IL27670A - Antibiotic compositions containing d-cyclo serine and o-carbamyl-d-serine - Google Patents

Description

27670/2 Antibiotic coapoaitione coatainitie I» cyclo serine and O-carbamyl-D-serine This invention relates to new compositions useful in veterinary medicines and as animal feed additives and more particularly to a new composition containing as its essential ingredients an i/v antibiotic such as bacitracin in combination with teycloserine and - O-carbamyl-D-serine.

One important trend in modern production of marketable animals, e.g., poultry, swine, cattle, sheep, etc, , is toward a more rapid development of market-weight animals. Thus the amount of time in which equipment, space, and feed are occupied in grow-0 ing the animal to a marketable size is of importance as is the amount of consumed feed, i.e., feed efficiency. An increased feed efficiency results in less cost in producing market-weight animals β It is known that various antibiotics promote growth and increase feed efficiency of animals. Such antibiotics include bacitracin, neomycin, penicillin, the tetracyclines, the sulfa drugs, tylosin, chloramphenicol, erythromycin, and o!hers. At different dosage levels, these antibiotics also are effective therapeutic agents for various bacterial infections in animals. For example, bacitracin is effective in the prevention of bacter-0 ial swine enteritis , oxytetracycline is effective in the treatment of bacterial diarrhea in sheep and other animals and the other antibiotics are effective against these or other bacterial infections.

It has now been surprisingly discovered that the pharmacological activity in animals of these antibiotics is potentiated by an admixture of^cycloserine and 0-carbamyl,-D-serine. Cycloserine itself is an antibiotic active against many gram-negative and gram- positive microorganisms. It is produced during the cultivation of a particular strain of Streptomyces lavendulae and is described in 0 U. S, patent 2 ,773,878. O-carbamyl-D-se'rine , described in U, S. patent 2,885,133, is also known as a therapeutic agent in veteri- nary medicine.

The potentiation of the pharmacological activity of the / antibiotics is due to the admixture of/ cycloserine and O-carbamyl- D-serineβ Data show that the growth activity of the antibiotic is not improved when either cycloserine or O-carbamyl-D-serine alone are administered with the antibiotic in amounts equivalent to those of the three component mixtures. The activity of the ad- /< mixture of cycloserine and O-carbamyl-D-serine with the antibiotic also surpasses the separate activity of equivalent amounts of the 0 individual constituents of the mixture» D- Antibiotics which are potentiated by an admixture of/cycloserine and O-carbamyl-D-serine include neomycin; bacitracin, including the active salts of bacitracin such as zinc bacitracin, manganese bacitracin and bacitracin methylenedisalicylate , feed grade bacitracin and bacitracin-containing materials such as the dried bacitracin-containing whole beer remaining after separation of bacitracin from fermentation media in which it is produced; penicillin, iaea , from procaine penicillin; the tetracyclines, including tetracycline, chlortetracycline and oxytetracycline ; 0 tylosin; chloramphenicol; erythromycin; and sulfa drugs, including sulfanilamidoquinoxaline , sulfamethoxypyridazine, sulfathiazol , sulfanilamide, sulfamethazine, and sulfadimethoxine ; alone or in combination or in combinations with other materials common to feed additives such as described in the Feed Additive Compendium issued February, 1966, by the Miller Publishing Company,, The amount of antibiotic and admixture of O-carbamyl-D- _ serine and/cycloserine administered is an effective amount, that is, an amount sufficient to produce a pharmacological effect such as to improve the rate of growth and the feed efficiency of the 0 animal, or a therapeutic amount for the particular disorder. The exact amount of the dosage will vary depending upon the particular antibiotic, the intended use, the ratio of O-carbamyl-D-serine /cyclos used, the body weight of the subject, the method used in administering the dose to the subject, and whether single or multiple dosages are contemplated^ The weight ratio of antibiotic to admixture of^cycloserine and O-carbamyl-D-serine will generally fall in the range of about 1:1 to 1:9, preferably about 1:2 to 1:6» The weight ratio of O-carbamyl-D-serine to/ cycloserine in the admixture will usually fall in the range of about 20:1 to 1:4, preferably about 12:1.to 1:2, Ordinarily the dosage of the composition orally administered falls in the range of about 1 to 700 milligrams, preferably 60 to 500 milligrams per kilogram body weight of the animal per day. The antibiotic is ordinarily administered to the animal as a dosage unit comprising from about 1 to 100 grams, preferably 2 to 50 grams of the antibiotic, per ton of feed, i„e., nutrient ration such as a basal feed, as a carrier.

The composition of the invention can also be orally ministered in vehicles well known to the art, for instance, in solution in water, alcohol, oils, syrups, and suspending agents, eeg. , gum arabic or similar polysaccharides „ Alternatively, the composition may be given with solid carriers which can be any palatable foodstuffs such as sugar, sweetened starch compositions and the like. The compositions can be in pill, paste, capsule or tablet form, which can be covered with a sweetened or flavored edible coating, if desiredo In addition to oral administration, the composition of this invention may be administered by any other suitable method including parenteral administration „ For example, the composition may be suspended in a suitable injection suspension such as peanut oil and injected parenterally „ The following examples illustrate the invention, but are not to be considered limiting. Example I illustrates the potentiation of zi .nc baci.traci.n by a mi.xture of/pc-ycloserine and O-carbamyl-D-serine when treating healthy swine, EXAMPLE I Two litters (16) of apparently healthy, well doing pigs were obtained, weaned at 8 weeks, ear-notched, vaccinated for cholera and erysipelas and allocated to two test groups on an equalized weight and sex basis0 Individual weights and group fe consumption were recorded every two weeks. The two treatment groups were a control group and a test group of eight pigs each. The control group was fed the basal ration of Table I in self-feeders. The test group was fed the basal ration mixed with an admixture of 50 grams of zinc bacitracin and 200 grams of/cyclo- i serine ( CS ) and O-carbamyl-D-serine (OCS) in a weight ratio of CS:0CS of 1:3, The zinc bacitracin employed was the product obtained by drying the beer remaining following precipitation of zinc bacitracin from fermentation media containing the same and contained 10 grams of zinc bacitracin per pound, TABLE I Ground Yellow Corn 795 lb.

Soybean Oil Meal-50% 180 lb.

Ground Limestone 10 lb.

Di Calcium Phosphate 10 lb.

Iodized Salt 5 lb„ Trace Mineral Mix 227 gm.

Vitamin A (30,000 IU/gm. ) 50 gm.

Vitamin D2 (75,000,000 IU/gm,) 0,75 gm.

BY-24 16, d gm.

Calcium Pantothenate 5,6 gm.

Niacin 98% 8,0 gm.

Choline Chloride 25% 200,0 gm.

Proferm 12 302 , 0 gm. 1,001, 8 lb.

The addition of zinc bacitracin and CS:OCS to the basal ration of healthy pigs produced a 32% increase in average daily gain over the controls which gained at a rate of 1.51 pounds per day. The results are summarized in Table II.

TABLE II Zinc Bacitracin Basal CS ;0CS No, Pigs 8 8 Av, Initial wt, (Lbs.) 38,4 38.9 Av, Final wt. (Lbs.) 166.9 207.7 Days on test 85 85 Av, Daily Gain (Lbs.) 0-1 week 1.14 1.07 1- 3 weeks 1.35 1.68 3-5 weeks 1,20 1.90 -7 weeks 1.81 2.24 7- 8 weeks 2.00 2, 62 8-10 weeks 1,43 2,26 10-12 weeks 1.73 2.05 0-12 weeks 1.51 1.99 Lbs, Feed/Lb. Gain 0-12 weeks 2.98 3.13 The following example illustrates the effect of zinc bacitracin and OCS alone and in combination on growth and feed efficiency in swine.

EXAMPLE II Forty weanling pigs, 6 to 7 weeks of age and averaging 24 pounds, were allocated to four test groups on an equalized weight basis. All pigs were vaccinated for cholera and erysipelas at the start of the test. They were wormed with piperazine in the water after 6 weeks on test, Individual weights and group feed consumption were recorded every two weeks , Treatment groups included zinc bacitracin at 50 grams per ton, a mixture per ton of zinc bacitracin, 50 grams, and OCS, 200 grams, and OCS at 250 grams/ ton, The zinc bacitracin was that employed in Example I. The basal ration of Table I was used to prepare the feeds,, Test period was 8 weeks.

No growth response was obtained from any of the treatments, A slight (6%) improvement in feed conversion was evident in the group receiving zinc bacitracin alone, The combination of zinc bacitracin and OCS or OCS alone was not effective in promoting growth and zinc bacitracin alone produced improvement only in feed efficiency and not in gain. The test results are summarized in Table III, Basal Zinc Bacitracin + OCS Zin No, of Pigs 10 10 Av, Initial Wt, (Lbs,) 23,9 23,7 Av, Final Wto (Lbs,) 98,6 97,3 Days on Test 56 56 Av. Daily Gain (Lbs,) 0-2 weeks 0„92 0,96 2-4 weeks 1,23 1,18 4-6 weeks 1,50 1,43 6-8 weeks 1,68 1,68 0-8 weeks 1,33 1, 31 ± ,18 Lbs, Feed/Lb, Gain 0-2 weeks 1,84 1,72 2-4 weeks 2.38 2,50 4-6 weeks 2,49 2,52 6-8 weeks 2,81 2,70 0-8 weeks 2,45 2,43 The following example illustrates the efficacy of an admixture of zinc bacitracin, CS and OCS in treating swine affected with rhinitis and virus pneumonia, EXAMPLE III Ten weanling pigs were procured from a herd having a high incidence of rhinitis and virus pneumonia,, Each pig was ear-notched, weighed and started on a basal diet for three days0 Fecal samples were checked, ascarid eggs identified, and the pigs were wormed with piperazine in water. After three days the pigs were reweighed and allocated on an equalized weight basis into a basal and a treatment group of five pigs each. Feed and water were supplied ad-libitum» The pigs were weighed individually every two weeks, and feed consumption was recorded. The treatment group received the basal ration mixed with per ton of ration an admixture of 50 grams of zinc bacitracin and 200 grams CS:0CS (1:3)» The zinc bacitracin was that employed in Example I. Test period was nine weeks.

The addition of the admixture of zinc bacitracin, CS and OCS to the ration of Table I resulted in a 33% improvement in daily gain and a 7% improvement in feed efficiency during the nine week test period as summarized in Table IV„ TABLE IV Basal Zinc Bacitracin + CS:0CS No, Pigs 5 5 Av, Initial wt, (Lbs,) 13,5 13,7 Avo Final wt0 (Lbs0) 55,4 69,2 Days on Test 63 63 Av, Daily Gain (Lbs,) 0-2 weeks ,20 o 32 2-4 weeks ,67 o 79 4-6 weeks ,62 , 77 6-8 weeks 068 1.21 8-9 weeks 1, 08 1,74 0-9 weeks «,66 , 88 Lbs, Feed/Lb, Gain 0-2 weeks 3.09 3 18 2-4 weeks 2 „ 05 1 96 4-6 weeks 3,12 2 99 6-8 weeks 2,72 2 49 8-9 weeks 2,49 2 19 0-9 weeks 2,64 2 47 At completion of the test the treated group was fed the basal diet for 48 hours and then all pigs were necropsied.

Necropsy revealed extensive lung damage in all basal fed animals with evidence of active pneumonic lesions , Treatment pigs showed evidence of early infection and healed lesions with no active areas present. Nasal turbinates were extensively damaged or nonexistent in most all pigs, The following example illustrates the effect of zinc bacitracin and CS:0CS alone and in combination in pigs affected with rhinitis and virus pneumonia,, EXAMPLE IV Forty weanling pigs were obtained from a herd of swine having a high incidence of rhinitis and virus pneumonia. They were ear-notched, weighed and allocated to four treatment groups of 10 pigs each on an equalized weight basiso Individual weights and group feed consumption were recorded every two weeks. After two weeks all pigs were wormed with piperazine in the water and vaccinated for cholera and erysipelas. All pigs were fed basal diet for 48 hours before the completion of the test and the final weigh period. The four treatment groups were (1) fed the basal ration of Table I as a control; (2) the basal ration, zinc bacitracin, CS , OCS feed mixture of Example I; (3) zinc bacitracin in the amount of 250 grams per ton of the basal ration of Table I; and (4) the basal ration of Table I admixed with 250 grams per ton of ration of a mixture of CS and OCS in a weight ratio of CS:0CS of 1:3, The test period was 12 weeks.

The combination of zinc bacitracin and CS:0CS produced a 21% increase in average daily gain over the 12 week period. Zinc bacitracin alone or CS:0CS alone gave no response. The test re-suits are set forth in Table V, Control Zinc bacitracin + CS:0CS Zinc No. Pigs 8 9 Av. Initial Wt, (Lbs0) 15.3 14.7 Av, Final Wt. (Lbs, ) 84, 6 100, 7 Days on Test 84 84 , Daily Gain (Lbs,) 0-2 weeks .31 54 2-4 weeks ,30 47 4-6 weeks ,72 75 6-8 weeks 1,02 21 8-10 weeks 1,25 54 -12 weeks 1,24 34 0-12 weeks ,82 99 Lbs. Feed/Lb, Gain 0-2 weeks 2 77 06 2-4 weeks 3 49 05 4-6 weeks 2 65 10 6-8 weeks 2 51 29 8-10 weeks 2 64 49 -12 weeks 2 94 24 0-12 weeks 2 76 70 Five pigs from each treatment group were necropsied? At necropsy all pigs showed evidence of rhinitis and virus pneumonia to varying degrees, The following example illustrates the effect of CSsOCS on growth promotion and feed efficiency in swine with rhinitis-virus pneumoniae EXAMPLE V Thirteen weanling pigs obtained from a herd severely affected with rhinitis and virus pneumonia were allotted to weight equalized basal and treatment groups of six and seven pigs respectively and fed and watered ad-libitum. Pigs were weighed individually every two weeks and feed consumption was recorded, The treatment group received the basal ration of Table I with CSsOCS (Is 3) admixed therewith in the amount of 250 grams per ton. The test period was eight weeks, CS:0CS (1:3) added to the ration had no effect on rate of gain or feed efficiency of pigs affected with rhinitis and virus pneumonia. Two very poor pigs receiving CSsOCS lived through the test period while a similar pig in the basal group died. Analysis of variance of the data show no significance nor is there any percentage difference, CS:0CS (1:3) alone is not effective in promoting growth in pigs with rhinitis and virus pneumonia.

TABLE VI BASAL No, Pigs 5 Av, Initial wt, (Lbs,) 16 o 6 Av. Final wt, (Lbs,) 82 , 6 Days on Test 56 Av. Daily Gain (Lbs,) 0-2 weeks o 72 2-4 weeks , 91 4-6 weeks 1, 35 6-8 weeks lo 63 0-8 weeks 1, 13 , Feed/Lb, Gain 0-2 weeks 1, 95 2—- weeks 2 , 23 4-6 weeks 2 o 27 6-8 weeks 2 , 22 0-8 weeks 2 , 18 '"'Figures in parenthesis are those obtained when two small pigs are compensate for similar pigs in the control group which died.

At the completion of the test the CS:OCS group received basal diet for 48 hours prior to necropsy,, Necropsy revealed only moderate damage to the turbinates and lungs of the pigs in both groups o The following example illustrates the efficacy of an admixture of CS , OCS, sulfadimethoxine and zinc bacitracin in swine from a herd infected with rhinitis-virus pneumonia, EXAMPLE VI The test compared two groups of five pigs each allocated on an equalized weight basis, one of the groups receiving the basal ration of Table I and the other group receiving the basal ration having admixed therewith per ton of ration an admixture of 50 grams zinc bacitracin, 50 grams CS and OCS at a CS:0CS ratio of 1:3, and 100 grams sulfadimethoxine „ All pigs were affected with rhinitis and virus pneumonia. The pigs were fed ad-libitum, were weighed individually every two weeks , and feed consumption for each group was recorded at the weigh period..

The addition of the zinc bacitracin, CS , OCS and sulfa-dimethoxine admixture to the normal basal ration improved the growth rate for six weeks by 14% and the feed efficiency by 7% in pigs affected with rhinitis and virus pneumoniae Bac,-CS:OC Basal Sulfa Daily Gain ( Lbs , ) 0-2 weeks .73 1.08 2-4 weeks 1.08 1.26 4-6 weeks 1.42 1, 35 0-6 weeks 1,08 1.23 o Feed/Lb, Gain 0-2 weeks 3.11 67 2-4 weeks 2.93 33 4-6 weeks 2,28 52 0-6 weeks 2,69 50 Avo Initial t. (Lbs,) 43.4 42,7* ( Av, Final wt, (Lbs,) 88,6 98,7 Days on Test 42 42 No0 Pigs 5 4 *0ne pig died of pneumonia after 4 weeks, —Final weight on four pi EXAMPLE VII Four separate 2 0 0 gram batches of a mixture of CS2OCS in weight ratios of 1 : 9 , 1 : 1 and 3 : 1 , respectively, each batch admixed with 5 0 grams of the zinc bacitracin of Example I are prepared and each of the four admixtures is separately admixed with a ton of the basal ration of Table I„ Each feed admixture is administered to swine „ The rate of growth and feed conversion of the swine are improved. The improvement is noted with healthy swine as well as diseased swine, EXAMPLE VIII A two hundred gram batch of a mixture of CSsOCS in a 1 : 3 weight ratio is separately admixed with fifty grams of each of the following antibiotics: bacitracin; manganese bacitracin; bacitracin-methylene disalicylate ; neomycin; tylosin; penicillin (from procaine penicillin); tetracycline; oxytetracycline ; chlortetracycline ; chloramphenicol; erythromycin; sulfamethazine; sulfanilamide; sulfamethoxypyridazine ; sulfathiazole ; sulfanil-amidoquinoxaline ; sulfadimethoxine ; zinc bacitracin admixed with penicillin (from procaine penicillin) (containing between 5 0 and 7 5 % zinc bacitracin); tylosin admixed with penicillin (from procaine penicillin) ( 1 „ 2 parts penicillin to 2 parts tylosin); and chlortetracycline-sulfamethazine-penicillin in a weight ratio of l:l: 0 o 5 o Each admixture is separately admixed with a ton of the basal ration of Table I» EXAMPLE IX The antibiotic admixtures with CS and OCS of Examples VII and VIII are admixed with nutritionally adequate rations for chickens, turkeys, sheep and cattle and administered as feedo The rate of growth and feed conversion of the animals is improved, This improvement is noted with healthy animals as well as diseased animals The /cycloserine and O-carbamy1-D-serine employed in the above examples were high potency pharmaceutical grade products „ D- Other less pure forms of/cycloserine and O-carbamyl-D-serine can be employed, however, such as the dried crude/ cycloserine whole beer, or the dried crude /cycloserme-containmg broth obtained after filtration of the mycelium from the fermenter effluent described in U o S c patent No, 2 , 7 7 3 , 8 7 8 , or the corresponding dried O-carbamyl-D-serine containing filtrate described in U „ S o patent oo 2 , 8 8 5 , 4 3 3 ο In any event, the antibiotic-¾ycloserine-O-carbamyl-D-serine admixture should be maintained as dry as possible to preserve its potency and when admixed with nutritionally adequate rations, the rations should have low water content, i.e., below about 10% by weight of water, advantageously below about 5 % by weight of water- By the term "antibiotic" is meant any drug intended for use by man containing any quantity of any chemical substance which is produced by a micro-organism and which has the capacity to inhibit or destroy microorganisms in dilute solution (including the chemically synthesized equivalent of any such substance) and in addition the term is construed to include those materials having chemotherapeutic activity, such as the sulfa drugs, normally employed in animal nutrition or animal disease therapy „

Claims (2)

1. HAVING NOW particularly described ?Jid ascertained the nati of our said invention and in what manner the same is to be performed^ we declare that what we claim ist The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows : 1» A composition comprising an admixture of an antibiotic ^ 1 other than a/c-ycloseri .ne or O-carbamyl-D-serm . e and an anti .bi.oti.c ,D- ^ . potentiating amount of a mixture of/ cycloserine and O-carbamyl-D- serine .
2. 2. A composition comprising an admixture of at least one , , animal feed additive antibiotic other than /cycloserine or O-carbamyl- D-serine and an antibiotic potentiating amount of a mixture of D- cycloserine and O-carbamyl-D-serine; the weight ratio O-carbamyl- D-serine toD//c-ycloserine being within the range of about 20 :1 to 1:4. 3o The composition of claim 2 wherein the weight ratio of Ί O-carbamyl-D-serine to/cycloserine is within the range of about 12 :1 to 1:2. 4» A composition comprising zinc bacitracin and a potentiat- /D- j rj mg amount of a mixture of/cycloserine and O-carbamyl-D-serme . 5<, A composition comprising zinc bacitracin and a potentiat- - ing amount of a mixture of/cycloserine and O-carbamyl-D-serine , the weight ratio of O-carbamyl-D-serine to cycloserine being within the range of about 20 :1 to 1:4» 6, The composition of claim 5 wherein the weight ratio of Dr . . . O-carbamyl-D-serine to/cycloserine is within the range of about 12 :1 to 1.2. 7o A method for improving the rate of growth and feed conversion of animals comprising administering to said animals an amount of the compositions of claim 1 effective to improve the rate of growth and feed conversion. 8a A method for improving the rate of growth and feed conversion of animals comprising administering to said animals an amount of the two- composition^ of claim 4 effective to improve the rate of rowth and feed conversion,,