Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
  • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/10—Organic substances
  • A23K20/116—Heterocyclic compounds
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K50/00—Feeding-stuffs specially adapted for particular animals
  • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
  • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
  • C07D209/04—Indoles; Hydrogenated indoles
  • C07D209/08—Indoles; Hydrogenated indoles with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, directly attached to carbon atoms of the hetero ring

Definitions

• An important goal in animal husbandry is to develop biologically active agents which can increase the quantity and improve the quality of meat obtained from livestock animals .
• Increasing the quantity" of food obtained from livestock animals refers to, inter alia, promoting the growth of livestock animals, increasing the efficiency of feed utilized in raising livestock animals and/or enhancing the production of lean body mass in livestock animals.
• Biologically active agents which cause these effects are commonly referred to as “anabolic agents” .
• Improving the quality" of food obtained from livestock animals refers to, inter alia, reducing the quantity of subcutaneous fat in meat while at the same time retaining intramuscular fat.
• Subcutaneous fat commonly referred to as “trim fat”
• intramuscular fat commonly referred to as "marbling” contributes positively to the flavor of meat and maintains a high Quality Grade. Marbling is a therefore desirable quality.
• Biologically active agents which are modestly lipolytic can reduce subcutaneous fat while retaining the intramuscular fat.
• Certain publications have appeared generally disclosing arylpropanolamines such as U.S. Patent 5,013,761 and WO 97/10825.
• Biologically active agents with these properties can be administered to livestock to improve the economics of meat production by increasing the yield of meat (improved Yield Grade) .
• Biologically active agents with these properties can also increase the profitability of meat production by producing meat with a high Quality Grade which, because it is healthier to consume yet retains its flavor, can command high prices from meat packers and consumers .
• One embodiment of the present invention is a compound represented by Structural Formula (I) :
• Rl is a substituted or unsubstituted aryl group, provided, however, that when -X- is -CH- , then Rl is not a substituted or unsubstituted carbazolyl group;
• R2 and R3 are independently -H, a C1-C4 straight chained or branched alkyl group;
• R4 and R5 are independently -H, a C1-C4 straight chained or branched alkyl group or, taken together with the nitrogen atom to which each is bonded, a non-aromatic heterocyclic ring;
• X is -N- or -CH-; and Ring A and Ring B are independently further substituted with zero, one or two substituents .
• Another embodiment of the present invention is a compound represented by Structural Formula (II) :
• Rl in Structural Formula (II) is a substituted or unsubstituted aryl group; and R2-R5 and Rings A-B in Structural Formula (II) are as described above for Structural Formula (I).
• Another embodiment of the present invention is a method of increasing the quantity and improving the quality of meat obtained from a livestock animal.
• the method comprises administering to the animal an effective amount of one or more compounds represented by Structural Formula (I) or (II) or a physiologically acceptable salt of a compound represented by Structural Formula (I) or (II) .
• the compounds of the present invention are strongly anabolic and modestly lipolytic. As a consequence, these compounds can be administered to livestock to increase the quantity of meat obtained from the animals. They also improve the quality of meat by reducing the amount of subcutaneous fat while retaining the intramuscular fat. Thus, the compounds of the present invention can be used to produce greater quantities of meat which is more healthy to consume and retains the normal flavor, i.e., retains marbling and a high quality grade, and can thereby increase the profitability of meat production.
• the compounds of the present invention are intended for the treatment of healthy animals.
• Figure 1 is a graph showing the average daily weight gain after twenty-eight days of cattle treated with: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body weight per day.
• Figure 2 is a graph showing the feed efficiency ratio after twenty-eight days for cattle treated with: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body weight per day.
• Figure 3 is a graph showing the hot carcass weight for cattle treated with the following over a twenty-eight day period: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body- weight per day.
• Figure 4 is a graph showing the adjusted twelfth rib fat thickness of cattle treated with the following over a twenty-eight day period: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body weight per day.
• Figure 5 is a graph showing the average ribeye area of cattle treated with the following over a twenty-eight day period: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body weight per day.
• Figure 6 is a graph showing the carcass soft tissue composition of cattle treated with the following over a twenty-eight day period: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body weight per day.
• Figure 7 is a graph showing the calculated yield grade of meat obtained from cattle treated with the following over a twenty-eight day period: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body weight per day.
• Figure 8 is a graph showing the conformation score of meat obtained from cattle treated with the following over a twenty-eight day period: a) 0.0 mg of Compound 6 per kilogram of body weight per day; 0.125 mg of Compound 6 per kilogram of body weight per day; 0.250 mg of Compound 6 per kilogram of body weight per day; and 0.5 mg of Compound 6 per kilogram of body weight per day.
• the present invention is directed to a compound represented by Structural Formula (I) or (II) . Also included is a method of improving livestock production by administering one or more compounds of the present invention to the livestock.
• Rl is as defined above for Structural Formula (II) .
• Rl is represented by Structural Formula (IV) (V) , (VI) , (VII) or (VIII) :
• Ring C through Ring I are independently substituted or unsubstituted. Preferably, Rings C through Ring I are unsubstituted.
• Aryl groups include carbocyclic aromatic groups such as phenyl, 1-naphthyl, 2-naphthyl, 1-anthracyl and 2-anthracyl, and heteroaryl groups such as N-imidazolyl , 2-imidazolyl, 2-thienyl, 3-thienyl, 2-furanyl, 3-furanyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 2-pyranyl, 3-pyranyl, 3-pyrazolyl, 4-pyrazolyl, 5-pyrazolyl, 2-pyrazinyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2- oxazolyl, 4-oxazolyl and 5-oxazolyl.
• Aryl groups also include fused polycyclic aromatic ring systems in which a carbocyclic aromatic ring or heteroaryl ring is fused to one or more other heteroaryl rings.
• Examples include 1-benzimidazolinyl, 2-benzimidazolonyl, 1-benzimidthioazolinyl , 2-benzimidthioazolonyl, 1- carbazolyl, 2-carbazolyl, 3-carbazolyl, 4-carbazolyl, 3- indazolyl, 4-indazolyl, 5-indazolyl, 6-indazolyl, 2-benzothienyl, 3-benzothienyl, 2-benzofuranyl, 3-benzofuranyl, 2-indolyl, 3-indolyl, 2-quinolinyl, 3-quinolinyl, 2-benzothiazolyl, 2-benzooxazolyl, 2-benzimidazolyl, 2-quinolinyl, 3-quinolinyl, 1-isoquinolinyl,
• aryl group is a group in which one or more carbocyclic aromatic rings and/or heteroaromatic rings are fused to a cycloalkyl or non-aromatic heterocyclic ring.
• Non-aromatic heterocyclic rings are non-aromatic carbocyclic rings which include one, two or three heteroatoms selected from nitrogen, oxygen and sulfur in the ring that will afford a stable structure.
• the ring can be five, six, seven or eight-membered. Examples include 2-tetrahydrofuranyl, 3-tetrahydrofuranyl,
• aliphatic groups include straight chained, branched or cyclic C1-C20 hydrocarbons which are completely saturated or which contain one, two or three units of unsaturation.
• Suitable- substituents on an aliphatic group, aryl group (carbocyclic and heteroaryl) , non-aromatic heterocyclic ring or benzyl group are those which do not significantly reduce the anabolic effects or alter the lipolytic effects of the compound. Examples include -OH, halogen (-Br, -Cl, -I and
• R is C1-C6 alkyl, benzyl or phenyl.
• a substituted aliphatic, substituted aromatic, substituted non-aromatic heterocyclic ring or substituted benzyl group can have one, two or three substituents.
• Physiologically acceptable salts of the compounds disclosed herein, including the compounds represented by Structural Formulas (I) , (II) and (III) and the compounds shown in Table 1, are also included. Salts can be formed from those compounds which comprise acidic functional groups by reacting with a suitable base.
• Such salts include those derived from inorganic bases such as ammonium and alkali and alkaline earth metal hydroxides, carbonates, bicarbonates, and the like, as well as salts derived from basic organic amines such as aliphatic and aromatic amines, aliphatic diamines, hydroxy alkamines, and the like.
• bases useful in preparing the salts of this invention thus include ammonium hydroxide, potassium carbonate, sodium bicarbonate, calcium hydroxide, methylamine, diethylamine, ethylenediamine, cyclohexylamine, ethanolamine and the like. Because of the amine moiety, salts of the compounds disclosed herein can also be prepared by reacting with a suitable acid.
• Acids commonly employed to form such salts include inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid, as well as organic acids such as para-toluenesulfonic, methanesulfonic, oxalic, para-bromophenylsulfonic, carbonic, succinic, citric, benzoic, acetic acid, and related inorganic and organic acids.
• inorganic acids such as hydrochloric, hydrobromic, hydroiodic, sulfuric and phosphoric acid
• organic acids such as para-toluenesulfonic, methanesulfonic, oxalic, para-bromophenylsulfonic, carbonic, succinic, citric, benzoic, acetic acid, and related inorganic and organic acids.
• physiologically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, mono-hydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, 2-butyne-l,4 dioate, 3-hexyne-2, 5-dioate, benzoate, chlorobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, hippurate, ⁇ - hydroxybutyrate, glycolate
• Structural Formulas (IV) -(VIII) indicates the bond by which the phenyl ring of the bicyclic ring system is connected to the 3-oxygen atom of the molecule represented by Structural Formula (I), (II) or (III) .
• the present invention includes solvates of the compounds of Structural Formula I and the physiologically acceptable salts thereof.
• a particular compound of the present invention or a physiologically acceptable salt thereof may form solvates with water or common organic solvents. Such solvates are included within the scope of compounds of the present invention.
• diastereomers exist for the compounds of Structural Formula I and, depending on the substituents, further diastereomers may exist.
• the compounds of the present invention include mixtures of two or more diastereomers as well as each individual stereoisomer .
• ivestock animals are animals raised for food production. Ruminants or "cud-chewing" animals such as cows, bulls, heifers, steers, sheep, buffalo, bison, goats and antelopes are examples of livestock.
• Ruminants or "cud-chewing" animals such as cows, bulls, heifers, steers, sheep, buffalo, bison, goats and antelopes are examples of livestock.
• Other examples of livestock include pigs and avians (i.e. poultry) such as chickens, ducks, turkeys and geese.
• livestock include fish, shellfish and crustaceans raised in aquaculture.
• the method of the present invention is preferably used with ruminants, such as cows, heifers, bulls and steers, and with avians, such as chickens, turkeys and ducks.
• An "effective amount" of a compound of the present invention is the quantity which, when administered to a livestock animal, increases the quantity of meat and/or quality of meat obtained from the animal.
• Increasing the quantity of meat obtained refers to promoting a greater amount of growth in the animal with a treatment compared with the absence of the treatment.
• increasing the quantity of meat obtained refers to promoting formation of lean body mass.
• the formation of lean body mass is promoted, for example, when there is a higher ratio of muscle to fat as a result of a treatment than in the absence of the treatment.
• increasing the quantity of meat obtained refers to improving the efficiency of utilization of food.
• Food utilization is more efficient when there is a greater body weight gain per a given amount of feed consumed by an animal as a result of a treatment than in its absence.
• Increasing the quality of meat refers to an improvement in carcass quality of the animal.
• Improved carcass quality refers, for example, to the formation of less fatty tissue (subcutaneous fat) and/or greater leanness (improved yield grade) , while retaining intramuscular fat (quality grade) .
• improved carcass quality generally results in meat that is more healthy to consume, e.g., is less likely to cause elevated triglyceride and/or cholesterol levels, yet retains flavor.
• a "modestly lipolytic agent” means a compound affording a change in blood non-esterified fatty acid levels of from about 1 to about 500, preferably from about 1 to about 300.
• the effective amount to be administered will vary somewhat depending upon the particular animal species being treated and the particular active ingredient employed, but generally will be from about 0.2 to about 1000 parts per million (ppm: milligrams of compound/kilogram feed) of total daily feed intake. Such amount will provide a dosage of about 0.002 to about 50 mg/kg body weight.
• a preferred embodiment employs about 0.5 to about 200 ppm, and more preferably from about 1 to about 40 ppm.
• the compound when practicing the method in animals such as ruminants or swine, the compound will be added to the daily feed ration at about 1 to 100 parts per million of the daily feed ration.
• the method of the invention is preferably practiced by orally administering an effective amount of a compound of the present invention to a livestock animal .
• Other routes of administration can be employed, for instance in ovo, intranasal (e.g., by intranasal misting device) or subcutaneous, intramuscular or intravenous injection; however, such routes are less practical.
• a compound of the present invention is preferably admixed with suitable carriers or diluents commonly employed in animal husbandry.
• Animal feedstuffs comprising a compound of the present invention are provided as a further embodiment of this invention.
• Typical carriers and diluents commonly employed in such feedstuffs include corn meal, corncob grits, soybean meal, alfalfa meal, rice hulls, soybean mill run, cottonseed oil meal, bone meal, ground corn, corncob meal, wheat middlings, limestone, dicalcium phosphate, sodium chloride, urea, distillers dried grain, vitamin and/or mineral mixes, cane molasses or other liquid carriers and the like.
• Such carriers promote a uniform distribution of the active ingredient, and more typically comprise about 20 to about 98 percent by weight of the feedstuff.
• the preferred method for orally administering the compounds of the present invention is via the daily feed rations
• the compounds can be incorporated into salt blocks and mineral licks, as well as being added directly to link tank formulations or drinking water for convenient oral consumption.
• the compounds can additionally be formulated with polymorphous materials, waxes and the like for long- term controlled release, and administered to animals as a bolus or tablet only as needed to maintain the desired daily payout of active ingredient.
• Compounds can also be administered orally by gavage treatment and/or applied transdermally .
• the compounds of the present invention can be admixed with conventional carriers such as water, propylene glycol, polyethylene glycols, n- methyl pyrrolidone, glycerol formal, corn oil, sesame oil, calcium stearate, polymeric materials and the like.
• conventional carriers such as water, propylene glycol, polyethylene glycols, n- methyl pyrrolidone, glycerol formal, corn oil, sesame oil, calcium stearate, polymeric materials and the like.
• Such formulations can be molded into pellets and administered as an injection or as a slow-release subcutaneous implant, sustained rumen delivery device or intranasal device.
• Such administrations can be made as often as needed to ensure the proper dosing of active ingredient to obtain the desired rate of growth promotion and improvement in leanness and feed efficiency.
• the compounds of the present invention can be prepared by procedures disclosed in WO 97/10825 to Bell et al .
• the amination of the epoxides in Schemes I and II is carried out under conditions known in the art for this type of reaction.
• the epoxide may be combined with the amine in an alcohol, preferably, ethanol at room temperature to the reflux temperature of the reaction mixture.
• the reaction is carried under conditions generally described in Atkins et al . , Tetrahedron Letters 27:2451 (1986) the entire teachings of which are incorporated herein by reference.
• An example of specific conditions for reacting an epoxide with an amine is provided in Example 5.
• the hydrolysis reaction shown in Scheme II can be carried out according to methods known in the art using, for example, polyphosphoric acid, H2O2 and K2CO3 in dimethylsulfoxide, H2O2 and ammonium hydroxide, H2O2 and sodium hydroxide, potassium hydroxide and t-butanol, or water and HCl .
• An example of specific conditions for hydrolyzing a nitrile is provided in Example 6.
• a preferred method of preparing Compound 6 in Table 1 comprises aminating an epoxide represented by the following structural formula
• the individual optically active isomers of the compounds of the present invention may be prepared from their respective optically active precursors by the procedures described above, or by resolving the racemic mixtures. This resolution can be carried out by derivatization with a chiral reagent followed by chromatography or by repeated crystallization. Removal of the chiral auxiliary by standard methods affords substantially optically pure isomers of the compounds of the present invention or their precursors. Further details regarding resolutions can be obtained in Jacques, et al . , Enantiomers , Racemates, and Resolutions, John Wiley & Sons, 1981.
• Powdered K 2 C0 3 (40 grams, 289 mmol, 300 mesh) was added to dimethyl sulfoxide (DMSO) (200 mL) containing H 2 0 (4 mL) under N 2 at room temperature and the mixture was stirred for 30 minutes.
• 4-Hydroxyindole (25.2 grams, 189 mmol) was added to the mixture (slight exotherm to 27 °C) and the mixture was stirred for 10 minutes.
• S) -Glycidyl nosylate (50.0 grams, 193 mmol, 98.5% ee) was added (slight endotherm) .
• the slurry was stirred for 30 minutes at 20-25 °C and for 23 hours at 25-27 °C until the reaction was complete.
• the mixture was diluted with acetone (400 mL) and filtered.
• the cake was washed with acetone (400 mL) and the combined filtrates were concentrated to a volume of ca. 250 mL under vacuum while maintaining the temperature below 35 °C.
• This concentrate was added dropwise, over ca . 2 hours, to deionized H 2 0 (650 mL) held at a temperature of 15-20 °C with an ice/water bath.
• the product slurry was stirred for 1 hour at this temperature and for 30 minutes at 5-10 °C
• the solid was isolated by filtration and washed with cold deionized H 2 0 (150 mL) .
• the solid was dried under vacuum at 35 °C to yield 30.3 grams of product.
• the concentrate was diluted with ethyl acetate (500 mL) , and a solution of acetic acid (14.2 grams, 235 mmol) in ethyl acetate (250 mL) was added to the resulting solution over 30 minutes.
• the resulting slurry was stirred for 2 hours at room temperature.
• the slurry was filtered and the solid was washed with ethyl acetate (2 x 100 mL) .
• the product was vacuum-dried at 65 °C/5 Torr for 24 hours to give 46.5 grams (89.6%) of a white crystalline solid, mp 209-215.9 (dec).
• the Dean-Stark trap was charged with isooctane, and the system was purged with N 2 . The system was then heated to reflux with vigorous stirring and allowed to reflux for 1 hour. The olive green reaction mixture was then cooled to 30 °C over one hour, and the mixture filtered through paper. The filter cake was washed with DMAC (250 mL) and the filtrate stripped at 80 °C for 1.5 hours under house vacuum to yield a thick, dark green oil. The oil was taken up in dichloromethane (580 mL) and washed with deionized water (1160 mL) . The layers were separated and the organic layer washed with more water (250 mL) . The organic layer was then mixed with water (1 L) and the pH was adjusted to approx.
• the cooled mixture was filtered through a Hy-Flo filtering aid (16.5 grams) and the filter cake was washed with isopropanol (43 mL) .
• the filtrate was concentrated to a net weight of 55 grams under full vacuum at 50 °C .
• ethyl acetate 85 mL
• the resultant solution was concentrated under the same conditions to a net weight of 52 grams.
• the concentrate was then taken up in ethyl acetate (230 mL) , and 2.5% wt/vol NaCl solution (150 mL) was added.
• the biphasic system was vigorously stirred and the pH was adjusted to 7.2 with glacial acetic acid.
• the phases were separated and the organic phase was extracted with 2.5% brine (2 x 50 mL) .
• the organic phase was washed sequentially with NaOH/NaCl solution (50 mL, 0.89 grams NaOH) , and water (50 mL) .
• Salt formation The organic phase was concentrated to 40 grams net weight under the conditions noted above.
• the concentrate was diluted with ethyl acetate and stripped to 44.9 grams to dry the solution azeotropically. The solution was then divided into three equal portions and one third of the solution was concentrated to 14 grams (about 6.83 grams of freebase) .
• To the concentrate was added adequate volumes of ethyl acetate (40 mL) and ethanol (18.7 mL) to bring the solution to a ratio of 1/3.5 ethanol to ethyl acetate, taking into account residual solvent in the product concentrate, and a dilution factor of 12.3 mL/gram of product.
• the solution was brought to reflux and the pH was adjusted to 3.5 with a 0.5 N solution of HCl in ethyl acetate (about 30.7 mL) .
• the solution should now have a ratio of approximately 1/4.3 ethanol to ethyl acetate with a dilution factor of approximately 14.5 mL/gram of product.
• the acidic aqueous extract from the workup above was adjusted to pH 12-13 with 1 N NaOH solution in a vigorously •stirred biphasic system with MTBE (80 mL) .
• the layers were separated, and the organic extract was dried over Na 2 S0 4 , filtered, and concentrated to a solid in 80-90% yield.
• Example 7 Preparation of Salts of Compound 6 Hydrochloride Salt
• a solution of HC1 in ethanol (7.04 mL of a 0.6 M solution, 4.22 mmol) was added to a refluxing solution of the Compound 6 freebase (2.0 grams, 4.2 mmol) in ethanol (9.0 mL) .
• the pH was carefully adjusted to 3.0, adding triethylamine or HC1 as required.
• the solution was seeded and allowed to cool to room temperature, at which point it was stirred overnight.
• the slurry was cooled to 0 °C for 2 hours, filtered, and the filtrate washed with 3 mL of cold ethanol.
• Example 9 Preparation of 2- [4- (2-amino-2- methylpropyl ) phenoxy] -3-pyridinecarboxylic acid, ethyl ester, acetic acid salt.
• Ethyl 2- [4- (2-amino-2-methylpropyl) phenoxy] -3- pyridinecarboxylate (10.3 grams, 32.8 mmol) was dissolved in ethyl acetate (52 mL) and heptane (41 mL) and the solution was heated to reflux.
• Acetic acid (1.97 grams, 38.8 mmol) was added, the solution was seeded, and cooled slowly to room temperature.
• the organic phase was washed with brine (2 x 30 mL) and dried over anhydrous MgS0 .
• the drying agent was removed by filtration.
• the MTBE solvent was exchanged with methanol by concentrating the solution using rotary evaporation, redissolving the residue in MeOH, and reconcentrating again. This process was repeated and the residue was dissolved in anhydrous methanol and used directly in the subsequent reactions.
• Ethanol 100 mL, 2B-3 was added and the solvent again removed by rotary evaporation until 26 grams remained. This addition of ethanol and evaporation was repeated 3 more times for the solvent exchange into ethanol and removal of ammonia. After the last evaporation, the contents of the flask weighed 25.0 grams and was taken to be the theoretical 12.5 g of freebase and 12.5 grams of ethanol. Ethyl acetate (87.7 mL) and H 2 0 (1.0 mL) were added and the solution was brought to reflux. Acetic acid (1.73 grams, 28.8 mmol) was added and the solution was seeded. After 1 hour, heating was removed from the white mixture.
• the solution was cooled to 70-75 °C, and a 70-75 °C solution of 4-hydroxybenzoic acid (5.9 grams, 42.3 mmol) in 2B-3 ethanol (20 mL) was added in one portion.
• the homogeneous solution was seeded and stirring continued at 70-75 °C for 1 hour.
• the mixture was cooled to 26 °C and stirred for 1 hour, then cooled to 5 °C and stirred for an additional hour.
• Methyl ethyl ketone (450 mL, MEK) was added and the volume reduced to 60 mL at 40-50 °C and 25 inches Hg. MEK was again added (200 mL) and the volume reduced to 60 mL. At 15-20 °C the contents were brought back to a total volume 300 mL using MEK, filtered using a 20 micron fritted glass filter funnel, then rinsed with MEK to give a total filtrate volume of 310 mL. The solution was heated to 65 °C and a solution of glacial acetic acid (3.75 g, 62.4 mmol) in MEK (15 mL) at 65 °C was added.
• 4-Hydroxyindazole may be prepared according to procedures disclosed in Davies, J. Chem . Soc . 2955:2412 (1955) and H. D. Porter and W. D. Peterson, "Organic Synthesis", Collective Volume III, p.660. The entire teachings of these references are incorporated herein by reference. Specific conditions for preparing 4- hydroxyindazole are provided below.
• Compounds 2 , 4 and 8 were prepared by the techniques described above and analyzed by electro spray ionization mass spectrometry.
• the molecular ion peak for Compound 2 was 476.0 (calculated molecular weight 475.55); Compound 4 was 508.0 (calculated molecular weight 507.61); and Compound 8 was 492.24 (calculated molecular weight 492.01).
• Example 17 Intravenous Administration of the Compounds of the Present Invention to Cattle Intravenous administration of the compounds of the present invention to cattle increases serum non-esterified fatty acids and decreases serum urea.
• angus/angus cross calves both heifers and steers, weighing approximately 282 pounds (128 kg) initially to 788 pounds (357.8 kg) over the course of these studies, were placed in pens at 5 calves per pen. The cattle were acclimated to the pens for at least 1 week prior to initiating the study.
• Calves were fed ad libi tum twice daily, (approximately 6-15 pounds (2.7-6.8 kg) /day) .
• the feeding times were staggered to ensure that all animals were fed approximately one hour before the treatments were administered.
• the cattle were fed immediately after receiving the P.M. injection.
• NEFA non-esterified fatty acid level
• SUN serum urea nitrogen level
• Compound 6 was mixed with ground corn and fed to the steers as a top dress on a portion of their daily feed.
• the CONTROL steers received a similar amount of a ground corn top dress.
• the basal ration was a commercially available feed (19.3% CP, DM basis) . Both feed and water were available ad libi tum.
• the steers were individually housed in 12 ft X 48 ft pens equipped with an individual feed bunk and automatic waterer. On day 28 the steers were weighed and a blood sample collected prior to being shipped for slaughter and subsequent carcass evaluation.
• Live performance parameters were improved with the cattle on the Compound 6 treatments.
• LOW, MED, and HIGH exhibited a 54% to 73% increase in average daily gain (ADG, pounds/day) over CONTROL, specifically 6.31, 7.07, 6.74, and 4.08 pounds/day, respectively (2.86, 3.21, 3.06, and 1.85 kg/day) ; P ⁇ 0.0002; Figure 1.
• Daily dry matter intake did not differ (P>0.47) between the treatments.
• Feed efficiency (pound of feed per pound of gain) was improved (P ⁇ 0.0006) on LOW, MED, HIGH compared to CONTROL (4.36, 4.19, 4.18, and 7.04, respectively; Figure 2.
• Fat thickness at the twelfth rib was not different (P>.78; Figure 4), but the area of the longissi us dorsi muscle at the twelfth rib was 10.7 to 18.9% larger (P ⁇ .0068) with LOW, MED, and HIGH compared to CONTROL (13.5, 14.3, 14.5, 12.2 in 2 , respectively; Figure 5).
• Carcass composition including % fat, % protein, % moisture, and % bone were calculated using the equations reported by Hankins, O.G., and Howe, P.E. 1946. U.S. Department of Agriculture Technical Bulletin No. 926 pp. 1-20 ( Figure 6) .
• the carcass composition did not differ statistically (P>.12) among the treatments for any of the components, but % fat was numerically (-1.2 to -2.7%) less with the Compound 6 treatments than CONTROL.
• Meat quality was investigated using several different measurements on longissimus dorsi strip steaks. There were no treatment differences (P>.5) in color as measured by Hunter a*, b*, and L*, or pH measured after a 14 day postmortem aging time. Subjective color, texture, and firmness measurements of the longissimus dorsi muscle at the twelfth rib approximately 24 hour post-mortem also showed no treatment differences (F-test P>.07).
• feeding Compound 6 to these steers resulted in improved efficiency of gain due to 54 to 73% greater weight gain on the same amount of feed.
• the higher live weight gain was maintained in the carcass as evidenced by heavier hot carcass weights.
• the extra carcass weight was due primarily to an increased amount of muscling.
• the large increase in muscling was observed without any apparent deleterious effects on the quality of the meat as determined by carcass marbling scores, and color, pH, and Warner- Bratzler shear force of strip steaks aged for fourteen days.
• Example 19 Effect of Compound 6 on Weight Gain and Feed Efficiency During the Last 14 Days of the Growing Period in Broiler Chickens
• Treatments were randomly allotted to pens within each block. Each treatment consisted of 6 pens with 10 birds per pen.
• Day-of-age birds were obtained from Pine Manor Hatchery in Goshen, IN. Approximately 40 birds per pen were set at random upon receipt. On day 30 of the growout period, all birds were weighed and the 15 birds closest to the block mean were selected. These birds were allowed to acclimate until day 35 of age. On that day, all remaining birds were weighed again, and the 10 birds/pen closest to the block mean were chosen for the treatment phase. Birds were provided ad libi tum access to feed and water throughout the trial . All birds were fed a 23% crude protein corn-soy ration from day 1 until day 18 of age. The feed was changed to a 20% crude protein corn-soy ration from day 18 to day 49.
• the treatment feed was mixed using the same 20% crude protein basal ration. Treatments were administered in the feed from day 35 through day 49. Feed consumption was calculated over the entire 14-day treatment period. Birds were weighed at trial completion (day 49), and transported to Purdue University, West Lafayette, Indiana 47907, U.S.A. meat laboratory for slaughter and carcass measurements on day 50. Weights of the hot carcass, fat pad, and viscera were taken on all animals.
• Hot carcass, viscera, and fat pad weights were taken on all treatments. Hot carcass weights were increased in both treatments with the experimental compound compared to control .
• Viscera weights decreased with respect to control for both treatments with the experimental compound.
• F/G is pen feed intake / total pen weight
• the testing facility contained two wings, each containing 30 pens. The pens in each wing were divided into 6 blocks of 5 pens. Treatments were randomly allotted within each block. Each treatment consisted of 12 pens with 10 birds per pen.
• Day-of-age birds were obtained from Pine Manor Hatchery in Goshen, IN. Approximately 40 birds per pen were set at random upon receipt. On day 16 of the growout period, all birds were weighed and the 15 birds closest to the block mean were selected. These birds were allowed to acclimate until day 21 of age. On this day, all remaining birds were weighed again, and the 10 birds/pen closest to the block mean were chosen for the treatment phase. Birds were provided ad libi tum access to feed and water throughout the trial. All birds were fed a 23% crude protein corn-soy ration from day 1 until day 18 of age. The feed was changed to a 20% crude protein ration for day 18 to day 49. The treatment feed was mixed using the 20% crude protein basal ration.
• Treatments were administered in the feed from day 21 through day 49. An interim weight was taken on day 35. Feed consumption was calculated over the entire 28-day treatment period. Birds were weighed at trial completion (day 49) , and were transported to Purdue University meat laboratory for slaughter and carcass measurements on day 50. Weights of the hot carcass; fat pad; viscera; bone-in, skin- on breast; and bone-in, skin-on, leg quarter were taken on all animals.
• Feed efficiency when measured as feed/gain, showed a trend for improvement in both treatments over the 28-day treatment period.
• Carcass evaluation of the hot carcass weights showed significant increases over control animals.
• the fat pad (including both abdominal fat and gizzard fat) was stripped from each bird and weighed to determine the effect of these compounds on fat accretion. Compound 6 showed no significant effect at either dose.
• Each of the treatments demonstrated a trend for decreased viscera weights when compared to control.
• the breast muscle was weighed with the bone and skin attached.
• F/G is pen feed intake / total pen weight
• Active Ingredient means a compound of Structural Formula I or a physiologically acceptable salt or solvate thereof .
• the above ingredients are blended to uniformity to provide a dry flowable premix that can be admixed with a typical animal feed ration at a rate to provide about 20 ppm of active ingredient in the final feed ration.
• the premix can be added to the following swine grower ration for convenient oral administration of the Active Ingredient to swine .
• each Kg of premix contains: 50 g. manganese as manganese sulfate; 100 g. zinc as zinc carbonate; 50 g. iron as ferrous sulfate; 5 g. copper as copper oxide; 1.5 g. iodine as potassium iodide and 150 g. maximum and 130 g. minimum calcium as calcium carbonate.
• Each Kg of premix contains: 77,161 IU Vitamin D2 ; 2,205 IU Vitamin E; 411 mg. riboflavin; 1,620 mg. pantothenic acid; 2,205 mg. niacin; 4.4 mg. Vitamin B12 ; 441 mg. Vitamin K; 19,180 mg. choline; 110 mg. folic acid; 165 mg. pyridoxine; 110 mg. thiamine; 22 mg. biotin.
• Each Kg of premix contains 200 mg. of selenium as sodium selenite .
• ⁇ Trace mineral premix contains: 2.5% manganese as manganese oxide, 0.07% iodine as potassium iodide, 0.3% cobalt as cobalt carbonate, 0.5% copper as copper oxide, and 20.0% zinc as zinc sulfate.
• Each pound of vitamin A and D3 , premix contains 2,000,000 USP units Vitamin A and 225,750 USP units Vitamin D3.
• ⁇ Vitamin premix provides 3000 IU of vitamin A, 900 ICU of vitamin D3 , 40 mg of vitamin E, 0.7 mg of vitamin K, 1000 mg of choline, 70 mg of niacin, 4 mg of pantothenic acid, 4 mg of riboflavin, 100 meg of vitamin B 2 > 100 c 9 °f biotin and 125 mg of ethoxyquin per kg of complete feed.
• 2 Trace mineral premix provides 75 mg of manganese, 50 mg of zinc, 25 mg of iron and 1 mg of iodine per kg of complete feed.

Landscapes

• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Life Sciences & Earth Sciences (AREA)
• Polymers & Plastics (AREA)
• Engineering & Computer Science (AREA)
• Food Science & Technology (AREA)
• Zoology (AREA)
• Animal Husbandry (AREA)
• Health & Medical Sciences (AREA)
• Birds (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Diabetes (AREA)
• Pharmacology & Pharmacy (AREA)
• Medicinal Chemistry (AREA)
• Animal Behavior & Ethology (AREA)
• General Health & Medical Sciences (AREA)
• Public Health (AREA)
• Veterinary Medicine (AREA)
• General Chemical & Material Sciences (AREA)
• Obesity (AREA)
• Hematology (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
• Fodder In General (AREA)
• Indole Compounds (AREA)
• Plural Heterocyclic Compounds (AREA)
• Feed For Specific Animals (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Priority Applications (11)

Applications Claiming Priority (2)

Publications (1)

Family

ID=22598982

Family Applications (1)

Country Status (16)

Cited By (3)

Families Citing this family (4)

Citations (4)

• 2000
  • 2000-11-13 PL PL00355669A patent/PL355669A1/xx not_active Application Discontinuation
  • 2000-11-13 CZ CZ20021644A patent/CZ20021644A3/cs unknown
  • 2000-11-13 WO PCT/US2000/031060 patent/WO2001036413A1/en not_active Application Discontinuation
  • 2000-11-13 KR KR1020027006170A patent/KR20020059730A/ko not_active Application Discontinuation
  • 2000-11-13 BR BR0015578-0A patent/BR0015578A/pt not_active Application Discontinuation
  • 2000-11-13 IL IL14957000A patent/IL149570A0/xx unknown
  • 2000-11-13 CA CA002391654A patent/CA2391654A1/en not_active Abandoned
  • 2000-11-13 EA EA200200569A patent/EA200200569A1/ru unknown
  • 2000-11-13 JP JP2001538902A patent/JP2003514815A/ja not_active Withdrawn
  • 2000-11-13 AU AU19174/01A patent/AU1917401A/en not_active Abandoned
  • 2000-11-13 CN CN00815577A patent/CN1390213A/zh active Pending
  • 2000-11-13 MX MXPA02004653A patent/MXPA02004653A/es unknown
  • 2000-11-14 SV SV2000000216A patent/SV2002000216A/es unknown
  • 2000-11-15 CO CO00086799A patent/CO5221077A1/es not_active Application Discontinuation
  • 2000-11-15 PE PE2000001223A patent/PE20010707A1/es not_active Application Discontinuation
• 2002
  • 2002-05-14 NO NO20022290A patent/NO20022290L/no not_active Application Discontinuation

Patent Citations (5)

Also Published As

Similar Documents

Legal Events