active principle                                                    
4.    Animals given the                                                   
                   76 ±                                                
                          99 ±                                         
                               131 ±                                   
                                     154 ±                             
                                           164 ±                       
      growth stimulator                                                   
                   5      7    8     8     7                              
      with the content                                                    
      of the active pri-                                                  
      ciple of 0.001%                                                     
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     EXAMPLE 11 
     The growth stimulator efficiency was studied on pubertal rats. The experiment conditions were similar to those described in the foregoing Example 10. It was found that the growth stimulator with the content of the active principle of 0.101% and 0.001% by weight of the dry feedstuff results in the body weight gain of the by 9 and 13.5% respectively. The test results are shown in the following Table 6. 
     
                       TABLE 6                                                     
______________________________________                                    
           Weight, g, on the weighing day                                 
No. Animal category                                                       
             4       8       12    16    20                               
______________________________________                                    
1.Animals given no                                                        
             160 ±                                                     
                     168 ±                                             
                             179 ±                                     
                                   188 ±                               
                                         209 ±                         
growth stimulator                                                         
             11      10      12    9     13                               
2.Animals given the                                                       
             155 ±                                                     
                     161 ±                                             
                             174 ±                                     
                                   187 ±                               
                                         203 ±                         
growth stimulant                                                          
             10      10      8     11    9                                
with 0.1% of the                                                          
principle                                                                 
3.Animals given the                                                       
             169 ±                                                     
                     175 ±                                             
                             189 ±                                     
                                   215 ±                               
                                         229 ±                         
growth stimulator                                                         
             7       8       13    12    11                               
with 0.01% of the                                                         
active principle                                                          
4.Animals given the                                                       
             175 ±                                                     
                     189 ±                                             
                             210 ±                                     
                                   231 ±                               
                                         248 ±                         
growth stimulator                                                         
             11      12      9     14    12                               
with 0.001% of                                                            
the active princip-                                                       
le                                                                        
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     INDUSTRIAL APPLICABILITY 
     The novel compound according to the present invention, i.e. 3-(2,2,2-trimethylhydrazinium)-propionate posesses the ability of controlling plant growth, stimulating growth of animals and fowl, wherefore it is useful in agriculture for increasing yield of crops and stimulation of growth of animals and fowl. The novel compound according to the present invention can be also useful as an intermediate product for the synthesis of polyamide resins.