Animal feed compositions useful as growth promotors and for reduction of fat in animals,comprising phenylethanolamine derivatives and certain such novel compounds

Substitution products of certain 1-(amir.odihalc. phenyl)-2-aninoethanes and the acid addition salts thereof are disclosed in United States Patent 3,536,712, (corresponding to local patent No.28662) issued on 27th October, 1970. Said Israeli Patent No. 28662 discloses similar compounds which are useful for enhancing the blood circulation, and as bronchodilators, analgesics, sedatives, 'antipyretics, antiphlogistics and antitussive which is entirely different from the utility of the compounds of this invention.

The U. S. Patent 3,818,101 discloses the use of certain -adrenergic agents for improving the intake of feed in meat-producing animals, whereas the compounds of the instant application generally depresses feed intake.

Specifically, patentees disclose methods for the synthesis’of said compounds and state that said compounds are useful for enhancing the blood circulation, and as bronchodilators, analgesies, sedatives, antipyretics, antiphiogistics and antitussives in warm-blooded animals. Patentees, however, exemplify only the analgesic utility. They do not indicate or suggest that said compounds are useful for lowering the deposition of fat or increasing the growth rate in worn-blooded animals, particularly farm and domestic animals, such as swine, poultry, dogs, sheep, goats, cats or cattle.

It has now been found that the growth rate and the depression of fat deposition of meat-producing animals such as swine, chickens, turkeys, domestic pets, rabbits, sheep, goats and cattle, including calves, can be increased, and the efficiency of feed utilization thereby measurably improved by the oral or parenteral administration to said animals of an effective amount of a compound having the structure:

<img file="IL60530A_D0001.tif" />

9.7.80 wherein X is hydrogen or halogen ( chlorine or bromine); Y Is hydrogen,

NH₂ or nhcor₅; z is h, halogen ( chlorine or bromine) or OH; R^ is hydrogen; R₂ is hydrogen, alkyl (straight or branched-chain) or₍%^*C₄ alkenyl; is hydrogen, C^-Cg alkyl (straight or branched-chain), C₃-Cg cycloW 5 alkyl, methoxypropyl, ^c3־^c4 alkenyl, phenyl, 2-hydroxyethyl, a-a-dimethylphenethyl or benzyl; and when R₂ and R₃ are taken together with the nitrogen to which they are attached, they may represent morpholino or N'-C^-C^ alkyl9.7.00 piperazine; R₄ is hydrogen, hydroxyl or OR₆; R₅ is

9.7.80 10 C^-C₄ alkyl; Rg is C-^-Cg alkyl; with the provisos that when R₃ is phenyl, 2.hydroxyethyl, a,a-dimethy!phenethyl, cycloalkyl C^-Cg, benzyl or methoxypropyl, R₂ is hydrogen;

£ and when Z is OH, X and Y are hydrogen; and when Y is NHCORg,

- at least one of X and Z is hydrogen; and provided also that 15 at least one of X,Y and Z represents a substituent other than hydrogen; racemic mixtures of the above-identified compounds and the optically active isomers and non-toxic, pharmacologically acceptable acid addition salts thereof.

Preferred compounds for use in the method of this 20 invention have the above structure wherein X and Z are each chlorine or bromine; Y is hydrogen or NH₂; Rj is hydrogen or ^C1’^C4 ³³kyl׳ ^r4 ^is hydroxyl; or a non-toxic, pharmacologically acceptable acid addition salt thereof. Some of the said compounds are novel and are claimed as such.

The most preferred compounds for use in enhancing the growth rate of meat-producing animals and for improving the efficiency of feed utilization thereby are: 4-amino-a[(tert-butylamino)methyl]-3,5-dichlorobenzyl alcohol hydrochloride; 4-amino-3,5-dibromo-a-[(diisopropylamino)methyl]״ benzyl alcohol hydrochloride; 4-amino-3,5-dichloro-a-[(di30 isopropylamino)methyl]benzyl alcohol hydrochloride; 4amino-3,5-dibromo-a-[(tert-butylamino) methyl]-benzyl alcohol hydrochloride; 4-amino-3,5-dichloro-a-[(methyLamino)methyl]benzyl alcohol hydrochloride; 4-amino-3,5-dichloro-a-[(allylamino)methyl]benzyl alcohol; 4-amino-3-bromo-a-[(tert35 butylamino)methyl]-5-chlorobenzyl alcohol hydrochloride;

a- [4-amino-3, 5-dichlorophenyl] -4-morpholineethanol hydrochloride; 4-amino-3-bromo-a-[(tert-butylamino)methyl]-5-360530/2 chlorobenzyl alcohol hydrochloride and a-[(tert-butylamino) methyl]-3,5-dichlorobenzyl alcohol hydrochloride.

It is found, that formula (I) compounds below (wherein Y is hydrogen) can be prepared by the condensation of an appropriately substituted sytrene oxide with the appropriately substituted amine in the presence of an inert solvent, such as a lower alcohol at or near the boiling point of same, as shown below:

<img file="IL60530A_D0002.tif" />

wherein X and Z are halogen, R₂ and Rg are as hereinabove defined and Y is hydrogen. Thus, 3,5-dichlorostyrene oxide can be reacted with an equimolar or molar excess of t-butylamine in ethanol at reflux from about 1 to about 8 hours, or until the reaction is essentially complete and the desired a-[(t-butylamino)methyl]-3,5-dichlorobenzyl alcohol is obtained as illustrated below:

<img file="IL60530A_D0003.tif" />

The thus obtained product can be purified by known procedures, such as chromatography or recrystallization of salt ־.hereof.

/ί

The thus obtained halo compound is isolated by conventional laboratory methods and is then reacted with the appropriate alcohol, under an inert blanket of gas, such as nitrogen at a temperature range of from about 0 to 5°C. Thus thus obtained formula (I) product is then isolated by standard laboratory methods and purified, if so desired. The above reaction sequence may be graphically illustrated as follows:

X ¹⁰ /7SOCK

Y-C p-&lt;pi־CH₂NR₂R₃

OH

Z

X

Y_// 7—CH-CH-NR-R- 6?>

\=/ A! z

20X γ __/7 ׳-CH-CH-NR-R.,

Vi., z wherein X,Y.Z,R₂׳R2 and Rg are as hereinabove defined.

Alternatively, a formula (I) compound wherein

R₄ is ORg may be prepared by dissolving the corresponding formula (I) compound wherein R^ is OH in the corresponding RgOH alcohol and saturating the thus obtained solution with dry HC1 gas. The reaction mixture is then stirred at room temperature for a period of time sufficient to essentially complete the reaction and the product is then isolated by standard laboratory procedures and purified, if so desired. This reaction sequence may be illustrated as follows:

mjw/1

<img file="IL60530A_D0004.tif" />

R,OH

CH~CH-NR_R --2----| 223 HC1 gas

OH

<img file="IL60530A_D0005.tif" />

״herein X,Y,־,R₂,R₃ ״־* ^R_S^{ara as bareinabove dafined}.

In the present specification and claims the term a,a-dimethy!phenethyl means a structure having the following configuration:

<img file="IL60530A_D0006.tif" />

When orally administered in the feed, generally about 0.01 to 300 grams per ton of feed of the aboveidentified phenylethanolamine derivative or acid addition salt thereof, is effective for enhancing the growth rate and improving the efficiency of feed utilization by the above-mentioned meat-producing animals.

־ 6.Since the effective and preferred dietary levels of the active ingredient vary somewhat from species to species in the above-mentioned animals said level's for each animal species are listed in Table I below on a gram per ton of feed basis:

Table I

<td> Compound</td><td> Effective Feed Level g/ton</td><td> Preferred Level g/ton</td><td> Animal</td>

<td rowspan="5"> Formula (I)</td><td> 0.1 -300</td><td> 1-200</td><td> Swine</td>

<td> 200־0.1</td><td> 1-100</td><td> Sheep, Goats</td>

<td> 0.01-50</td><td> 10־0.1</td><td> Chickens, Rabbits</td>

<td> 0.01-50</td><td> 10־0.1</td><td> Turkeys</td>

<td> ‘0.1 -200</td><td> 100־1</td><td> Cattle</td>

Animal feed compositions which will provide the desired growth promotion and feed efficiency in the above-mentioned animals can be prepared by admixing the phenylethanolamine derivative or acid addition salt thereof, or an animal feed supplement containing said compound, with a sufficient _ quantity of an appropriate animal feed to provide the desired level of active w J compound in said feed.

Animal feed supplements can be prepared by admixing about 75% to 95% by weight of the phenylethanolamine derivative or acid addition salt thereof, with about 5% to ₂₅ 25% by weight of a suitable carrier or diluent. Carriers suitable for use to make up the feed supplement compositions include the following: alfalfa meal, soybean meal, cottonseed oil meal, linseed oil meal, sodium chloride, cornmeal, cane molasses, urea, bone meal, corncob meal and the like. The carrier promotes a uniform distribution of the active ingredient in the finished feed into which the supplement is blended. It thus performs an important function by ensuring proper distrubition of the active ingredient throughout the feed.

If the supplement is used as a top dressing for feed, it likewise helps to ensure uniformity of distribution of the active material across the top of the dressed feed.

For parenteral administration the phenylethanol$0530/2 amine derivative may be prepared in the form of a paste or pellet and administered as an implant, usually under the skin of the heat or ear of the animal in which enhanced growth rate and/or improved efficiency of feed utilization is 5־ sought.

In practive, parenteral administration generally involved injection of a sufficient amount of the above-said ethane derivative to provide the animal with from 0.001 to 100 mg/kg of body weight of the active ingredient. The pre10 ferred dosage level for swine is 0.01 to 50 mg/kg of body weight and for cattle the range of from 0.001 to 50 mg/kg of body weight of the active phenylethanolamine derivative is preferred. The preferred dose level of said ethane derivative for poultry is about 0.001 to 35 mg/kg of animal 15 body weight and the preferred dose level of said ethanol derivative for sheep and goats is 0.001 to 40 mg/kg of animal body weight. The preferred dose level for rabbits and domestic pets is 0.001 to 35 mg/kg of animal body weight.

Paste formulations can be prepared by dispersing 20 the active ethanol derivative in a pharmaceutically acceptable oil such as peanut oil, sesame oil, corn oil or the like.

Pellets containing an effective level of the phenylethanolamine derivative can be prepared by admixing the above-said active ingredient with a diluent such as carbowax, 25 biodegradable polymers, carnuba wax, or the like. A lubricant, such as magnesium stearate or calcium stearate may be added to improve the pelleting process if desired.

It is, of course, recognized that more than one pellet may be administered to an animal to achieve the de30 sired dose level which will provide the increased growth rate and/or improved efficiency of feed utilization by said animal. Moreover, it has been found that additional implants may also be introduced periodically during the treatment period in order to maintain the proper drug release rate in 35 the animal's body.

In addition to enhanced growth promotion and improved efficiency of feed utilization by meat-producing animals, the compounds of the present invention have the add _ ₈ _ 6Θ530/2 ed advantage that, at selected levels of administration they depress the deposition of fat in said animals. This biological response has substantial advantage to poultrymen and swine producers since administration of said compounds at selected levels yields leaner animals which command premium prices from the meat industry.

The invention has several advantages for the pet owner or veterinarian who wishes to trim unwanted fat from pet animals. For poultry men and swine raisers, using the method of the present invention were attained increased yields of leaner animals which command higher prices from the meat industry. Surprisingly, it is also noted that feed efficiency and animal growth rate are significantly enhanced when the compounds of the present invention are administered to swine and poultry at selected dose levels.

The following examples illustrate the invention.

Example

Evaluation of test compounds as animal growth promoters

CF1 female mice from Carworth Farms are received when they are six weeks old. They are housed ten to a cage in air-conditioned rooms (72®F to 76°F) with automatically controlled lights, 14 hours on and 10 hours off. The basal diet used in these studies is Purina Laboratory Chow (see description below), which is supplied ad libitum. Water is also allowed ad libitum.

Thirteen days after arrival, the mice are weighed in groups of ten and assigned at random to the different treatments. The concentration of the different compounds in the diet is indicated in the following tables. Twelve days later the mice are weighed again and the experiment terminated. Test data are provided in Table II below wherein data are reported as percentage gain over controls. The following is a description of the diet to which the growth-promoting compounds were added.

DIET

Guaranteed Analysis

Crude protein not less than 23.0%

Crude fat not less than 4.5% *0SS0/2

Crude fiber not more than 6.0%

Ash not more than 9.0%

Ingredients Meat and bone meal, dried skimmed milk, wheat germ meal, fish meal, animal liver meal, dried beet pulp, ground extruded corn, ground oat groates, soybean meal, dehydrated alfalfa meal, cane molasses, animal fat preserved with BHA, vitamin B^₂ supplement, calcium pantothenate, choline chloride, folic acid, riboflavin supplement, brewer's dried yeast, thiamin, niacin, vitamin A supplement, D-activated plant sterol, vitamin E supplement, calcium carbonate, dicalcium phosphate, iodized salt, ferrec ammonium citrate, iron oxide, manganous oxide, cobalt carbonate, copper oxide zinc oxide.

TABLE II

Evaluation of Test Compounds as Animal Growth Promoters

<td></td><td> Initial</td><td> Final</td><td></td><td> ί Gain</td><td></td>

<td></td><td> House</td><td> House</td><td> Gain</td><td> Over</td><td></td>

<td> Dosage</td><td> Kt, (ri</td><td></td><td> (grans)</td><td> Control</td><td></td>

<td>¹!Amlno-Q-( tert-butyl-</td><td> •</td><td></td><td></td><td></td><td></td>

<td> aminomethyl)-3,5-dichlorobenzyl</td><td></td><td></td><td></td><td></td><td></td>

<td> alcohol hydrochloride</td><td></td><td> t</td><td></td><td></td><td></td>

<td> (ppm in diet)</td><td></td><td></td><td></td><td></td><td> 1 u</td>

<td> 0</td><td> 23.116</td><td> 21.67</td><td> 1.21</td><td></td><td> 0 I</td>

<td></td><td> 23.46</td><td> 21.19</td><td> 1.03</td><td></td><td> 1</td>

<td> 1</td><td> 22.79</td><td> 21.62</td><td> 1.83</td><td></td><td></td>

<td></td><td> 24.10</td><td> 25.98</td><td> ’ 1.88</td><td></td><td></td>

<td></td><td> 24.23</td><td> 25.52</td><td> 1.29</td><td></td><td></td>

<td></td><td> 23.63</td><td> 21.93</td><td> 1.30</td><td></td><td></td>

<td></td><td> 23.33</td><td> 21.76</td><td> 1.13</td><td></td><td></td>

<td></td><td> 22.75</td><td> 23.86</td><td> 1.11</td><td></td><td></td>

<td> Control Average</td><td> 23.17</td><td> 21.85</td><td> 1.39</td><td></td><td></td>

TABLE II (Continued)

Evaluation of Test Compounds as Animal Growth Promoters

<td></td><td></td><td> Initial</td><td> Final</td><td></td><td> J Gain</td><td></td>

<td></td><td></td><td> House</td><td> House</td><td> Gain</td><td> Over</td><td></td>

<td></td><td> Dosage</td><td> JfLlil</td><td> Ol</td><td> (grama)</td><td> Control</td><td></td>

<td> 50</td><td></td><td> 22.95</td><td> 25.63</td><td> 2.66</td><td></td><td></td>

<td></td><td></td><td> 23.91</td><td> 26.14</td><td> 2.23</td><td></td><td></td>

<td></td><td></td><td> 24.26</td><td> 26.30</td><td> 2.04</td><td></td><td> 1</td>

<td> Average</td><td></td><td> 23.71</td><td> 26.02</td><td> 2.32</td><td> +66.9</td><td> H H</td>

<td> 100</td><td> 23.50</td><td> 25.39</td><td> 1.89</td><td></td>

<td></td><td> 23.80</td><td> 26.04</td><td> 2.24</td><td></td>

<td></td><td> 23.00</td><td> 25.65</td><td> 2.65</td><td></td>

<td> Average</td><td> 23.43</td><td> 25.69</td><td> 2.26</td><td> +62.6</td>

<td> 200</td><td> 23.03</td><td> 24.80</td><td> 1.77</td><td></td>

<td></td><td> 24.50</td><td> 26.12</td><td> 1.62</td><td></td>

<td></td><td> 23.08</td><td> 25.04</td><td> 1.96</td><td></td>

<td> Average</td><td> 23.54</td><td> 25.32</td><td> 1.78</td><td> +28,1</td>

The procedure described above is repeated using control animals for each test. Twelve days after the tests are started the animals are weighed and the test terminated. The results of each test are reported in Table III below as weight gains for each test group and percent gain for each group over controls.

TABLE

Evaluation of Teat Compounds as Animal Growth Promoters

Dosage Gain $ Gain Over

Compound (ppm) (grams) Controls

<td> 4-Amino-3,5־dibroino׳-a*[(tert-butylan!ino)n1ethyll״</td><td> 400</td><td> 16.5</td><td> +22.2</td>

<td> benzyl alcohol hydrochloride</td><td> 200</td><td> 20.4</td><td> +51.1</td>

<td></td><td> 100</td><td> 22.9</td><td> +69.0</td>

<td></td><td> 50</td><td> 23.3</td><td> +?2.6</td>

<td> 44alno-3,5-dlbr0i0T14(dllsopropyla111ino)sethyl|</td><td> 200</td><td> 20.2</td><td> +46.4</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 16.9</td><td> +22.5</td>

<td> P-amino-a-[(dimethylamino)methylI benzyl alcohol</td><td> 200</td><td> 17.6</td><td> +28.3</td>

<td></td><td> 100</td><td> 15.6</td><td> +13.0</td>

<td> P-amlno-a-((dilsopropylaminojmethyl]benzyl alcohol</td><td> 200</td><td> 16.5</td><td> +18.7</td>

<td> hydrochloride</td><td> 100</td><td> 14.2</td><td> + 2.2</td>

<td> *!.A1nino-3|5-dichloro-a-[(dimethylaD1ino)1Dethyllbenzyl alcohol hydrochloride</td><td> 100</td><td> 18.4</td><td> + ?.6</td>

<td> *l“Amin0“3,5-dlchloro-a-[(dii30propylamino)methyll.</td><td> 200</td><td> 23.9</td><td> +66.0</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 21.3</td><td> +47.9</td>

)

TABLE m (Continued)

Evaluation of Test Compounds as Animal Growth Promoters

<td></td><td> Dosage</td><td> Gain</td><td> J Gain Over</td>

<td> Compound</td><td> (Pt&gt;)</td><td> (grams)</td><td> Controls</td>

<td>¹l-Aniino-3,5-dichloro-a7[ (cyclohexylaminolnethyl] -</td><td> ' 200</td><td> 19.3</td><td> +68.5</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 16.3</td><td> +25.6</td>

<td rowspan="2"> P-amlno-a-((tert־butylamino)1nethylIbenzyl alcohol</td><td> 200</td><td> 19.6</td><td> +88.5</td>

<td> 100</td><td> 18,2</td><td> +75.0</td>

<td></td><td> 50</td><td> 17.9</td><td> +72.1</td>

<td></td><td> 25</td><td> 13.6</td><td> +30.8</td>

<td> *l-A111no-3t5־dichloro-a-[(1nethyla1nino)nethyl]benzyl</td><td> 200</td><td> 15.6</td><td> +56.5</td>

<td> alcohol hydrochloride</td><td> 100</td><td> 18,9</td><td> +87.1</td>

<td> P-amino-a-[(methylamino)methyl|benzyl alcohol hydrochloride</td><td> 100</td><td> IM</td><td> +18.6</td>

<td> a-(9־Wno3,5־-dichlorophenyl)-9-morphollneethanol</td><td> 200</td><td> 15.9</td><td> +36.7</td>

<td> hydrochloride</td><td> 100</td><td> 16.0</td><td> +18.6</td>

<td> *l-flmino-a-|(3ec-butylamino)1Dethyl]’3|5-dichlorobenzyl alcohol ׳</td><td> 200</td><td> 13.2</td><td> +16.8</td>

)

TABLED! (Continued)

Evaluation of Teat Compounds as Animal Growth Promoters

<td> Dosage Gain Conpound (ppm) (grana)</td><td> J Gain Over Controls</td>

<td> 1-taino81־3,5־chloro-&lt;1- |(3־-methoxypropyl)aainol- 200 15.4 methyl benzyl alcohol 100 23.0</td><td> +23.2 +81.0</td>

<td> l-Anino-3,5-dichloro-a-( (dially lam Ino )methyll benzyl 200 1?. 1| alcohol hydrochloride 100 18.4</td><td> +39.2 1 &lt;2 £</td>

<td> 4-Amino-3,5-dichloro-a-l(benzylamino)methyl]benzyl 200 10.6 alcohol hydrochloride</td><td> +19.1 *</td>

<td> 4-Amino-a-[(butylamino)methyl]-3,5-dichlorobenzyl 100 . 14.6 alcohol</td><td> +61.0</td>

<td> 1-Mno-3,5־dlchloro־a4(1-nethyl-l־piperazinyl)- 200 9.1 methyl]benzyl alcohol 100 9.5</td><td> + 5.6 + 6.?</td>

<td> 4-Anino5₎3־-dichloro-a-[(isopropylamlno)methyl]- 200 13.1 benzyl alcohol 100 18.9</td><td> +26.0 +81.7</td>

Z/OSSO9

TABLE (Continued)

Evaluation of Teat Compounds an Animal Growth Pronotera

<td> Compound</td><td> Dosage (ppm)</td><td> Gain (grama)</td><td> $ Gain Over Controls</td>

<td> t-Amino-a-(aminomethyl)-3j5-dichlorobenzyl alcohol hydrochloride</td><td> 100</td><td> 13.4</td><td> +19.0</td>

<td> 4-Amino-3|5-dichloro-a-[(hexylamino)11!ethyljbenzyl</td><td> 200</td><td> 16.8</td><td> +15.9</td>

<td> alcohol</td><td> 100</td><td> 19.2</td><td> +32.8</td>

<td> Q-[(tert־butylamlno)methyl|*3,5*dichlorobenzyl</td><td> 200</td><td> 19.3</td><td> +33.1</td>

<td> alcohol hydrochloride</td><td> 100</td><td> 20.2</td><td> +39.3</td>

<td> A-ta1ino3,5־-dichloro-&lt;1-[(diethylamino)methy1|benzyl alcohol hydrochloride</td><td> 100</td><td> 17.5</td><td> , +20,7</td>

<td> a-KallylaninoJiiiethyll-il-aniino-liS-dichlorobenzyl</td><td> 200</td><td> 16.8</td><td> +118.2</td>

<td> alcohol</td><td> 100</td><td> 17.1</td><td> +122.1</td>

<td>¹l-Amino-a-tanllinoiiiethylJ-JjS-dichlorobenzyl alcohol</td><td> 200</td><td> 20.7</td><td> +25.5</td>

<td></td><td> 100</td><td> 17.5</td><td> + 6.1</td>

TABLE (Continued)

Evaluation Of Teat Compounds as Animal Growth Promoters

<td></td><td> Dosage</td><td> Gain</td><td> f Gain Over</td>

<td> Compound</td><td> (ppm)</td><td> (grams)</td><td> Controls</td>

<td> VAmlno-a-[l-tert-butyla1nlno)ethyl|-3,5-dichloro-</td><td> 200</td><td> 22.7׳</td><td> +37.6</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 23.8</td><td> ♦44.5</td>

<td> i-Amlno-3-bromo-a-[(tert-butylamino)methyl]-5-chloro-</td><td> 200</td><td> 18.5</td><td> +60.9</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 19.5</td><td> +69.6</td>

<td> a-[(tert-butylami.no)methyl|-a-hvdroxvbenzyl alcohol</td><td> 200</td><td> 15.8</td><td> + 9.0</td>

<td> hydrochloride</td><td> 100</td><td> 19.9</td><td> ♦3?. 2</td>

<td> a־[ (isopropy lamino)me thy 1 ] -jn-hydroxybenzy 1 alcohol</td><td> 400</td><td></td><td> +54.4</td>

<td> hydrochloride</td><td> 200</td><td> w</td><td> +53.0</td>

<td></td><td> 100</td><td> t.</td><td> +39.7</td>

<td rowspan="2"> a-[(Amino)methyl|-m-hydroxybenzyl alcohol hydrochloride</td><td> 200</td><td> »</td><td> + 4.4</td>

<td> 100</td><td></td><td> +30.?</td>

<td> 4 - Amino-N-tert-butyl-3,5-dichloro-faiethoxyphenethyl</td><td> 200</td><td> 18.8</td><td> +37.2</td>

<td> hydrochloride</td><td> 50</td><td> 21.6</td><td> +57.7</td>

Z/0ES09

Example

Evaluation of test compounds as animal growth promoters

The procedure of Example 1 is used in this evaluation. The diet is the same as described in said example and data obtained are reported as percent gain over controls. Data are reported in Table IV below.

TABLE IV

Evaluation of Test Compounds a3 Animal Growth Promoters

<td> Dosage</td><td> Initial !Lid</td><td> Final !Lid</td><td> Gain (grams)</td><td> Gain Over Control</td>

<td> 4-Amino-a-(tert-butylaminomethyl)3,5-dichlorobenzyl alcohol hydrochloride</td><td></td><td></td><td></td><td></td>

<td> (ppm in diet)</td><td></td><td></td><td></td><td></td>

<td> 0</td><td> 24.49</td><td> 25.17</td><td> .68</td><td></td>

<td></td><td> 24.25</td><td> 26.06</td><td> 1.81</td><td></td>

<td></td><td> 23.65</td><td> 25.43</td><td> 1.78</td><td></td>

<td></td><td> 22.83</td><td> 24.33</td><td> 1.50</td><td></td>

<td></td><td> 24.39</td><td> 25.59</td><td> 1.20</td><td></td>

<td></td><td> 24.36</td><td> 26.06</td><td> 1.70</td><td></td>

<td></td><td> 23.11</td><td> 24.50</td><td> 1.39</td><td></td>

<td></td><td> 23.54</td><td> 24.82</td><td> 1.28</td><td></td>

<td> Average</td><td> 23.83</td><td> 25.25</td><td> 1.42</td><td></td>

TABLE (Continued)

Evaluation of Teat Compounds as Animal Growth Promoters ί

Gain

<td> P03age</td><td> Initial Mt. (g)</td><td> Final Mt. (g)</td><td> Gain (grama)</td><td> Over Control</td><td> M 0 1</td>

<td> 200</td><td> 23.116</td><td> 25.119</td><td> 2.03</td><td></td><td></td>

<td></td><td> 23.68</td><td> 25.9?</td><td> 2.37</td><td></td><td></td>

<td></td><td> 23.56</td><td> 25.110</td><td> 1.811</td><td></td><td></td>

<td> Average</td><td> 23.5?</td><td> 25.62</td><td> 2.05</td><td> +111.1</td><td></td>

־

Example ______Evaluation of test compounds as animal feed additives for the enhancement of the growth rate of poultry

One day old Hubbard X Hubbard Crossbred Chicks, 5 randomly allotted to pens of ten chicks (5 males and 5 females) each.

Eight pens of chicks are used for unmedicated controls, and four pens of chicks are used at each level of drug. The duration of the experiment is 28 days.

The controls are offered an unmedicated diet of

Broiler Ration No. 453 (composition given below) and water ad libitum. Medicated chicks are offered the same diet containing the test drug at the levels indicated above, and water ad libitum. The weight of the chicks is determined at the beginning and on completion of the experiments. Weight gains and the amount of feed consumed are also determined. The thus obtained data are averaged and summarized in Table V below, wherein the percent improvement in weight gains and feed/gain ratios are given.

<td> 20</td><td> Component</td><td> Percent by Weight</td>

<td></td><td> Ground yellow com</td><td> 53.45</td>

<td></td><td> Soybean oil meal (49%)</td><td> 28.00</td>

<td></td><td> Menhaden fish meal (60%)</td><td> 5.0</td>

<td></td><td> Corn gluten meal (60%)</td><td> 5.00</td>

<td> 25</td><td> Dehydrated alfalfa meal (17%)</td><td> 2.00</td>

<td></td><td> Stabilized fat</td><td> 4.00</td>

<td></td><td> Dicalcium phosphate</td><td> 1.20</td>

<td></td><td> Ground limestone</td><td> 0.50</td>

<td></td><td> Sodium chloride</td><td> 0.30</td>

<td> 30</td><td> Trace minerals mixture*</td><td> 0.05</td>

<td></td><td> Vitamin premix*♦</td><td> 0.50</td>

<td></td><td></td><td> 100.00</td>

<td></td><td> *Trace Mineral Mixture________</td><td> 1 Ib/ton furnishes</td>

<td> 35</td><td> Manganese 12.50 %</td><td> 62.5 ppm</td>

<td></td><td> Iron 6.00</td><td> 30.0</td>

<td></td><td> Zinc 5.00</td><td> 25.0</td>

<td></td><td> Copper 0.65</td><td> 3.25</td>

6Θ530/2

Iodine 0.351.75

Cobalt 0.251.25

Calcium minimum15.30

Calcium maximum17.35 **Vitamin Premix for 1-ton Weight in Gram

DL Methionine453.6

BHT (butylated hydroxy toluene)113.6

Vitamin A (30,000 mcg/g)100.0

Vitamin Dj (200,000 mcg/g)5.0

Vitamin E (20,000 mcg/lb)45.4

Riboflavin4.0

Niacinamide25.0

TABLE V,

Kean Weight Gain and Feed Efficiency of Control and Test Compound * Treated Chicks

Thirteen-Day Battery Testing

<td></td><td> ppm</td><td> Mean</td><td> ן</td><td></td><td> ί Improvement</td>

<td> Treatment</td><td> in Diet</td><td> Gain (g)</td><td> Control</td><td> F/G</td><td> over Control</td>

<td> Control</td><td> 0</td><td> 266.5</td><td> «</td><td> 1.90</td><td></td>

<td> l|-Amino״a-| (tert-butylamlno)methyll</td><td> 0.3</td><td> 271.8</td><td> +2.0</td><td> 1.90</td><td> 0</td>

<td> 3,5-dichlorobenzyl alcohol</td><td> 0.6</td><td> 2713</td><td> ♦2.9</td><td> 1.38</td><td> +1.¢</td>

<td></td><td> 1.25</td><td> 259.9</td><td> 5.²־ .</td><td> 1.91</td><td>.7</td>

<td></td><td> 2.5</td><td> 260.8</td><td>.1</td><td> 1.39</td><td> +0.7</td>

<td></td><td> 5.0</td><td> 251.9</td><td>.5</td><td> 1.90</td><td> 0</td>

<td> i-Amino-3,5-dibromo-a-[(tert-butyl-</td><td> 0.3</td><td> 270.8</td><td> +1.6</td><td> 1.90</td><td> 0</td>

<td> aminojmethyl)benzyl alcohol hydrochloride</td><td> 0.6</td><td> 271.8</td><td> +2.0</td><td> 1.38</td><td> +1.¢</td>

<td></td><td> 1.25</td><td> 267.5</td><td> +0,9</td><td> 1.39</td><td> +0.7</td>

<td></td><td> 2.5</td><td> 265.2</td><td>.5</td><td> 1.38</td><td> +1.¢</td>

<td></td><td> 5.0</td><td> 270.3</td><td> +1.9</td><td> 1.37</td><td> +2.1</td>

24 ־

Example 4 Evaluation of test compounds as antilipogenic agents - . Mouse tests

CFI female mice from Carworth Farms are received when they are six weeks old. They are housed ten to a cage in air-conditioned rooms (72°F to 76°F) with automatically controlled lights, 14 hours on and 10 hours off. The basal diet used in these studies is Purina Laboratory Chow (see description below), which is supplied ad libitum.

The following is a description of the diet to which the growth-promoting compounds were added.

DIET

Guaranteed Analysis

<td> Crude protein not less than</td><td> 23.0%</td>

<td> Crude fat not less than</td><td> 4.5%</td>

<td> Crude fiber not more than</td><td> 6.0%</td>

<td> Ash' not more than</td><td> 9.0%</td>

<td> Ingredients</td><td></td>

Meat and bone meal, dried skimmed milk, wheat germ meal, fish meal, animal liver meal, dried beet pulp, ground extruded corn, ground oat groats, soybean meal, dehydrated alfalfa meal, cane molasses, animal fat preserved with BHA, vitamin supplement, calcium pantothenate, choline chloride, folic acid, riboflavin supplement, brewer's dried yeast, thiamin, niacin, vitamin A supplement, D-activated plant sterol, vitamin E supplement, calcium carbonate, dicalcium phosphate, iodized salt, ferric ammonium citrate, iron oxide, manganous oxide, cobalt carbonate, copper oxide, zinc oxide. Water is also allowed ad libitum.

Thirteen days after arrival, the mice are weighed in groups of ten and assigned at random to the different treatments. The concentration of the different compounds in the diet is indicated in the following tables. Twelve days later the mice are weighed again the experiment terminated. At least three cages (30 mice) of untreated controls are included in each test. Test data are provided in Table VI below wherein data are reported, as percent body fat, percent change in body fat from controls and gain per mouse

TABLE

Antilipogenic Agent Evaluation and Growth Enhancement Evaluation In Mice

<td></td><td> Level</td><td> Number</td><td> Average</td><td> Average</td><td> Gain</td><td></td><td> Change</td>

<td></td><td> in</td><td> of Mice</td><td> Initial</td><td> Final</td><td> per</td><td> J</td><td> in$</td>

<td></td><td> Diet</td><td> per</td><td> Weight</td><td> Height</td><td> House</td><td> Body</td><td> Fat from</td>

<td> Compound</td><td> (ppm)</td><td> Treatment</td><td> JiL</td><td> (g) .</td><td> (g)</td><td> Fat</td><td> Control</td>

<td> 01</td><td> 0</td><td> 50</td><td> 23.6</td><td> 25.0</td><td> 1.1</td><td> 11.95</td><td></td>

<td> / Λ 1.H-/ ;.M,־m(CH,),</td><td> 50</td><td> 30</td><td> 23.7</td><td> 26.0</td><td> 2.3</td><td> 11.95</td><td> 0</td>

<td> 2 \ / I J J .=.</td><td> 100</td><td> 30</td><td> 23.1</td><td> 25.7</td><td> 2.3</td><td> 10.23</td><td> : -11.10</td>

<td> ii</td><td> 200</td><td> 30</td><td> 23.5</td><td> 25.3</td><td> 1.8</td><td> 10.50</td><td>,13</td>

<td></td><td> 100</td><td> 30</td><td> 23.1</td><td> 21.9</td><td> 1.1</td><td> 9.10</td><td>.80</td>

&lt;/βίίίΚ»»

Percent Body Fat Determination of Mice

A. Preparation of Carcasses:

Stomach and intestines are removed from each mouse. All other viscera, including skin and fur, remain intact. Each cage of mice (10) are weighed and added to a 1000 ml beaker and autoclaved at 120°C (15 psi) for 30 minutes. Carcasses from each cage are then blended and homogenized. The homogenate is weighed and duplicate 5-gram samples are removed for analysis.

B. Fat Analysis:

Fifteen milliliters (ml) of concentrated hydrochloric acid is added to each 5-gram sample and mixed well. Samples are heated in an 84°C water bath for 2 hours. To extract the fat, thirty ml of petroleum ether is added to each samples, 15 ml at a time, and mixed well on a Vortex mixer. The aqueous and organic phases are separated by low speed centrifugation and the ether layer (containing fat) is extracted into tared 30 ml beakers. After evaporating to dryness the beaker containing fat is reweighed to determine grams of fat per five grams of homogenate. Total body fat in the carcass is calculated as follows:

<td> % Fat =-</td><td> grams fat in sample</td><td></td><td> grams total homogenate</td><td></td>

<td></td><td> gram weight' of sample</td><td></td><td colspan="2"> ------------------ X 1UU carcass weight of mice (g)</td>

Example

Antilipogenic Evaluation of test compounds - Mouse Study

CFI female mice, 55 days old, are weighed in groups of 10 and allotted to cages to minimize weight variation among cages. Treatments are randomly assigned to cages.

Each of the treatments are tested in 3 replicates,

i.e., in 3 cages of 10 mice each. There are 10 cages of 10 control mice each. Durgs are mixed in the diet at the dosage level indicated. Feed and water are offered ad libitum for the 12-day test period. Feed spilled is collected during the test period. At the end of the test period, the collected feed is weighed and the mean feed consumption per cage of ten mice is determined for each treatment. The mice are. weighed as a group of 10 and the weight gain determined. The miceare sacrificed by cervical dislocation. The right uterine fat pad of each mouse is removed. The fat pads for each cage of 10 mice are weighed as a unit.

To establish correlation between the percent reduction in fat pad weights of treated animals and percent reduction in total body fat of treated animals, animals from several treatment groups are evaluated for total body fat using the body fat determination described in Example 5.

Data obtained are reported in Table VII for those groups upon which such determination has been made. From percent reduction in fat pad weight and the total fat determinetions for the groups tested, it can be seen that a reduction 15 in fat pad weights of animals is generally indicative of a reduction of total body fat of the treated animals.

<img file="IL60530A_D0007.tif" />

TABLE

Antillpogenic Evaluation of Test Compounds - Mouse Study

<td> / י Compound</td><td> Dosage (PPH) k</td><td> $ Reduction in Fat Pad Weight vs Controls</td><td> J Animal Fat vs Controls</td>

<td>¹l-amino-3,5-dibromo״a-[ (tert-butylamino)methyH benzyl</td><td> wo</td><td>.4</td><td> 14.6</td>

<td> alcohol hydrochloride</td><td> 200</td><td> • 27.5.</td><td>.4.</td>

<td></td><td> 100</td><td> 13.9.</td><td>.3'</td>

<td> 4-amino-3,5-dibromo-a-[(diisopropylamino)methyl|benzyl alcohol hydrochloride</td><td> 200</td><td>.1</td><td>.4</td>

<td> ^amino-a-[(tert-butylamino)methyl]-3.5־dichlorobenzyl</td><td> 400</td><td> 50.0</td><td>.8</td>

<td> alcohol hydrochloride</td><td> 200</td><td>.1</td><td> 12.1</td>

<td></td><td> 100</td><td> •37.9</td><td>.4</td>

<td> Camino-3&gt;5-dichloro-a-((methylamino)methyl|benzyl</td><td> 200</td><td>.7</td><td>.9</td>

<td> alcohol hydrochloride</td><td> 100</td><td> 8.8</td><td>.5</td>

<td>¹l-amino-J^-dichloro-a-[ (diethyl)methyl]benzyl alcohol</td><td> 200</td><td>,7</td><td></td>

<td> hydrochloride</td><td> 100</td><td>.2</td><td></td>

<td>⁴l-amino-a-l (sec-butylamino)methyll5|3־-dichlorobenzyl</td><td> 200</td><td>.2</td><td> 36.3־</td>

<td> alcohol</td><td> 100</td><td>.2</td><td>.3</td>

¹Average 2 tests

TABLEVI (Continued)

Antilipogenlc Evaluation of Teat Compounda Mouse Study

<td> Compound</td><td> Dosage (ma)</td><td> J Reduction in Fat Pad Height vs Controls</td><td> J Animal Fat vs Controls</td>

<td>¹l-anino-SfS-dichloro-a- ((dlallylamino)methyl] benzyl alcohol hydrochloride</td><td> 200</td><td> . -12.0</td><td></td>

<td> if-aaino-J^-dlchloro-a-Ifbenzylaiiinoliiethyllbenzyl</td><td> 200</td><td>.7</td><td> י.5.</td>

<td> alcohol hydrochloride</td><td> 100</td><td>.1</td><td>.7</td>

<td> ll-amino-a-[(butylamino)methyl]-3,5-dichlorobenzyl</td><td> 200</td><td>.54</td><td>.78</td>

<td> alcohol</td><td> 100</td><td>.7</td><td>.08</td>

<td>¹l-amino-JjS-dichloro-a-ldsopropylaniinolmethyllbenzyl</td><td> 200</td><td>.2</td><td> •25.5</td>

<td> alcohol</td><td> 100</td><td> •36.9</td><td>.4</td>

<td> a-[(allylamino)methyl]-i-amino-jiS-dichlorobenzyl</td><td> 200</td><td>.5</td><td></td>

<td> alcohol</td><td> 100</td><td>.3</td><td></td>

<td> i-amino-a-[l-(tert-butylamino)ethyl]-3j5״dichlorobenzyl alcohol hydrochloride</td><td> 100</td><td>.8</td><td></td>

<td> a-[(t-butylamino)methyl]-3,5-dichlorobenzyl</td><td> 200</td><td>.5 .</td><td></td>

<td> alcohol hydrochloride</td><td> 100</td><td>.8</td><td></td>

™YL !continued)

Antilipogenlc Evaluation of Test Compounds - Mouse Study

I Reduction in Pat

<td> Compound</td><td> Dosage (ppm)</td><td> Pad Weight vs Controls</td><td> I Animal Fat vs Controls</td>

<td> 4-amino-3-bromo-a-} (t-buty lamino) me thy 1J 5־-chloro</td><td> 200</td><td>.8</td><td></td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 18.7־</td><td></td>

<td> rhydroxy-a-J(isopropylamino)methyljbenzyl alcohol</td><td> 400</td><td>,8</td><td></td>

<td></td><td> 200</td><td>.2</td><td></td>

<td></td><td> 100</td><td>.5</td><td></td>

<td> 4-amino-N-t-butyl-3j5-dichlorophenethylamine hydrochloride</td><td> 50</td><td>.8</td><td></td>

<td> 4-amino-3,5-dichlor0-a- I(cyclopropylamino)methyljbenzyl alcohol</td><td> 100</td><td>.7</td><td></td>

<td> 4-{2-(t-butylamino)-l-hydroxyethyl]-2^l-chloroacetanilide</td><td> 200</td><td>.7</td><td></td>

<td></td><td> 100</td><td>.1</td><td></td>

<td> 4-amino-3 ^״dichloro-a-I (cyclopentylamino)methyl]benzyl</td><td> 200</td><td> 24,5־</td><td></td>

<td> alcohol</td><td> 50</td><td> 4.7־</td><td></td>

<td> 4-amino-3,5-dichloro-a-{] [2-hydroxyethyl)amino]methyl}-</td><td> 200</td><td>.2</td><td></td>

<td> benzyl alcohol</td><td> 50</td><td>.4</td><td></td>

<td> 4-amino-a-[(t-butylamino)methyl]-3,5-diiodobenzyl alcohol</td><td> 200</td><td>.6</td><td></td>

<td> hydrochloride</td><td> 100</td><td>.6</td><td></td>

<td> 4-amino-N-t-butyl-3,5-dichloro-Hethoxyphenethylamine</td><td> 200</td><td>.4</td><td></td>

<td> hydrochloride</td><td> 50</td><td> 21.7־</td><td></td>

Example 6

Antilipogenic evaluation of test compounds - Rat study The procedure employed and the diet used for evaluation of test compounds as antilopogenic agents mice, 5 are described in Example 1, excepting that the treatment period is fourteen days and 10 rats, one per cage, are used for each treatment.

Percent body fat is determined in the same manner as described in Example 4, excepting that the skin and organs 10 are removed before the carcasses are homogenized.

Results of this study are reported in Table VIII below.

TABLE VII

Antilipogenic Evaluation of Test Compounds and Growth Rate Evaluation in Rats

Compound

ף /^,A ?^{H h}2^h\ .=. ci

<td> Level</td><td> Number</td><td> Average</td><td> Average</td><td colspan="2"> Average</td><td> Change</td>

<td> in</td><td> of Rats</td><td> Initial</td><td> Final</td><td> Gain</td><td> J Fat in</td><td> in Fat</td>

<td> Diet</td><td> per</td><td> Weight</td><td> Height</td><td> per Rat</td><td> Eviscerated</td><td> from</td>

<td> (ppm)</td><td> Treatment</td><td> JsL</td><td> (g)</td><td> JfiL</td><td> Carcass</td><td> Control</td>

<td> 0</td><td> 10</td><td> 72.7</td><td> ' 149.1</td><td> 76.11</td><td> 1.67</td><td> יי M</td>

<td> 25</td><td> 10</td><td> 78.3</td><td> 159.3</td><td> 81.0</td><td> 3.04</td><td>.9</td>

<td> 100</td><td> 9</td><td> 76.6</td><td> 159.8</td><td> 83.2</td><td> 2.5*1</td><td> 12.0</td>

<td> ¢00</td><td> 10</td><td> 73.2</td><td> 146.4</td><td> 73.2</td><td> 2.71</td><td> -*15.6</td>

³³ 60SW2

EXAMPLE Z ר

Evaluation of test compounds as animal feed additives for the enhancement of growth rate and improvement in feed efficiency of mice.

Four-week old female outbred rats (5-gram range) from Charles River Breeding Laboratories, 251 Ballardvale Street, Wilmington, Massachusetts 01887, are housed 2/cage in air-conditioned rooms (72^eF to 76°F) with automatically controlled lights, 14 hours on and 10 hours off. The basal 10 diet used in these studies is Purina Laboratory Chow which is supplied ad libitum. Water is also given ad libitum.

Four days after arrival, the animals are weighed and allotted to treatment groups to minimize weight variation. Ten rats are used per treatment group. Drugs are 15 administered in the feed at 2 ppm, 10 ppm and 50 ppm for a period of 12.5 weeks. Animals are weighed weekly and feed consumption corrected for spillage recorded daily. The results of this trial are shown below in Table IX.

TABLE

Evaluation of Test Compounds as Animal Feed Additivea for the Enhancement of Crowth Bate and Improvement In Feed Efficiency - Mice k Feed .

Dose Gain⁸ Consumption Feed/Galn

<td> Treatment</td><td></td><td></td><td> ___ill—</td><td> ( Improvement</td>

<td> Control</td><td></td><td> 157</td><td> 1304</td><td> 8.31</td>

<td> l|-Amino״a־(tert־butylaminomethyl)-3,5־dlchloro-</td><td> 2</td><td> 178 (+13.13)</td><td> 1443 (+10.73)</td><td> 8.11 (+2.1()</td>

<td> benzyl alcohol hydrochloride</td><td> 10</td><td> 186 (+18.53)</td><td> 1167 (+11.53)</td><td> 7.89 (+5.1()</td>

<td></td><td> 50</td><td> 175 (+11.53)</td><td> 1394 (+6.9()</td><td> 7.97 (+1.1()</td>

<td> 1-taino-3,5-dlbrono^-[(tert-butylaBino)11ethyll.</td><td> 2</td><td> 164 (+1.5()</td><td> 1362 (+1.5()</td><td> 8.3 (0.13)</td>

<td> benzyl alcohol hydrochloride</td><td> 10</td><td> 185 (+7.83)</td><td> 1459 (+11.9()</td><td> 7.89 (+5.1()</td>

<td></td><td> 50</td><td> 184 (+17.23)</td><td> 1416 (+8.6()</td><td> 7.70 (+7.3()</td>

®

Values given are the total average gain (g) per rat for the entire experimental period, 5 ^Values given are the total average feed consumed per rat for the entire experimental period. w Figures in parentheses are J improvement over control,

EXAMPLE 8 a-[(Tert-butylamino)methyl] -3,5-dichlorobenzy1 Alcohol Hydrochloride

A solution containing 3.5 g of 3,5-dichlorostyrene 5 oxide in 50 ml of absolute ethanol and 20 ml of t-butylamine is heated gently at reflux for 8 hours and the mixture is evaporated to dryness. The clear yellow syrup is dissolved in 75 ml of ethanol and 25 ml of H₂O, and the solution is cooled to.5״C and acidified with 3N HC1. This 10 solution is evaporated to dryness in vacuo and the residual white solid is recr-stallized from acetone to afford . 2.81 g, m.p. 218-221°C.

Anal . Called for C,-H.-NOC1-HC1: C, 48.26; H, 6.08; H, 4.69. X Μ X / M ^Λ Found: C, 48.49; H, 6.17; N, 4.66. 15 The free base of the title compound is obtained > by neutralization of the title compound with aqueous 10% b NaOH. Other salts of the free base are then obtained by treatment of the free base in the above-mentioned procedure (aqueous ethanol) with addition of the appropriate acids, 20 such as HjSO^, H^PO^, HNO^, CH^SO^H, toluenesulfonic acid and pantoic acid.

The intermediate 3,5-dichlorostyrene oxide needed for preparing the title compound is made by reducing 28.4g of 3,5-dichlorophenacyl bromide in 125 ml of absolute ethanol 25 at 5°C with 8g of NaBH₄, added portionwise. After the addition is completed, the reaction mixture is stirred 16 hours at ambient temperature, which is obtained by gradual melting of the ice bath overnight. The mixture is quenched with 100 ml of H₂O, the aqueous mixture is cooled to 5°C, 30 and carefully acidified to pH 3 with concentrated HC1. The mixture is extracted with 300 ml of CH₂C1₂ and the extract is dried over MgSO₄, filtered, and evaporated to dryness in vacuo to afford the epoxide as a clear yellow oil.

The phenacyl bromide intermediate for the above35 mentioned styrene oxide is prepared by brominating 10 g of

3,5-dichloroacfetophenone in 50 ml of CHCl^/SO ml of EtOAc with 23.6 g of CuBr₂״ The mixture is heated at reflux for

2.5 hours and cooled to room temperature. After stirring for 16 hours at room temperature, the mixture is cooled in ice for 2 hours and filtered. The filter cake is washed with 50 ml of CHCl^ and the combined filtrates are twice decolorized with activated carbon, filtered, and evaporated to dryness in vacuo to afford the orange oil of the 3,5dichlorostyrene oxide.

EXAMPLE ⁹

The following 3,5-dichlorophenyl compounds (A) related to the title compound of Example 8 are prepared by the method described in Example 6 by substituting t-butyl amine with R2^r3ⁿH amines.

Ji Or™ ii

A

Compound ^R2

Ηh ² HCH ^{3 H}C״H.

it V ^{4 H} &lt;־c₃h₇^{5 H} S־C₃^H7 ^{6 H} n-C_fiH..

ם ״ H cyclohexyl ⁸ H CH₂-CH=CH₂^{9 H} CH₂-CH=CH-CH₃¹⁰ H CH₂C=CH

A (Continued)

<td></td><td> Compound</td><td></td><td> ב</td>

<td></td><td> 11</td><td> H</td><td> phenyl</td>

<td> כ</td><td> 12</td><td> H</td><td> methoxypropyl</td>

<td></td><td> 13</td><td> H</td><td> benzyl</td>

<td></td><td> 14</td><td> ch₃</td><td> CH״ &lt;5</td>

<td></td><td> 15</td><td>^C2^H5</td><td>^C2^HS</td>

<td> 10</td><td> 16</td><td> ch₂ch־ch₂</td><td> CH₂CH=CH₂</td>

<td></td><td> 17</td><td> i-c₃K₇</td><td> ί-ΟβΗγ</td>

<td></td><td> 18</td><td> ch₂-ch־ch₂</td><td> -CH₂-CH=CH₂</td>

<td></td><td> 19</td><td> H</td><td> cyclopropyl</td>

<td></td><td> 20</td><td colspan="2"> -CH₂-CH₂-O-CH₂-CH₂-</td>

<td> 15</td><td> 21</td><td> H</td><td> n-butyl</td>

<td></td><td></td><td></td><td></td>

<td></td><td> 22</td><td> H</td><td> C(CH₃)₂-CH₂\ ;.</td>

<td></td><td></td><td></td><td></td>

<td></td><td></td><td> EXAMPLE</td><td> • 10</td>

<td></td><td colspan="2"> a-[(Tert-butylamino)methyl]-3,5-</td><td> dibromobenzyl Alcohol Hydro-</td>

<td> 20</td><td> chloride</td><td></td><td></td>

This title compound is prepared from 3,5-dibromostyrene oxide in the same manner as described in Example 9. The starting materials for this styrene oxide are similarly prepared starting with 3',5’-dibromoacetophenone.

The corresponding a-[(isopropylamino)methyl]-3,5dibromobenzyl alcohol hydrochloride is prepared by substituting isopropyl amine for t-butyl amine.

EXAMPLE 11 m-Hydroxy-a-[(isopropylamino)methyl]benzyl Alcohol Hydrochlo30 ride

In 135 ml of 95% ethanol, 36.75 g of m-hydroxyacetophenone, 36.5 g of benzyl chloride, 1.75 g of KI, and 24.6 g of KjCOj are stirred and heated at reflux for 5 hours. The mixture is cooled, evaporated in vacuo to remove ethanol and 35 100 ml of HjO is added. The mixture is then extracted with diethyl ether three times to afford 350 ml of extract, which is further washed with 50 ml of H₂O, saturated NaHCO^ solu tion (2 x 50 ml), 50 ml of H₂O, and 50 ml of brine in succession. The filtrate is dried over Na₂SO₄ and evaporated to dryness. The residual oil is distilled to afford 49.13 g of m-benzyloxyacetophenone, b.p. 145147°־C/0.2 mm. Bromination of 186 g of this acetophenone is accomoplished with 349 g of CuBr₂ in 1£ of CHCL₃/1.5 A of ethanol heated at reflux. A N₂ sweep is used to remove HBr generated. After 4 hours, the mixture is filtered and the filter cake is washed with CHC1₃ (2 x 100 ml)« The filtrate is evaporated in vacuo to afford an oil, which is dissolved in 200 ml of absolute ethanol (2 x 50 ml), and dried to afford 64.28g m-benzyloxyphenacyl bromide, m.p. 5758־*C. Further cooling of the filtrate affords 34g. A 64 g-sample of the phenacyl bromide is added to a stirred mixture containing 212 ml of i-propyl15 amine in 425 ml of ethanol under N₂ atmosphere at 5°C. The temperature rises to 12°C and a clear solution is obtained. The solution is poured into ice (2 L) containing 500 ml of concentrated HC1 and 1500 ml of H₂0. After stirring for 20 minutes, the mixture is filtered and the solid is washed with H₂O. On drying this gives 98.64g, m.p. 200203°־C dec. This solid is dissolved in 400 ml of refluxing methanol, 400 ml of isopropyl alcohol is added, and the solution is concentrated to 400 nl. On cooling and collecting crystals, 54.36 of ketoamine melting at 213215°־ dec is obtained. This material (16 g) is added to 150 ml of methanol which contains 2 g of 5i Pd/carbon and hydrogenated in a Paar vessel at 42 p.s.i.g. of H₂. The mixture is filtered and the filtrate is evaporated. The residue is mixed with 50 ml of isopropyl alcohol and evaporated to dryness to afford a syrup, which is mixed with 100 ml of ethanol. They crystals are collected, washed with diethyl ether and dried to give 10.77 g, m.p. 129-132°C, of the title compound.

By substituting tert־butylamine for isopropylamine, m-hydroxy־a-[(tert-butylamino)methyl]benzyl alcohol hydrochloride, m.p. 150154־*C dec. is obtained. Substitution of isopropylamine with diisopropylamine, benzylamine and allyamine affords m־hydroxy־a־[(diisopropylamino)methyl] benzyl alcohol, m-hydroxy-a-[(benzylamino)methyl]benzyl alcohol, and m-hydroxy-a-[(allylamino)methyl]benzyl alcohol hydrochlorides, respectively.

EXAMPLE 12

4-Amino-a [ (tert-butylamino)methyl]-3,5-diiodobenzyl Alcohol HxtochlflKide

In 10 ml of acetic acid, 0.42 g of g-amino-a[(tert-butylamino)methyl]benzyl alcohol is stirred under N₂atmosphere and 0.48g of Ν,Ν-dichlorobenzenesulfonamide and

0.6g of Nal are stirred under N₂ atmosphere for 20 minutes. After 3 days, the mixture is poured into ice and the mixture is basified with 50% aq. NaOH. This mixture is extracted with CH₂C1₂ (3 x 25 ml) and chromatographed on a SiO₂ plate using 1% NH₄OH/20% CH₃OH/CH₂C1₂ to afford 0.22g of the title 15 compound. The reaction is repeated on a larger scale (8X) and the eluted crude product is dissolved in 100 ml of ethanol/10 ml of HjO, stirred and 10% HC1 is added to give pH 3. The mixture is evaporated to dryness in. vacuo. Isopropyl alcohol is added and the mixture is evaporated to dryness. This process is repeated twice and the residue is crystallized from methanol/isopropy1 alcohol by allowing methanol to evaporate until crystals from (methanol is used to dissolve the crude material before isopropyl alcohol is added). On cooling, 2g of the title compound is obtained melting at 187°C dec.

Anal. Calc'd for C₁₂H₁₉C1I₂N₂O: C,29.02; H,2.86; N,5.64.

Found:. C, 29.11; H, 3.64; N, 5.64.

EXAMPLE .13 a-((Tert-butylamino)methyl]-3,5-dichlorobenzyl Alcohol Hydro30 chloride

An alternate procedure for preparing the title compound and the compounds described in Example is exemplified. Thus, 10 g of 4-amino-a-[(tert-butylamino)methyl]-3,5dichlorobenzyl alcohol is added to 100 ml of 50-52% H₃PO₂35 and the mixture is stirred and cooled to 8°C in ice while 2.77 g of NaNO₂ in 15 ml of HjO is added over 65 minutes. Foaming occurs and is controlled with antifoaming silicone.

6ΟΠ

After 20 minutes, the mixture is stirred 2 hours without cooling. The mixture is then poured into ice-HjO mixture and 50% aq. NaOH solution־is added until the mixture is alkaline. The alkaline mixture is extracted with CHjClj three 5 times to give 200 ml of ^,solution, which is washed with 25 ml of 2% NaOh and dried over MgSO^ and evaporated to dryness in vacuo to give 9.13 g of brown oil. ^n standing, the oil solidifies, and it is dissolved in 100 mB of ethanol containing 10 ml of The solution is acidified to pH 3 with 10 10% HC1 and evaporated to dryness. The residue is treated with 50 ml of isopropyl alcohol and evaporated to dryness.

This procedure is repeated to afford an off-white solid which is. dissolved in methanol. The solution is evaporated in vacuo to afford a syrup, which is diluted with 50 ml of iso15 propyl alcohol and allowed to stand. The crystals which form are collected, washed with isopropyl alcohol and dried to yield 7.8 g, m.p. 217-221®C dec., of the title compound.

The compound described in Example 6 is similarly prepared. Deamination of 4-amino-3,5-dibromo-a-[(tert-butyl20 amino)methyl]benzyl alcohol affords 3,5-dibromo-a-[(tertbutylamino)methyl]benzyl alcohol, m.p. 249-251’C dec.

EXAMPLE ' 14

4-Amino-3,5-dichloro-8-methoxyphenethylamine hydrochloride Under atmosphere, 11 g of 4-amino-a-[(tert-butyl25 amino)methyl]-3,5-dichlorobenzyl chloride is added to 75 ml of methanol at 0״C. After 20 minutes, the cooling bath is removed and the reaction mixture is stirred at ambient temperature. After the reaction is completed, the mixture is evaporated to dryness in vacuo. The residue is stirred in 30 75 ml of H₂0 and the mixture is made alkaline with 6N NaOH solution and extracted with CH₂C1₂ (3 x 50 ml). The organic phases are dried over MgSO₄ and evaporated to dryness to afford an orange oil. This oil’is dissolved in 150 ml of absolute EtOH and acidified with HCl/isopropyl alcohol solu35 tion to pH 2. The solution is evaporated to dryness and the residue is stirred in 75 ml of ethyl acetate. After cooling, this affords a pale yellow solid which is 'collected to give

6.97 g of the title compound, m.p. 195-198^OC dec.

Similarly, substitution of ethyl alcohol, isopropyl alcohol, n-butyl alcohol and n-hexyl alcohol affords the corresponding B-ethoxy, β-isopropoxy, n-butoxy, and n-hexyl5 oxy phenethylamine hydrochlorides.

EXAMPLE 15

4-Amino-a-[(tert-butylamino)methyl]-3,5-dichlorobenzyl chloride

Under N₂ atmosphere, 27.72 g of 4-amino-a-[(tert10 butylamino)methyl]-3,5-dichlorobenzyl alcohol is added to 200 ml of thionyl chloride stirred at 0-5%C. After addition is completed, the reaction mixture is stirred at ambient temperature for 3 hours. Subsequently, the mixture is evaporated to dryness in vacuo to afford 37.34 g of yellow solid, 15 which is used as is.

EXAMPLE . . 16

Alternate Procedure for 4-Amino-3,5-dichloro-B-methoxyphenethylamine

I. 100 ml of methanol, 10 g of 4-amino-a-[(tert20 butylamino)methyl]-3,5-dichlorobenzyl alcohol is stirred in an ice bath and dry HC1 gas is introduced into the ,solution. After saturation of the solution, the mixture is stirred at toom temperature for an hour and evaporated to dryness. The solid is then stirred in ethyl acetate to afford the title product, which is collected by filtration.

ANIMAL FEED COMPOSITIONS USEFUL AS GROWTH PROMOTERS AND FOR REDUCTION OF FAT IN ANIMALS, COMPRISING PHENYLETHANOLAMINE DERIVATIVES AND CERTAIN SUCH NOVEL COMPOUNDS

תכשירי מזרן לבעלי חיים המשמשים כמעודדי גדילה ולהפחתת שומו בבעלי חיים

המכילים תולדות פנילאתאנולאמיו ותכשירים חדשים מסויימים כאלה / ־^;׳ Substitution products of certain 1- (aminodihalo.'־ / j phenyl)-2-aminoethanes and the acid addition salts thereof ’ y/' are disclosed in United States Patent 3,536,712, (corresponding to local patent No,28662) issued on 27th October, 1970.

' Said Israeli Patent No. 28662 discloses similar ^r compounds which are useful for enhancing the blood circulation, and as bronchodilators, analgesics, sedatives, antipyretics, antiphlogistics and antitussive which is entirely different from the utility f of the compounds of this invention.

The U. S. Patent 3,818,101 discloses the use of certain -adrenergic agents for improving the intake of feed in meat-producing animals, whereas the compounds ‘׳‘'־ ^r of the instant application generally depresses feed intake.

Specifically, patentees disclose methods for the synthesis of said compounds and state that said compounds are useful for enhancing the blood circulation, and as bronchodilators, analgesies, sedatives, antipyretics, antiphiogistics and antitussives in warm-blooded animals. Patentees, however, exemplify only the analgesic utility. They do not indicate or suggest that said compounds are useful for lowering the deposition of fat or increasing the growth rate in worm-blooded animals, particularly fax־m and domestic animals, such as swine, poultry, dogs, sheep, goats, cats or cattle.

It has now been found that the growth rate and the depression of fat deposition of meat-producing animals such as swine, chickens, turkeys, domestic pets, rabbits, sheep, goats and cattle, including calves, can be increased and the efficiency of feed utilization thereby measurably improved by the oral or parenteral administration to said animals of an effective amount of a compound having the structure: X

Y—/^/'J\־ch-ch-nr₂r₃

Z ^1 wherein X is hydrogen or halogen ( chlorine or bromine) ; Y is hydrogen,

9.7.80 NH₂ or NHCOR₅; Z is H, halogen ( chlorine or bromine) or OH; R^ is hydrogen] R₂ ^-³ hydrogen, C₁~C₄ alkyl (straight or branched-chain) or alkenyl; Rg is hydrogen, C^-Cg alkyl (straight or branched-chain), Cg-Cg cycloalkyl, methoxypropyl, C^-C^ alkenyl, phenyl, 2-hydroxyethyl, a-a-dimethylphenethyl or benzyl;

are taken together with the nitrogen to and when R₂ and Rg which they are

9.7.80

9.7.80 attached, they may represent morpholino or N'-C^-C₄ alkylpiperazine; R₄ is hydrogen, hydroxyl or OR₆; R₅ is

Cj-C^ alkyl; Rg is C-^-Cg alkyl; with the provisos that when Rg ^³ phenyl, 2-hydroxyethyl, a,a-dimethylphenethyl, cycloalkyl Cg-Cg, benzyl or methoxypropyl, R₂ is hydrogen; and when Z is OH, X and Y are hydrogen; and when Y is NHCORg, at least one of X and Z is hydrogen; and provided also that at least one of X,Y and Z represents a substituent other than hydrogen; racemic mixtures of the above-identified compounds and the optically active isomers and non-toxic, pharmacologically acceptable acid addition salts thereof.

Preferred compounds for use in the method of this 20 invention have the above structure wherein X and Z are each chlorine or bromine; Y is hydrogen or NH₂; R-j is hydrogen or C₁״C₄ alkyl; R₄ is hydroxyl; or a. non-toxic, pharmacologically acceptable acid addition salt thereof. Some of the said compounds are novel and are claimed as such.

The most preferred compounds for use in enhancing the growth rate of meat-producing animals and for improving the efficiency of feed utilization thereby are: 4-amino-a[(tert-butylamino)methyl]-3,5-dichlorobenzyl alcohol hydrochloride; 4-amino-3,5-dibromo-a-[(diisopropylamino)methyl]benzyl alcohol hydrochloride; 4-amino-3,5-dichloro-a-[(di30 isopropylamino)methyl]benzyl alcohol hydrochloride; 4amino-3,5-dibromo-a-((tert-butylamino)methyl]-benzyl alcohol hydrochloride; 4-amino-3,5-dichloro-a-[(methylamino)methyl]benzyl alcohol hydrochloride; 4-amino-3,5-dichloro-a-[(allylamino)methyl]benzyl alcohol; 4-amino-3-bromo-a-[(tert35 butylamino)methyl]-5-chlorobenzyl alcohol hydrochloride;

a-[4-amino-3,5-dichlorophenyl]-4-morpholineethanol hydrochloride; 4-amino-3-bromo-a-[(tert-butylamino)methyl]-5-360530/2 chlorobenzyl alcohol hydrochloride and a-[(tert-butylamino) methyl]-3,5-dichlorobenzyl alcohol hydrochloride.

It is found, that formula (I) compounds below (wherein Y is hydrogen) can be prepared by the condensation of an appropriately substituted sytrene oxide with the appropriately substituted amine in the presence of an inert solvent, such as a lower alcohol at or near the boiling point of same, as shown below:

X .0 )7 /^r2 // l ⁺ ®k ethanol \_/ ^XO^{? X}R₃ δ z

⁵ L _(I) . // /^CH-CH^ .

L— ^0H wherein X and Z are halogen, R₂ and R₃ are as hereinabove defined and Y is hydrogen. Thus, 3,5-dichlorostyrene oxide can be reacted with an equimolar or molar excess of t-butylamine in ethanol at reflux from about 1 to about 8 hours, or until the reaction is essentially complete and the de25 sired a-[(t-butylamino)methyl]-3,5-dichlorobenzyl alcohol is obtained as illustrated below:

Cl Cl

<img file="IL60530A_D0008.tif" />

Cl Cl

The thus obtained product can be purified by known procedures, such as chromatography or recrystallization of salt ־.hereof.

/2 ר then isolated by־׳sta’nd^^^. if so desired. The above ' illustrated as follows:

¾. The thus obtained halo compound is isolated'; by cbr^r ventional laboratory methods and is then reacted with the&lt; . appropriate alcohol, under an inert blanket of gas, siich^s^X^ nitrogen at a temperature range of from about 0 to 5°C.

thus obtained formula (I) product is ard laboratory methods and purified, reaction sequence may be graphically

X

<img file="IL60530A_D0009.tif" />

ζ

CH-CH-NR-R-

3 2 2 ן

OH

SOC1₂

X

<img file="IL60530A_D0010.tif" />

Ζ

CH-CH₂NR₂R₃

R_COH

ס

X

<img file="IL60530A_D0011.tif" />

Ζ ch־ch₂nr₂r₃

OR_C wherein X,Y.Z,R₂,R₃ and Rg are as hereinabove defined.

Alternatively, a formula (I) compound wherein

R^ is ORg may be prepared by dissolving the corresponding formula (I) compound wherein R^ is OH in the corresponding R_COH alcohol and saturating the thus obtained solution with dry HC1 gas. The reaction mixture is then stirred at room temperature for a period of time sufficient to essentially complete the reaction and the product is then isolated by standard laboratory procedures and purified, if so desired. This reaction sequence may be illustrated as 35 follows:

S0/2

<img file="IL60530A_D0012.tif" />

wherein X,Y,Z,R₂,R₃ and Rg are as hereinabove defined.

In the present specification and claims the term a,a-dimethy!phenethyl means a structure having the following configuration:

<img file="IL60530A_D0013.tif" />

When orally administered in the feed, generally about 0.01 to 300 grams per ton of feed of the aboveidentified phenylethanolamine derivative or acid addition 35 salt thereof, is effective for enhancing the growth rate and improving the efficiency of feed utilization by the above-mentioned meat-producing animals.

60530/3 ־ 6.-Since the effective and preferred dietary levels of the active ingredient vary somewhat from species to species in the above-mentioned animals said level's for each animal species are listed in Table I below on a gram per ton of feed basis:

Table I

<td> Compound</td><td> Effective Feed Level g/ton</td><td> Preferred Level g/ton</td><td> Animal</td>

<td rowspan="5"> Formula (I)</td><td> 0.1 -300</td><td> 200־1</td><td> Swine</td>

<td> 0.1 -200</td><td> 1-100</td><td> Sheep, Goats</td>

<td> 0.01-50</td><td> 0.1-10</td><td> Chickens, Rabbits</td>

<td> 50־0.01</td><td> 0.1-10</td><td> Turkeys</td>

<td> 200־ 0.1‘</td><td> 1-100</td><td> Cattle</td>

Animal feed compositions which will provide the desired growth promotion and feed efficiency in the above-mentioned animals can be prepared by admixing the phenylethanolamine derivative or acid addition salt thereof, or an animal feed supplement containing said compound, with a sufficient ״quantity of an appropriate animal feed to provide the desired level of active • -J compound in said feed.

Animal feed supplements can be prepared by admixing about 75% to 95% by weight of the phenylethanolamine derivative or acid addition salt thereof, with about 5% to 25 25% by weight of a suitable carrier or diluent. Carriers suitable for use to make up the feed supplement compositions include the following: alfalfa meal, soybean meal, cottonseed oil meal, linseed oil meal, sodium chloride, cornmeal, cane molasses, urea, bone meal, corncob meal and the like. ₃q The carrier promotes a uniform distribution of the active ingredient in the finished feed into which the supplement is blended. It thus performs an important function by ensuring proper distrubition of the active ingredient throughout the feed.

If the supplement is used as a top dressing for feed, it likewise helps to ensure uniformity of distribution of the active material across the top of the dressed feed.

For parenteral administration the phenylethanol' jl

־ 7 ־ amine derivative may be prepared in the form of a paste or pellet and administered as an implant, usually under the skin of the heat or ear of the animal in which enhanced growth rate and/or improved efficiency of feed utilization is 5־ sought.

In practive, parenteral administration generally involved injection of a sufficient amount of the above-said ethane derivative to provide the animal with from 0.001 to 100 mg/kg of body weight of the active ingredient. The pre10 ferred dosage level for swine is 0.01 to 50 mgAg of body weight and for cattle the range of from 0.001 to 50 mg/kg of body weight of the active phenylethanolamine derivative is preferred. The preferred dose level of said ethane derivative for poultry is about 0.001 to 35 mg/kg of animal 15 body weight and the preferred dose level of said ethanol derivative for sheep and goats is 0.001 to 40 mg/kg of animal body weight. The preferred dose level for rabbits and domestic pets is 0.001 to 35 mg/kg of animal body weight.

Paste formulations can be prepared by dispersing 20 the active ethanol derivative in a pharmaceutically acceptable oil such as peanut oil, sesame oil, corn oil or the like.

Pellets containing an effective level of the phenylethanolaraine derivative can be prepared by admixing the above-said active ingredient with a diluent such as carbowax, 25 biodegradable polymers, carnuba wax, or the like. A lubricant, such as magnesium stearate or calcium stearate may be added to improve the pelleting process if desired.

It is, of course, recognized that more than one pellet may be administered to an animal to achieve the de30 sired dose level which will provide the increased growth rate and/or improved efficiency of feed utilization by said animal. Moreover, it has been found that additional implants may also be introduced periodically during the treatment period in order to maintain the proper drug release rate in 35 the animal's body.

In addition to enhanced growth promotion and improved efficiency of feed utilization by meat-producing animals, the compounds of the present invention have the add ed advantage that, at selected levels of administration they depress the deposition of fat in said animals. This biological response has substantial advantage to poultrymen and swine producers since administration of said compounds at selected levels yields leaner animals which command premium prices from the meat industry.

The invention has several advantages for the pet owner or veterinarian who wishes to trim unwanted fat from pet animals. For poultry men and swine raisers, using the method of the present invention were attained increased yields of leaner animals which command higher prices from the meat industry. Surprisingly, it is also noted that feed efficiency and animal growth rate are significantly enhanced when the compounds of the present invention are administered to swine and poultry at selected dose levels.

The following examples illustrate the invention.

Example

Evaluation of test compounds as animal growth promoters

CF1 female mice from Carworth Farms are received when they are six weeks old. They are housed ten to a cage in air-conditioned rooms (72^eF to 76°F) with automatically controlled lights, 14 hours on and 10 hours off. The basal diet used in these studies is Purina Laboratory Chow (see description below), which is supplied ad libitum. Water is also allowed ad libitum.

Thirteen days after arrival, the mice are weighed in groups of ten and assigned at random to the different treatments. The concentration of the different compounds in the diet is indicated in the following tables. Twelve days later the mice are weighed again and the experiment terminated. Test data are provided in Table II below wherein data are reported as percentage gain over controls. The following is a description of the diet to which the growth-promoting compounds were added.

DIET

Guaranteed Analysis

Crude protein not less than 23.0%

Crude fat not less than 4.5% χ ....

- W530/1

. ' ,'4 \ _־\

Crude fiber not more than 6.0%\ X

Ash not more than 9.0% \ ' ζ

IngredientsXxf

׳. \

Meat and bone meal, dried skimmed milk, wheat germ meal, - Vx 5 fish meal, animal liver meal, dried beet pulp, ground extraded corn, ground oat groates, soybean meal, dehydrated alfalfa meal, cane molasses, animal fat preserved with B^A, vitamin supplement, calcium pantothenate, choline chloride, folic acid, riboflavin supplement, brewer's dried yeast, thiamin, niacin, vitamin A supplement, D-activated plant sterol, vitamin E supplement, calcium carbonate, dicalcium phosphate, iodized salt, ferrec ammonium citrate, iron oxide, manganous oxide, cobalt carbonate, copper oxide, zinc oxide.

TABLE

Evaluation of Teat Coapounda as Aniaal Growth Promoters

Initial Final Gain

House Mouse Gain Over

SLiil , Wt.,(g) (grama) Control

<td colspan="4"> ^-Amino-g-(tert-butylamin omethy1)-3,5-dichlorobenzy1 alcohol hydrochloride</td>

<td> (ppm in diet)</td><td></td><td></td><td></td>

<td> 0</td><td> 23.116</td><td> 29.67</td><td> 1.21</td>

<td></td><td> 23.96</td><td> 29.99</td><td> 1.03</td>

<td></td><td> 22.79</td><td> 29.62</td><td> 1.83</td>

<td></td><td> 29.10</td><td> 25.98</td><td> 1.88</td>

<td></td><td> 29.23</td><td> 25.52</td><td> 1.29</td>

<td></td><td> 23.63</td><td> 29.93</td><td> 1.30</td>

<td></td><td> 23.33</td><td> 29.76</td><td> 1.93</td>

<td></td><td> 22.75</td><td> 23.86</td><td> 1.11</td>

<td> Control Average</td><td> 23.97</td><td> 29.85</td><td> 1.39</td>

TABLE (Continued)

Evaluation of Test Compounds aa Animal Growth Promoters

<td></td><td></td><td></td><td></td><td> ί</td>

<td></td><td> Initial</td><td> Final</td><td></td><td> Gain</td>

<td></td><td> House</td><td> House</td><td> Gain</td><td> Over</td>

<td> Dosage</td><td> Ol'</td><td></td><td> (grama)</td><td> Control</td>

<td> 50</td><td> 22.95</td><td> 25.63</td><td> 2.68</td><td></td>

<td></td><td> 23.91</td><td> 26.19</td><td> 2.23</td><td></td>

<td></td><td> 21.26</td><td> 26.30</td><td> 2.09</td><td></td>

<td> Average</td><td> 23.71</td><td> 26.02</td><td> 2.32</td><td> +66.9</td>

<td> 100</td><td> 23.50</td><td> 25.39</td><td> 1.89</td><td></td>

<td></td><td> 23.80</td><td> 26.04</td><td> 2.29</td><td></td>

<td></td><td> 23.00</td><td> 25.65</td><td> 2.65</td><td></td>

<td> Average</td><td> 23.93</td><td> 25.69</td><td> 2.26</td><td> +62.6</td>

<td> 200</td><td> 23.03</td><td> 24.80</td><td> 1.77</td><td></td>

<td></td><td> 29.50</td><td> 26.12</td><td> 1.62</td><td></td>

<td></td><td> 23.08</td><td> 25.09</td><td> 1.96</td><td></td>

<td> Average</td><td> 23.59</td><td> 25.32</td><td> 1.78</td><td> +28.1</td>

60530/2

The procedure described above is repeated using control animals for each test. Twelve days after the tests are started the animals are weighed and the test terminated. The results of each test are reported in Table III below as weight gains for each test group and percent gain for each group over controls.

TABLE

Evaluation of Teat Compounds as Animal Growth Promoters

<td></td><td> Dosage</td><td> Gain</td><td> } Gain Over</td>

<td> Compound</td><td> (wi</td><td> (grans)</td><td> Controls</td>

<td> 4-taino-3,5־dlbr0(i01&gt;־-[(tert-butylaaln0)!1ethyll-</td><td> 400</td><td> 16.5</td><td> +22.2</td>

<td> benzyl alcohol hydrochloride</td><td> 200</td><td> '20.4</td><td> +51.1</td>

<td></td><td> 100</td><td> 22.9</td><td> +69.0</td>

<td></td><td> 50</td><td> 23.3</td><td> +72.6</td>

<td> 1! - Am ino-3₁5-dibromo-a-[ (di Isopropy lam Ino) me thy 1 ] .</td><td> 200</td><td> 20.2</td><td> +46.4</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 16.9</td><td> +22.5</td>

<td></td><td></td><td></td><td> i</td>

<td> P-amino-a-[(dimethylamino)methyl]benzyl alcohol</td><td> 200</td><td> 17.6</td><td> +28.3</td>

<td></td><td> 100</td><td> 15.6</td><td> +13.0</td>

<td> P-amino-a-[(diisopropylaminojmethyl|benzyl alcohol</td><td> 200</td><td> 16.5</td><td> +18.7</td>

<td> hydrochloride</td><td> 100</td><td> 14.2</td><td> + 2.2</td>

<td> i’Amino-3,5-dichloro־a-[(dimethylamino)methyl|benzyl</td><td> 100</td><td> 18.4</td><td> + 7.6</td>

<td> alcohol hydrochloride</td><td></td><td></td><td></td>

<td>¹l-Amino-J^-dichloro-a-lidiisopropylaminoimethyl]־׳</td><td> 200</td><td> 23.9</td><td> +66.0</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 21.3</td><td> +47.9</td>

BZQSS09 )

TABLE (Continued)

Evaluation of Teat Compounds an Animal Growth Promoters

<td></td><td> Dosage</td><td> Gain</td><td> I Gain Over</td>

<td> Compound</td><td> (ppm)</td><td> (grams)</td><td> Controls</td>

<td>¹l-Aniino-JiS-dichloro-arKcyclohexylaminolmethyl]-</td><td> 200</td><td> 19.3</td><td> +48.5</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 16.3</td><td> +25.4</td>

<td> P-amino-a-[(tert-butylamino)methyl)benzyl alcohol</td><td> 200</td><td> 19.6</td><td> +88.5</td>

<td></td><td> 100</td><td> 18.2</td><td> ♦75.0</td>

<td></td><td> 50</td><td> 17.9</td><td> +72.1</td>

<td></td><td> 25</td><td> 13.6</td><td> +30.8</td>

<td> 4-Amino-3,5-dichloro-a-((methylamlno)methyl]benzyi</td><td> 200</td><td> 15.6</td><td> +54.5</td>

<td> alcohol hydrochloride</td><td> 100</td><td> 18.9</td><td> ♦87.1</td>

<td> P-amino-a.:[ (methylamino)methyl|benzyl alcohol hydrochloride</td><td> 100</td><td> 16.8</td><td> +18.4</td>

<td> a-(+talno-3,5-dichloropheny!H^orphollneethanol</td><td> 200</td><td> 15.9</td><td> *34.7</td>

<td> hydrochloride</td><td> 100</td><td> 14.0</td><td> +18.6</td>

<td> !|-A1nino-a-[(sec-butylamino)methyll-315-dichlorobenzyl alcohol</td><td> 200</td><td> 13.2</td><td> +16.8</td>

C/OSS09

TABLE HI (Continued)

Evaluation of Teat Compounds as Animal Growth Promoters

<td></td><td> Dosage</td><td> Gain</td><td> ί Gain Over</td>

<td> Compound &gt; a</td><td> (ppm)</td><td> (grama)</td><td> Controls</td>

<td> 4-talno3,5־-dlchloro-e- [(-3-methoxypropyl)amlno)-</td><td> 200</td><td> 15.4</td><td> +23.2</td>

<td> methyl benzyl alcohol</td><td> 100</td><td> 23.0</td><td> +84.0</td>

<td> 4-Aaino-3,5-dlchloro-a-((diallylamino)methyllbenzyl</td><td> 200</td><td> 17.4</td><td> +39.2</td>

<td> alcohol hydrochloride</td><td> 100</td><td> 18.4</td><td> +47.2</td>

<td> i-Amlno-3״5_׳dlchloro-a-[(benzylamino)methyl] benzyl alcohol hydrochloride</td><td> 200</td><td> 10.6</td><td> +19.1</td>

<td> 4-talno-a-[(butylamlnolmethyl]-3,5-dlchlorobenzyl alcohol</td><td> 100 .</td><td> 14.6</td><td> +64.0</td>

<td>¹!-Amino-SiS-dlchloro-a-lfll-iDethyl-l-plperazinyl)-</td><td> 200</td><td> 9.4</td><td> + 5.6</td>

<td> methyl)benzyl alcohol</td><td> 100</td><td> 9.5</td><td> + 6.7</td>

<td> i-Anjlno-3,5-dlchloro-a״[(lsopropylamino)methyl|-</td><td> 200</td><td> 13.1</td><td> +26.0</td>

<td> benzyl alcohol</td><td> 100</td><td> 18.9</td><td> +81.7</td>

TABLE (Continued)

Evaluation of Test Compounds as Animal Growth Promoters

<td></td><td> Dosage</td><td> Gain</td><td> f Cain Over</td>

<td> Compound</td><td> (ppm)</td><td> (grams)</td><td> Controls</td>

<td> i|-Amino-a־(ai11nofflethyl)-3j5־dichlorobenzyl alcohol hydrochloride</td><td> 100</td><td> 13.4</td><td> +19.0</td>

<td>¹J-Araino-3 _t5-dichloro-a-[ (hexylaminojmethy 1| benzyl</td><td> 200</td><td> 16.8</td><td> +15.9</td>

<td> alcohol</td><td> 100</td><td> 19.2</td><td> +32.8</td>

<td> a.־( (tert״butylamino)methylN3.5״dichlorobenzyl</td><td> 200</td><td> 19.3</td><td> +33.1</td>

<td> alcohol hydrochloride</td><td> 100</td><td> 20.2</td><td> +39.3</td>

<td> 4-Amino-3,5-dichloro-a-[(dlethylamino)methyl]benzyl alcohol hydrochloride</td><td> 100</td><td> 17.5</td><td> , +20.1</td>

<td> (allylamino)methyl]-4-an11no-315“dichlorobenzyl</td><td> 200</td><td> 16.8</td><td> +118.2</td>

<td> alcohol</td><td> 100</td><td> 17.!</td><td> +122.1</td>

<td> 4-Amino-a־(anilino1nethyl)-3,5-dichlorobenzyl alcohol</td><td> 200</td><td> 20.7</td><td> +25.5</td>

<td></td><td> 100</td><td> 17.5</td><td> 16.1</td>

TABLE (Continued)

Evaluation of Teat Compounda aa Animal Growth Promoters

<td></td><td> Dosage</td><td> Gain</td><td> ( Gain Over</td>

<td> Compound</td><td>¹</td><td> (grams)</td><td> Controls</td>

<td> l-faino-a-[1-tert-butylamino)ethyl]-3!5-dichloro-</td><td> 200</td><td> •22.7</td><td> +37.6</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 23.8</td><td> +99.5</td>

<td> 9-talno-3-bromo-«-[(tert-butylamino)11ethyl|-5־chloro-</td><td> 200</td><td> 18.5</td><td> +60.9</td>

<td> benzyl alcohol hydrochloride</td><td> 100</td><td> 19.5</td><td> ♦69.6</td>

<td> «1־(tert-butylamino)methy11 nn-hydroxybenzyl alcohol</td><td> 200</td><td> 15.8</td><td> ♦ 9.0</td>

<td> hydrochloride</td><td> 100</td><td> 19.9</td><td> +37.2</td>

<td> a-[(iaopropylaaino)11ethyl]-e1־hydroxybenzyl alcohol</td><td> ¢00</td><td></td><td> ♦59.9</td>

<td> hydrochloride</td><td> 200</td><td></td><td> +53.0</td>

<td></td><td> 100</td><td> *J</td><td> ♦39.7</td>

<td> a-[(Amlno)methyl]-m-hydroxybenzyl alcohol hydrochloride</td><td> 200</td><td></td><td> + 9.9</td>

<td></td><td> 100</td><td> 1</td><td> +30.7</td>

<td> 4 - Amin0-N־tert-butyl3,5־‘dichl0r0“Heth0xyphenethyl</td><td> 200</td><td> 18.8</td><td> +37.2</td>

<td> hydrochloride</td><td> 50</td><td> 21.6</td><td> +57.7</td>

Z/O2SO9 ¢0530/2 ־ 18 Example

Evaluation of test compounds as animal growth promoters

The procedure of Example 1 is used in this evaluation. The diet is the same as described in said example and data obtained are reported as percent gain over controls. Data are reported in Table IV below.

)

TABLE

Evaluation of Test Ccmpounda as Animal Growth Promoters

<td> Dosage</td><td> Initial !LISI</td><td> Final !Liil</td><td> Gain (grama)</td><td> ί Gain Over Control</td>

<td> 4-Amlno-a-(tert-butylaminonethyl)- 3,5-dichlorobenzyl alcohol hydrochloride</td><td></td><td></td><td></td><td></td>

<td> (ppm In diet)</td><td></td><td></td><td></td><td></td>

<td> 0</td><td> 24.49</td><td> 25.17</td><td> .68</td><td></td>

<td></td><td> 21.25</td><td> 26.06</td><td> 1.81</td><td></td>

<td> 1</td><td> 23.65</td><td> 25.13</td><td> 1.78</td><td></td>

<td></td><td> 22.83</td><td> 24.33</td><td> 1.50</td><td></td>

<td></td><td> 24.39</td><td> 25.59</td><td> 1.20</td><td></td>

<td></td><td> 24.36</td><td> 26.06</td><td> 1.70</td><td></td>

<td></td><td> 23.11</td><td> 24.50</td><td> 1.39</td><td></td>

<td></td><td> 23.54</td><td> 24.82</td><td> 1.28</td><td></td>

<td> Average</td><td> 23.83</td><td> 25.25</td><td> 1.12</td><td></td>

Z/OCS&amp;9 )

TABUS (Continued)

Evaluation of Test Compounds as Animal Growth Promoters ί

<td></td><td> Dosage</td><td> Initial wt. ω</td><td> Final Mt. (g)</td><td> Gain (grams)</td><td> Gain Over Control</td>

<td> 200</td><td></td><td> 23.46</td><td> 25.49</td><td> 2.03</td><td></td>

<td></td><td></td><td> 23.68</td><td> 25.97</td><td> 2.37</td><td></td>

<td></td><td></td><td> 23.56</td><td> 25.40</td><td> 1.84</td><td></td>

<td> Average</td><td></td><td> 23.57</td><td> 25.62</td><td> 2.05</td><td> +44.4</td>

<img file="IL60530A_D0014.tif" />

־ 21 ־

Example

Evaluation of test compounds as animal feed additives for the enhancement of the growth rate of poultry

One day old Hubbard X Hubbard Crossbred Chicks, randomly allotted to pens of ten chicks (5 males and 5 females) each.

Eight pens of chicks are used for unmedicated controls, and four pens of chicks are used at each level of drug. The duration of the experiment is 28 days.

The controls are offered an unmedicated diet of

Broiler Ration No. 453 (composition given below) and water ad libitum. Medicated chicks are offered the same diet containing the test drug at the levels indicated above, and water ad libitum. The weight of the chicks is determined at the beginning and on completion of the experiments. Weight gains and the amount of feed consumed are also determined. The thus obtained data are averaged and summarized in Table V below, wherein the percent improvement in weight gains and feed/gain ratios are given.

<td> 20</td><td> Component</td><td> Percent by Weight</td>

<td> 25</td><td> Ground yellow com Soybean oil meal (49%) Menhaden fish meal (60%) Corn gluten meal (60%) Dehydrated alfalfa meal (17%)</td><td> 53.45 28.00 5.0 5.00 2.00</td>

<td> 30</td><td> Stabilized fat Dicalcium phosphate Ground limestone Sodium chloride Trace minerals mixture*</td><td> 4.00 1.20 0.50 0.30 0.05</td>

<td> 35</td><td> Vitamin premix.* *Trace Mineral Mixture Manganese 12.50 % Iron 6.00 Zinc 5.00 Copper 0.65</td><td> 0.50 100.00 1 lb/ton furnishes 62.5 ppm 30.0 25.0 3.25</td>

Iodine 0.351.75

Cobalt 0.251.25

Calcium minimum15.30

Calcium maximum17.35

<td> 5</td><td> *♦Vitamin Premix for 1-ton</td><td> Weight in Gram</td>

<td></td><td> DL Methionine</td><td> 453.6</td>

<td></td><td> BHT (butylated hydroxy toluene)</td><td> 113.6</td>

<td></td><td> Vitamin A (30,000 mcg/g)</td><td> 100.0</td>

<td></td><td> Vitamin (200,000 mcg/g)</td><td> 5.0</td>

<td> 10</td><td> Vitamin E (20,000 mcg/lb)</td><td> 45.4</td>

<td></td><td> Riboflavin</td><td> 4.0</td>

<td></td><td> Niacinamide</td><td> 25.0</td>

)

TABLE

Mean Weight Gain and Feed Efficiency of Control and Test Compound - Treated Chicks

Thirteen-Day Battery Testing ppm Mean t J Improvement

<td> Treatment</td><td> in Diet</td><td> Gain (g)</td><td> Control</td><td> F/G</td><td> over Control</td>

<td> Control</td><td> 0</td><td> 266.5</td><td> «Β</td><td> 1.90</td><td></td>

<td> 4-Amino-a-[(tert-butylamino)methyll</td><td> 0.3</td><td> 271.8</td><td> +2.0</td><td> 1.90</td><td> 0</td>

<td> 3,5-dlchlorobenzyl alcohol</td><td> 0.6</td><td> 274.3</td><td> ♦2.9</td><td> 1.38</td><td> +1.¢</td>

<td></td><td> 1.25</td><td> 259.9</td><td> 2.5</td><td> 1.91</td><td> ?.ס-</td>

<td></td><td> 2.5</td><td> 260.6</td><td> 2.1</td><td> 1.39</td><td> +0,?</td>

<td></td><td> 5.0</td><td> 251.9</td><td>.5</td><td> 1.90</td><td> 0</td>

<td> ^Amino-]₍5-dibro01o-a-[ (tert-butyl-</td><td> 0.3</td><td> 270.8</td><td> +1.6</td><td> 1.90</td><td> 0</td>

<td> aminojmethyljbenzyl alcohol hydrochloride</td><td> 0.6</td><td> 271.8</td><td> +2.0</td><td> 1.38</td><td> +1.¢</td>

<td></td><td> 1.25</td><td> 267.5</td><td> +0.11</td><td> 1.39</td><td> +0.?</td>

<td></td><td> 2.5</td><td> 265.2</td><td>.5</td><td> 1.38</td><td> +1.¢</td>

<td></td><td> 5.0</td><td> 270.3</td><td> +1.¢</td><td> 1.37</td><td> +2.1</td>

Z/OSS99

40S39/2

Example 4

Evaluation of test compounds as antilipogenic agents Mouse tests

CFI female mice from Carworth Farms are received when they are six weeks old. They are housed ten to a cage in air-conditioned rooms (72°F to 76°F) with automatically controlled lights, 14 hours on and 10 hours off. The basal diet used in these studies is Purina Laboratory Chow (see description below), which is supplied ad libitum.

The following is a description of the diet to which the growth-promoting compounds were added.

DIET

Guaranteed

<td></td><td> Crude protein not less than</td><td> 23.0%</td>

<td> 15</td><td> Crude fat not less than</td><td> 4.5%</td>

<td></td><td> Crude fiber not more than</td><td> 6.0%</td>

<td></td><td> Ash' not more than</td><td> 9.0%</td>

Ingredients

Meat and bone meal, dried skimmed milk, wheat germ meal, 20 fish meal, animal liver meal, dried beet pulp, ground extruded corn, ground oat groats, soybean meal, dehydrated alfalfa meal, cane molasses, animal fat preserved with: BHA, vitamin B₁₂ supplement, calcium pantothenate, choline chloride, folic acid, riboflavin supplement, brewer's dried 25 yeast, thiamin, niacin, vitamin A supplement, D-activated plant sterol, vitamin E supplement, calcium carbonate, dicalcium phosphate, iodized salt, ferric ammonium citrate, iron oxide, manganous oxide, cobalt carbonate, copper oxide, zinc oxide. Water is also allowed ad libitum.

Thirteen days after arrival, the mice are weighed in groups of ten and assigned at random to the different treatments. The concentration of the different compounds in the diet is indicated in the following tables. Twelve days later the mice are weighed again the experiment ter35 minated. At least three cages (30 mice) of untreated controls are included in each test. Test data are provided in Table VI below wherein data are reported as percent body fat, percent change in body fat from controls and gain per mouse

TABLE V

AntUipogenio Agent Evaluation and Growth Enhancement Evaluation in Mice

Level Number Average Average Gain Change in of Mice Initial Final per { in 1

Diet per Weight Height House Body Fat from Compound (ppm) Treatment (g) (g) . (g) Fat Control

11.95 1.9 25.0 23.6 50 0 1ן

H ^.XcH-NH־C(CH) ^{50 3}° ²⁶’° ^{2,3 55}'^״ ° .=. 100 3o 23.9 25.1 2.3 10.23 •19.90 ¹ 200 30 23.5 25.3 1.8 10.50 12.13

900 30 23.9 29.9 1.9 9.10 -23.80

Percent Body Fat Determination of Mice

A. Preparation of Carcasses:

Stomach and intestines are removed from each mouse. All other viscera, including skin and fur, remain intact. Each cage of mice (10) are weighed and added to a 1000 ml beaker and autoclaved at 120°C (15 psi) for 30 minutes. Carcasses from each cage are then blended and homogenized. The homogenate is weighed and duplicate 5-gram samples are removed for analysis.

B. Fat Analysis:

Fifteen milliliters (ml) of concentrated hydrochloric acid is added to each 5-gram sample and mixed well. Samples are heated in an 84 ^eC water bath for 2 hours. To extract the fat, thirty ml of petroleum ether is added to each samples, 15 ml at a time, and mixed well on a Vortex mixer. The aqueous and organic phases are separated by low speed centrifugation and the ether layer (containing fat) is extracted into tared 30 ml beakers. After evaporating to dryness the beaker containing fat is reweighed to determine grams of fat per five grams of homogenate. Total body fat in the carcass is calculated as follows:

<td> % Fat = —</td><td> grams fat in sample</td><td></td><td> grams total homogenate</td><td></td>

<td></td><td> gram weight' of sample</td><td></td><td colspan="2"> --------—— ׳ X 1UU carcass weight “ of mice (g)</td>

Example Antilipogenic Evaluation of test compounds - Mouse Study

CFI female mice, 55 days old, are weighed in groups of 10 and allotted to cages to minimize weight variation among cages. Treatments are randomly assigned to cages.

Each of the treatments are tested in 3 replicates,

i.e., in 3 cages of 10 mice each. There are 10 cages of 10 control mice each. Durgs are mixed in the diet at the dosage level indicated. Feed and water are offered ad libitum for the 12-day test period. Feed spilled is collected during the test period. At the end of the test period, the collected feed is weighed and the mean feed consumption per cage of ten mice is determined for each treatment. The mice are weighed as a group of 10 and the weight gain determined. The mice are sacrificed by cervical dislocation. The right uterine fat pad of each mouse is removed. The fat pads for each cage of 10 mice are weighed as a unit.

To establish correlation between the percent reduction in fat pad weights of treated animals and percent ’ reduction in total body fat of treated animals, animals from several treatment groups are evaluated for total body fat using the body fat determination described in Example 5. Data obtained are reported in Table VII for those groups upon which such determination has been made. From percent reduction in fat pad weight and the total fat determinetions for the groups tested, it can be seen that a reduction in fat pad weights of animals is generally indicative of a reduction of total body fat of the treated animals.

TABLE Ώ,

Antilipogenic Evaluation of Test Compounds - Mouse Study

J Reduction in Fat

<td> Compound</td><td> Doeage (ppm) »</td><td> Pad Height vs Controls</td><td> J Animal Fat vs Controls</td>

<td> 1-amlno-3,5-dlbromo-a-[(tert-butylamino)methyl]benzyl</td><td> ¢00</td><td>.1</td><td>.6</td>

<td> alcohol hydrochloride</td><td> 200</td><td> ’ 27.5.</td><td>.1.</td>

<td></td><td> 100</td><td> •13.9*</td><td>.3*</td>

<td> 4-amin0-3,5־dlbr0in0-a-|(diisopropylamino)methyl|benzyl alcohol hydrochloride</td><td> 200</td><td>.1</td><td>.1</td>

<td> 9-amino-a-Ktert-butvlamlno)methvll-3.5-dlchlorobenzvl</td><td> ¢00</td><td>.0</td><td>.8</td>

<td> alcohol hydrochloride</td><td> 200</td><td>.1</td><td>.1</td>

<td></td><td> 100</td><td>.9</td><td> 11.1</td>

<td> *l-a1nlno-3»5-dichloro-a-[(methylan1ino)n1ethylJbenzyl</td><td> 200</td><td>.7</td><td>.9</td>

<td> alcohol hydrochloride</td><td> 100</td><td>.8</td><td>.5</td>

<td> *t-amino-ifb-dichloro-a[(diethyl)methyl)benzyl alcohol</td><td> 200</td><td>.7</td><td></td>

<td> hydrochloride</td><td> 100</td><td>.2</td><td></td>

<td> *l-amino-a-[(sec-butyla1nino)1nethyl]-3,5-dichlorobenzyl</td><td> 200</td><td>.2</td><td>.3</td>

<td> alcohol</td><td> 100</td><td>.2</td><td>.3</td>

*Average 2 tests

TABLEVIlf (Continued) dntilipogenic Evaluation of Teat Caipounda - House Study

<td> 1 Compound</td><td> Dosage (»).</td><td> J Reduction in Fat Pad Weight vs Controls</td><td> J Animal Fat va Controls</td>

<td> lamino-3,5-dichloro-a- ((diallylamlno)methyl| benzyl alcohol hydrochloride</td><td> 200</td><td> . -12.0</td><td></td>

<td> 4-amino-3p5״dichloro-a-[(benzylamino)methyljbenzyl</td><td> 200</td><td>.7</td><td>.9</td>

<td> alcohol hydrochloride</td><td> 100</td><td>.1</td><td> 1.7</td>

<td> t-amino-a-[(butylamino)methyl|-3»5-dichlorobenzyl</td><td> 200</td><td>.59</td><td>.78</td>

<td> alcohol</td><td> 100</td><td>.7</td><td> 13.08</td>

<td> 4-amino-3,5-dichloro-a-[(i3opropylamino)n!ethylJbenzyl</td><td> 200</td><td>.2</td><td>.5</td>

<td> alcohol</td><td> 100</td><td>.9</td><td> 20.9</td>

<td> a-[(allyla111ino)methyl|-i|-amino-3|5-dichlorobenzyl</td><td> 200</td><td>.5</td><td></td>

<td> alcohol</td><td> 100</td><td>.3</td><td></td>

<td> Vamlno-a-[1-(tert-butylamino)ethyll3,5־-d ichlorobenzvl alcohol hydrochloride</td><td> 100</td><td>.8</td><td></td>

<td> a-[(t־butylamino)methyl]-3,5-dichlorobenzyl</td><td> 200</td><td>.5</td><td></td>

<td> alcohol hydrochloride</td><td> 100</td><td>.8</td><td></td>

Antilipogenlc Evaluation of Test Compounds -Mouse Study

<td></td><td> Z Reduction in Fat</td>

<td> Dosage Compound (ppm)</td><td> Pad Weight Z Animal Fat vs Controls vs Controls</td>

<td> 4-amino-3-bromo-a-[(t-butylamino)methylj-5-chloro- 200 benzyl alcohol hydrochloride 100</td><td>.8 -18.7</td>

<td> m-hydroxy-a-Kisopropylamino)methyl]benzyl alcohol 400 200 100</td><td>,8 -26,2 -7.5</td>

<td> 4-amino-N-t-butyl-3,5-dichlor ophene thy lamine 50 hydrochloride</td><td> • •24.8</td>

<td> 4-amino-3,5-dichlor0-a-[ (cyclopropylamino)methylj- 100 benzyl alcohol</td><td>,7</td>

<td> 4-[2-(t-butylamino)-l-hydroxyethyl]-2’-chloroacetanilide 200 100</td><td> 6.7- 12.1־</td>

<td> 4-amino-3,5-dichlor0-a-[ (cyclopentylamino)methyl]benzyl 200 alcohol 50</td><td>.5 -4.7</td>

<td> 4-amino-3,5-dichloro-cHI (2-hydroxyethyl) aminoJmethyl)- 200 benzyl alcohol 50</td><td>,2 -7.4</td>

<td> 4-amino־a-[(t-butylamino)methyl]-3,5-diiodobenzyl alcohol 200 hydrochloride 100</td><td>.6 -26.6</td>

<td> 4-amlno-H-butyl-3,5-dichloro-J-methoxyphenethylamine 200 hydrochloride 50</td><td>.4 -21.7</td>

Example 6

Antilipogenic evaluation of test compounds - Rat study The procedure employed and the diet used for evaluation of test compounds as antilopogenic agents mice, are described in Example 1, excepting that the treatment period is fourteen days and 10 rats, one per cage, are used for each treatment.

Percent body fat is determined in the same manner as described in Example 4, excepting that the skin and organs 10 are removed before the carcasses are homogenized.

Results of this study are reported in Table VIII below.

TABLE VII ,

Antilipogenic Evaluation of Teat Compounda and Growth Rate Evaluation in Rats

<td colspan="2"> Level</td><td> Number</td><td> Average</td><td> Average</td><td> Average</td><td colspan="2"> Change</td>

<td></td><td> in</td><td> of Rats</td><td> Initial</td><td> Final</td><td> Gain</td><td> J Fat in</td><td> in Fat</td>

<td></td><td> Diet</td><td> per</td><td> Weight</td><td> Weight</td><td> per Rat</td><td> Eviscerated</td><td> from</td>

<td> Compound</td><td> (ppm)</td><td> Treatment</td><td> (g)</td><td> JiL</td><td> (g)</td><td> Carcass</td><td> Control</td>

4.67 76.4 149.1 ’ 72.7 10 0ף /Λ , ₍ 25 10 ?8.3 159.3 81.0 3.09-39.9

I K Mh-CH -NH-i(CH) .HC1 w 100 9 ?6.6 159. 8 83.2 2.5992.0

95.6 2.71 3.2? 196.9 3.2? 10 900׳י®

S/OS-SO9 ¢0510/2

EXAMPLE 7

Evaluation of teat compounds as animal feed additives for the enhancement of growth rate and improvement in feed efficiency of mice.

Four-week old female outbred rats (5-gram range) from Charles River Breeding Laboratories, 251 Ballardvale Street, Wilmington, Massachusetts 01887, are housed 2/cage in air-conditioned rooms (72°F to 76°F) with automatically controlled lights, 14 hours on and 10 hours off. The basal 10 diet used in these studies is Purina Laboratory Chow which is supplied ad libitum. Water is also given ad libitum.

Four days after arrival, the animals are weighed and allotted to treatment groups to minimize weight variation. Ten rats are used per treatment group. Drugs are 15 administered in the feed at 2 ppm, 10 ppm and 50 ppm for a period of 12.5 weeks. Animals are weighed weekly and feed consumption corrected for spillage recorded daily. The results of this trial are shown below in Table IX.

TABLE _

Evaluation of Teat Compounds as Animal Feed Additives for the Enhancement of Growth Bate and Improvement In Feed Efficiency - Mice

Feed _t

<td> Treatment</td><td> Dose ppm</td><td> Gain^aliL</td><td> Consumption^ ...(81.,.</td><td> Feed/Gain J Improvement</td>

<td> Control</td><td></td><td> 157</td><td> 1301</td><td> 8,31</td>

<td> i|-Amino-a-(tert-butyla111ino1Dethyl)-3,5־dichloro-</td><td> 2</td><td> 178 (+13.M)</td><td> 1M3 (+10.7«</td><td> 8.11 (+2.11«</td>

<td> benzyl alcohol hydrochloride</td><td> 10</td><td> 186 (+18.51)</td><td> 1167 (+11.5«</td><td> 7.89 (+5.1«</td>

<td></td><td> 50</td><td> 175 (+11.51)</td><td> 1391 (+6.9«</td><td> 7.97 (+1.1J)</td>

<td> i-Amino-3,5-dibron1o־a-l (tert־butylamlno)methyll־</td><td> 2</td><td> 161 (+H.5I)</td><td> 1362 (+U.5«</td><td> 8.3 (0.1«</td>

<td> benzyl alcohol hydrochloride</td><td> 10</td><td> 185 (+7.8«</td><td> 1159 (+11.9«</td><td> 7.89 (+5.1«</td>

<td></td><td> 50</td><td> 181 (+17.2«</td><td> 1116 (+8.6«</td><td> 7.70 U7.3«</td>

a

Values given are the total average gain (g) per rat for the entire experimental period.

¹*Values given are the total average feed consumed per rat for the entire experimental period.

Figures in parentheses are J Improvement over control.

40S10/2

EXAMPLE 8 a- [ (Tert-butylamina)methyl]-3,5-dichlorobenzyl Alcohol \ Hydrochloride

A solution containing 35־ g of 3,5-dichlorostyrene 5 oxide in 50 ml of absolute ethanol and 20 ml of t-buty1amine is heated gently at reflux for 8 hours and the mixture is evaporated to dryness. The clear yellow syrup is dissolved in 75 ml of ethanol and 25 ml of H₂O, and the solution is cooled to.5 °C and acidified with 3N HC1. This 10 solution is evaporated to dryness in vacuo and the residual white solid is recr-stallized from acetone to afford 2.81 g, m.p. 218-221°C.

Anal . Called for C,₀H.-NOCKHC1: C, 48.26; H, 6.08; H, 4.69. Found: C, 4849־; H, 6.17; N, 4.66.

The free base of the title compound is obtained by neutralization of the title compound with aqueous 10% NaOH. Other salts of the free base are then obtained by treatment of the free base in the above-mentioned procedure (aqueous ethanol) with addition of the appropriate acids, 20 such as H₂SO₄, H^PO^, HNO^, CH^SO^H, toluenesulfonic acid and pamoic acid־

The intermediate 3,5-dichlorostyrene oxide needed for preparing the title compound is made by reducing 28.4g of 3,5-dichlorophenacyl bromide in 125 ml of absolute ethanol 25 at 5°C with 8g of NaBH^, added portionwise. After the addition is completed, the reaction mixture is stirred 16 hours at ambient temperature, which is obtained by gradual melting of the ice bath overnight. The mixture is quenched with 100 ml of H₂0, the aqueous mixture is cooled to 5°C, 30 and carefully acidified to pH 3 with concentrated HC1. The mixture is extracted with 300 ml of CH₂C1₂ and the extract is dried over MgSO^, filtered, and evaporated to dryness in vacuo to afford the epoxide as a clear yellow oil.

The phenacyl bromide intermediate for the above35 mentioned styrene oxide is prepared by brominating 10 g of

3,5-dichloroac6tophenone in 50 ml of CHCl₃/50 ml of EtOAc with 23.6 g of CuBr₂. The mixture is heated at reflux for

2.5 hours and cooled to room temperature. After stirring for 16 hours at room temperature, the mixture is cooled in ice for 2 hours and filtered. The filter cake is washed with 50 ml of CHC1₃ and the combined filtrates are twice decolorized with activated carbon, filtered, and evaporated to dryness in vacuo to afford the orange oil of the 3,5dichlorostyrene oxide.

EXAMPLE X 9

The following 3,5-dichlorophenyl compounds (A) related to the title compound of Example 8 are prepared by the method described in Example 6 by substituting t-butyl amine with R2R3NH amines.

A \ .-CH-CH -NR_R_ y fa ^{2 2 3}Cl

A

Compound ^R2^3

ΗH ^{2 HCH}^{3 H} CH.Iff ^{4 H}«387־ ⁵ h n-c₃H₇^{6 H} 13^מ08־״ h>

* H cyclohexyl ^{8 H} ch₂-ch=ch₂^{9 H} ch₂-ch=ch-ch₃¹⁰ H CH₂C־CH

6Θ530/2

<td rowspan="2"></td><td rowspan="2"> Compound</td><td colspan="2"> A (Continued)</td>

<td> 1</td><td></td>

<td> e</td><td> 11</td><td> H</td><td> phenyl</td>

<td> כ</td><td> 12</td><td> H</td><td> methoxypropyl</td>

<td></td><td> 13</td><td> H</td><td> benzyl</td>

<td></td><td> 14</td><td> CH״</td><td> CH״</td>

<td></td><td> 15</td><td>^C2^H5</td><td>^C2^H5</td>

<td> 10</td><td> 16</td><td> CH״CH=CH״</td><td> CH״CH־CH״ z z</td>

<td></td><td> 17</td><td> i־^c3^H7</td><td>^1-υ3^Η7</td>

<td></td><td> 18</td><td> CH״-CH=CH״</td><td> -ch₂-ch=ch₂</td>

<td></td><td> 19</td><td> H</td><td> cyclopropyl</td>

<td></td><td> 20</td><td colspan="2"> -ch₂-ch₂-o-ch₂-ch₂-</td>

<td> 15</td><td> 21</td><td> H</td><td> n-butyl</td>

<td></td><td> 22</td><td> H</td><td> C(CH₃)₂-CH₂-&lt; )</td>

<td></td><td></td><td></td><td></td>

<td></td><td></td><td> EXAMPLE .</td><td> 10</td>

a-[(Tert-butylamino)methyl]-3,5-dibromobenzyl. Alcohol Hydro- . 20 chloride

This title compound is prepared from 3,5-dibromostyrene oxide in the same manner as described in Example 9. The starting materials for this styrene oxide are similarly prepared starting with 3’,5^,-dibromoacetophenone.

The corresponding a-[(isopropylamino)methyl]-3,5dibromobenzyl alcohol hydrochloride is prepared by substituting isopropyl amine for t-butyl amine.

EXAMPLE ‘ . 11 m-Hydroxy-a-[(isopropylamino)methyl]benzyl Alcohol Hydrochlo30 ride ^f

In 135 ml of 95% ethanol, 36.75 g of m-hydroxyacetophenone, 36.5 g of benzyl chloride, 1.75 g of KI, and 24.6 g of K₂CO₃ are stirred and heated at reflux for 5 hours. The mixture is cooled, evaporated in vacuo to remove ethanol and 35 100 ml of H₂0 is added. The mixture is then extracted with diethyl ether three times to afford 350 ml of extract, which is further washed with 50 ml of H₂0, saturated NaHCO^ solu «0530/2 tion (2 x 50 ml), 50 ml of H₂0, and 50 ml of brine in succession. The filtrate is dried over Na₂SO₄ and evaporated to dryness. The residual oil is distilled to afford 49.13 g of m-benzyloxyacetophenone, b.p. 145-147®C/0.2 mm. Bromination of 186 g of this acetophenone is accomoplished with 349 g of CuBr₂ in 1£ of CHCL3/I.5 Z of ethanol heated at reflux. A N₂ sweep is used to remove HBr generated. After 4 hours, the mixture is filtered and the filter cake is washed with CHCI3 (2 x 100 ml). The filtrate is evaporated in vacuo to afford an oil, which is dissolved in 200 ml of absolute ethanol (2 x 50 ml), and dried to afford 64.28g m-benzyloxyphenacyl bromide, m.p57-58° ־C. Further cooling of the filtrate affords 34g. A 64 g-sample of the phenacyl bromide is added to a stirred mixture containing 212 ml of i-propylamine in 425 ml of ethanol under N₂ atmosphere at 5°C. The temperature rises to 12 °C and a clear solution is obtained. The solution is poured into ice (2 L) containing 500 ml of concentrated HC1 and 1500 ml of H₂O. After stirring for 20 minutes, the mixture is filtered and the solid is washed with H₂0. On drying this gives 98.64g, m.p. 200-203°€ dec. This solid is dissolved in 400 ml of refluxing methanol, 400 ml of isopropyl alcohol is added, and the solution is concentrated to 400 nl. On cooling and collecting crystals, 54.36 of ketoamine melting at 213215־® dec is obtained. This material (16 g) is added to 150 ml of methanol which contains 2 g of 5% Pd/carbon and hydrogenated in a Paar vessel at 42 p.s.i.g. of H₂. The mixture is filtered and the filtrate is evaporated. The residue is mixed with 50 ml of isopropyl alcohol and evaporated to dryness to afford a syrup, which is mixed with 100 ml of ethanol. They crystals are collected, washed with diethyl ether and dried to give 10.77 g, m.p. 129-132®C, of the title compound.

By substituting tert-butylamine for isopropylamine, m-hydroxy-a-[(tert-butylamino)methyl]benzyl alcohol hydrochloride, m.p. 150-154®C dec. is obtained. Substitution of isopropylamine with diisopropylamine, benzylamine and allyamine affords m-hydroxy-a-[(diisopropylamino)methyl] *05SO/2 benzyl alcohol, m-hydroxy-a-[(benzylamino)methyl]benzyl alcohol, and m-hydroxy-a-[(allylamino)methyl]benzyl alcohol hydrochlorides, respectively.

EXAMPLE 12

4-Amino-a[(tert-butylamino)methyl]-3,5-diiodobenzyl Alcohol HydgQQhi.oyidg

In 10 ml of acetic acid, 0.42 g of ]□-amino-a[(tert-butylamino)methyl]benzyl alcohol is stirred under N₂atmosphere and 0.48g of Ν,Ν-dichlorobenzenesulfonamide and

0.6g of Nal are stirred under N₂ atmosphere for 20 minutes. After 3 days, the mixture is poured into ice and the mixture is basified with 50% aq. NaOH. This mixture is extracted with CH₂C1₂ (3 x 25 ml) and chromatographed on a SiO₂ plate using 1% NH^OH/20% CH^OH/CHjC^ to afford 0.22g of the title 15 compound. The reaction is repeated on a larger scale (8X) and the eluted crude product is dissolved in 100 ml of ethanol/10 ml of H₂0, stirred and 10% HC1 is added to give pH 3. The mixture is evaporated to dryness in vacuo. Isopropyl alcohol is added and the mixture is evaporated to dryness. This process is repeated twice and the residue is crystallized from methanol/isopropyl alcohol by allowing methanol to evaporate until crystals from (methanol is used to dissolve the crude material before isopropyl alcohol is added). On cooling, 2g of the title compound is obtained melting at 187°C dec.

Anal. ¹Calc*d for ^C12^H19^C1I2^N2^O: C,29.02; H,2.86; N,5.64. Found:. C, 29.11; H, 3.64; N, 5.64.

EXAMPLE ' , . 13 a-[(Tert-butylamino)methyl]-3,5-dichlorobenzyl Alcohol Hydro30 chloride

An alternate procedure for preparing the title compound and the compounds described in Example .8-is exemplified. Thus, 10 g of 4-amino-a-[(tert-butylamino)methyl]-3,5dichlorobenzyl alcohol is added to 100 ml of 50-52% H₃PO₂35 and the mixture is stirred and cooled to 8°C in ice while 2.77 g of NaNO₂ in 15 ml of HjO is added over 65 minutes. Foaming occurs and is controlled with antifoaming silicone.

After 20 minutes, the mixture is stirred 2 hours without cooling. The mixture is then poured into ice-H₂0 mixture . and 50% aq. NaOH solution is added until the mixture is alkaline. The alkaline mixture is extracted with CHjClj three times to give 200 ml of solution, which is washed with 25 ml of 2% NaOh and dried over MgSO^ and evaporated to dryness in vacuo to give 9.13 g of brown oil. On standing, the oil solidifies, and it is dissolved in 100 ml of ethanol containing 10 ml of HjO. The solution is acidified to pH 3 with 10% HC1 and evaporated to dryness. The residue is treated with 50 ml of isopropyl alcohol and evaporated to dryness. This procedure is repeated to afford an off-white solid which is dissolved in methanol. The solution is evaporated in vacuo to afford a syrup, which is diluted with 50 ml of isopropyl alcohol and allowed to stand. The crystals which form are collected, washed with isopropyl alcohol and dried to yield 7.8 g, m.p. 217-221°C dec., of the title compound.

The compound described in Example 6 is similarly prepared. Deamination of 4-amino-3,5-dibromo-a-[(tert-butylamino)methyl]benzyl alcohol affords 3,5-dibromo־a~[(tertbutylamino)methyl]benzyl alcohol, m.p. 249-251°C dec.

EXAMPLE . 14

4-Amino-3,5-dichloro-g-methoxyphenethylamine hydrochloride

Under N₂ atmosphere, 11 g of 4-amino-a-[(tert-butylamino)methyl]-3,5-dichlorobenzyl chloride is added to 75 ml of methanol at 0°C. After 20 minutes, the cooling bath is removed and the reaction mixture is stirred at ambient temperature. After the reaction is completed, the mixture is evaporated to dryness in vacuo. The residue is stirred in 75 ml of H₂O and the mixture is made alkaline with 6N NaOH solution and extracted with CH₂C1₂ (3 x 50 ml). The organic phases are dried over MgSO₄ and evaporated to dryness to afford an orange oil. This oil is dissolved in 150 ml of absolute EtOH and acidified with HCl/isopropyl alcohol solution to pH 2. The solution is evaporated to dryness and the residue is stirred in 75 ml of ethyl acetate. After cooling, this affords a pale yellow solid which is 'collected to give _י

6.97 g of the title compound, m.p. 195-198°C dec.

Similarly, substitution of ethyl alcohol, isopropyl alcohol, n-butyl alcohol and n-hexyl alcohol affords the corresponding 0-ethoxy, B-isopropoxy, n-butoxy, and n-hexyloxy phenethylamine hydrochlorides.

EXAMPLE - 15

4-Amino-a-[(tert-butylamino)methyl]-3,5-dichlorobenzyl chloride

Under N₂ atmosphere, 27.72 g of 4-amino-a-[(tertbutylamino)methyl]-3,5-dichlorobenzyl alcohol is added to 200 ml of thionyl chloride stirred at 0-5%C. After addition is completed, the reaction mixture is stirred at ambient temperature for 3 hours. Subsequently, the mixture is evaporated to dryness in vacuo to afford 37.34 g of yellow solid, which is used as is.

EXAMPLE 16

Alternate Procedure for 4-Amino-3,5-dichloro-g-methoxyphenethylamine

I. 100 ml of methanol, 10 g of 4-amino-a-[ (tertbutylamino)methyl]-3,5-dichlorobenzyl alcohol is stirred in an ice bath and dry HC1 gas is introduced into the ,solution. After saturation of the solution, the mixture is stirred at boom temperature for an hour and evaporated to dryness. The solid is then stirred in ethyl acetate to afford the title product, which is collected by filtration.