NZ188912A - Prevention or treatment of copper deficiency by topical application of copper - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number 1 88912 Priori"*/ J ' • f'tfl- 77? b ~l '1 8 rsiOfItjf t— \V|f. « * j i i i y* i i i i i i • • i • i Ccrnplets Specification Filed j r{™ AblK3ll30' fib IK 33)3* 1 ! " 5'T'MAf 19&4' ii - i ww^v. a t 13.5% i i i i • i ■ i « « i ^ ls> ^ i m.i No.: Date: NEW ZEALAND PATENTS ACT, 1953 COMPLETE SPECIFICATION "PROCESS" -+/We, ICI AUSTRALIA LIMITED of 1 Nicholson Street, Melbourne, Victoria, 3001, Australia, a Company organised and existing under the laws of the State of Victoria, Commonwealth of Australia hereby declare the invention for which •§-/ we pray that a patent may be granted to prw/us, and the method by which it is to be performed, to be particularly described in and by the following statement:- - 1 - (followed by page la) 1 8 89"H - la- This invention relates to the prevention or treatment of trace element deficiencies in animals and in particular to the treatment of copper deficiency in warmblooded animals.

Trace elements are required in extremely small amounts for the maintenance of health of both animals and plants. Many soils are deficient in one or more of the essential trace elements and as a result the crops and grasses produced by these soils and the animals feeding on 10 these crops and/or grasses may suffer the same trace element deficiencies. Trace element deficiencies in animals may, depending on the severity of the deficiency, considerably impair the health of the animal.

Various methods have been devised to prevent or 15 treat trace element deficiencies in animals. For example trace elements may be applied to the soil for eventual uptake by the animals, or trace elements may be administered directly to the animal. In general, the application of trace elements to the soil is not an effective means of 20 treating trace element deficiencies in animals as the uptake of the trace elements by crops or grass can vary widely depending on the nature of the crop or grass and the season. Direct administration of trace elements to the animal is the preferred method of treatment. 2<S Certain minerals such as, for example, cobalt and selenium cannot be stored in the body and need to be administered in a form which provides for slow release 183912 of the mineral to the animal. A suitable form for such administration provides the mineral as a heavy pellet which is administered to the animal's reticulum together with a heavy object called a grinder which grinds away any in-5 soluble deposits which may form on the surface of the pellet and allows slow release of the mineral from the pellet.

Copper is a trace element which is essential for the formation of red blood cells, the protective insulating 10 sheath around nerves, certain essential enzymes and for the colour ,of hair and wool. While copper may be administered orally, oral administration may not be effective as it may be immobilized in the rumen by being converted to insoluble complexes. However, copper can be stored in the body and 15 as a result subcutaneous, injection has proved to be the favoured means for its administration.

Unfortunately, the forms of copper used for injection can be toxic and death may follow injection of the normal therapeutic dose if' the copper deficiency is marginal 20 or the system is unable to cope with the sudden increase in the blood copper level.

We have now found that copper deficiency in animals may be prevented or alleviated by the topical application of a suitable formulated copper composition.

The invention provides a process for the prevention or treatment of copper deficiency in a warm-blooded animal excluding humans^which process comprises topically applying to said warm-blooded animal a pharmaceutical composition comprising a copper containing - t 8 39 t 2 compound and a pharmaceutically acceptable carrier effective for passing the copper through the skin of the animal.

It is completely unexpected that copper in a 5 topically applied composition would be effective in the treatment of copper deficiency in animals. However, we ; , have found that the topically applied copper is absorbed into the blood-stream of the animal thereby alleviating the copper deficiency. Our invention thus represents a 10 significant advance in the treatment of copper deficiency . . in animals as it provides-a- facile method of administering^ copper to the animal and allows a gradual uptake of copper by the blood-stream thus overcoming the problems associated with sudden increase in blood copper level resulting from 1£ the administration of copper by subcutaneous infection.' In the process of the invention the copper-^containing pharmaceutical composition is applied directly to the external skin of the animal. The composition may be applied by any of the standard means of topical application 20 such as, for example, spreading on the skin or hide by painting, by brushing or by some other means, spraying onto the skin or hide or simply pouring onto the skin or hide. A preferred means of application is simply pouring a measured quantity of a composition of suitable viscosity 25 onto the skin or hide.

The copper<«-containing pharmaceutical composition may be in the form of a gel, an ointment, a paste or an emulsion such as a cream or lotion but preferably the composition is in the form of a liquid comprising the copper 30 compound either in solution or suspension. Liquid compositions are preferred because of their ease of use, for example in measurement and application, and solutions are P particularly preferred because they are more efficient in transferring the copper compound through the skin. 35 The copper containing compound used in the process • 188912 of the invention is preferably a pharmaceutically acceptable copper salt, copper complex or organocopper compound. In general we have found that ionic copper in association with pharmaceutically acceptable anions in the form of 5 a simple salt and copper in association with pharmaceutically acceptable anions in the form of an anionic complex is particularly suitable for.use in the process of the invention. Suitable anions include pharmaceutically acceptable inorganic anions such as, for example, chloride, bromide, 10 nitrate, sulfate and phosphate and pharmaceutically acceptable organic anions .sjuch as, . for example, acetate, benzoate,, succinate, malate, ascorbate, butyrate, glycinate, lactate, laurate, oleate, naphthenate, palmitate, stearate, tartrate and salicylate. , -15:■ n ^ Pnefe.rably ,the ^pharmaceutical compositions contain copper in the +1 oxidation state and therefore cuprous salts and cuprate (I) complexes are preferred for use in the process of the invention. 3 :i3: 3 ^ : -Where the copper is^rin the form of an anionic com- plex the anionic complex ions are associated with equivalent concentrations of cations. The nature of the associated cations is not critical as long as they are pharma-i i r l h ^li i j .wi.i : oeutiearliy acceptable and-suitable cations are for example 1 the ions of hydrogen, of alkali metals, of alkaline earth 25 metals and ammonium ions. In addition such anionic complexes may optionally contain pharmaceutically acceptable - nonioni& ligands such--a-s sulphur dioxide or nonioni'c7 : in organic compounds.

The carrier used in the process of the invention 30 preferably comprises one or more pharmaceutically acceptable liquids selected from: aromatic hydrocarbons such as, for example, xylenes, toluene and aromatic hydrocarbon fractions of boiling point 130° to 250°C; aliphatic hydrocarbons such as, for example, -to C2Q paraffins, " ' 35 eyeloparaffins "or naphthefies; halogenated aliphiatic Hydro- t 009 1 2 carbons such as, for example, ethylene dichloridej ketones such as, for example, cyclohexanone, methyl ethyl ketone and diethyl ketone; esters such as, for example, lower alkyl salicylates, lower alkyl benzoates, fatty acid esters 5 and glycol and glycerol esters; alcohols such as, for example, short chain (C^ to C^q) and long chain C^ "k° ^20 alcohols including butanol, isopropanol, amyl alcohol and cetyl alcohol; glycols such as, for example, monopropylene glycol;' (poly)alkylene glycols such as, for example, 10 oligomers and polymers prepared from the condensation of . ethylene oxide, propylene oxide- and butylene oxide or - — mixtures thereof and block copolymers comprising blocks of condensed ethylene oxide, propylene oxide and/or butylene oxide; esters of the abovementioned (poly)alkylene -15 glycols such as., for example, the condensation-products of fatty acids with ethylene oxide, propylene oxide, butylene oxide or mixtures thereof; ethers of the above-mentioned (poly)alkylene glycols such as, for example, the .■3 condensation products of^ alcohols and phenols with ethy- lene oxide, propylene oxide, butylene oxide or mixtures thereof; amine alkoxylates such as, for example, the condensation products of ammonia, alkylamines or dialkylamines - with ethylene oxide, propylene oxide, butylene oxide^ r6r"" : J mixtures thereof; ethers and cyclic ethers such as, for example, diisopropyl ether and tetrahydrofuran; amides such as, for example, fatty acid amides and their - 1 1alkoxylates a«d diufethylformamide; sulfones such asj: for •' example, dimethylsulfone or sulfolanej and sulfoxides such as, for example, dimethyl sulfoxide. 30 The specific carrier used will depend to a large extent on the nature of the copper containing compound which comprises the composition and to some extent upon the nature of the skin of the animal which is to be treated by topical application of the composition. As the composition 35 is preferably in the form of a liquid containing the copper 1 889 compound in solution the carrier chosen is preferably a pharmaceutically acceptable solvent for the copper compound.

Preferred carriers include those liquids which are conveniently referred to as (poly)ethers and which may be prepared by the condensation of an alkylene oxide chosen from the group consisting of ethylene oxide, propylene oxide, butylene oxide and mixtures thereof with a compound chosen from the group consisting of water, to alcohols, Cj to Phenols, ammonia, to alkyl- and dialkyl-amines, to carboxylic acids, and mixtures thereof. Suitable (poly)ethers include, for example, pharmaceutically acceptable liquids chosen from: (poly)-alkylene glycols formed by the condensation of ethylene 15 oxide, propylene oxide, butylene oxide or mixtures thereof, or by the sequential condensation of two or more of these alkylene oxides; alcohol alkoxylates formed by alkoxylation of an aliphatic alcohol containing 1 to 24 carbon atoms with ethylene oxide, propylene oxide, butylene oxide or 20 mixtures thereof or by sequential alkoxylation of an aliphatic alcohol containing 1 to 24 carbon atoms with two or more of these alkylene oxides; alkylphenol alkoxylates formed by alkoxylation of an alkylphenol containing 7 to 24 carbon atoms with ethylene oxide, propylene oxide, butylene 25 oxide or mixtures thereof or by sequential alkoxylation of an alkylphenol containing 7 to 24 carbon atoms with two or more..o,f,. these., alkylene,,oxides; amine alkoxylates formed by alkoxylation of ammonia, or a primary or secondary alkylamine containing 1 to 24 carbon atoms, with ethylene 30 oxide, propylene oxide, butylene oxide or mixtures thereof -or by sequential alkoxylation of. ammonia, or a primary or secondary alkylamine containing 1 to 24 carbon atoms, with two or more of these alkylene oxides; fatty acid alkoxylates formed by alkoxylation of a fatty acid containing 1 fco .24"cai^bon):atom&1 with^ ethylene oxide, propylene oxide, ■ -*• 18 3912 butylene oxide or mixtures thereof, by sequential alkoxylation of a fatty acid containing 1 to 24 carbon atoms with two or more of these alkylene oxides, or by the esterification of a fatty acid containing 1 to 24 carbon 5 atoms, or an acid halide or a lower alkyl ester derivative thereof, with fc>aly)alkylene glycol (hereinbefore defined); fatty acid amide alkoxylates formed by alkoxylation of a fatty acid amide containing 1 to 24 carbon atoms with ethylene oxide, propylene oxide, butylene oxide or mix-10 tures thereof, by sequential alkoxylation of a fatty acid .amide containing 1 to -24, carbon atoms with two or more of these alkylene oxides, or by the reaction of a fatty acid containing 1 to 24 carbon atoms, or an acid halide or lower alkyl ester derivative thereof, with a primary . 15, or secondary, amine^alkoxylate obtained by alkoxylation of ammonia or a primary amine; and inorganic ester derivatives, such as for example, sulfate and phosphate ester derivatives, of the above defined (poly)alkylene glycols, : r: alcohol alkoxylates, phenol, alkoxylates, amine alkoxylates, fatty acid alkoxylates or fatty acid - (poly)alkyleneglycol esters and fatty acid amide alkoxylates.

Preferred (poly)ether carriers are the (poly)alkylene , -:,:_ - j ,r; ^ glycols and the alcohol- alkoxylates and in particular : j ---pharmaceutically acceptable condensates of 1 mole; of a 25 Cx to C2q aliphatic alcohol with 1 to 30 moles of an alkylene oxide chosen from ethylene, oxide, propylene oxide l i_u -£— and butylene oxide; pharmaceutically acceptable random or sequential cocondensates of 1 mole of a to aliphatic alcohol with a total of two or more alkylene oxides chosen 30 from ethylene oxide, propylene oxide and butylene oxide; and pharmaceutically acceptable poly(propyleneoxy)-pdly-(ethyleneoxy) glycol block copolymers comprising polypropylene glycol of molecular weight from 1,000 to 3*000 condensed with an amount of ethylene oxide in the range • , 35- from lo to 70% by weightof1 the total block ^copolymer". 18 89 1 ? Typical examples of preferred (poly)ethers are pharmaceutically acceptable (poly)ethers chosen from the n,-. group consisting of methanol condensed with 1 to 3 moles of ethylene oxide; ethanol condensed with 1 to 3 moles of 5 ethylene oxide; butanol condensed with 1 to 4 moles of ethylene oxide; methanol condensed sequentially or randomly --with 2-6 moles of ethylene, oxide and 2 to 6 moles of ^propylene oxide; C^-Cg saturated, branched or linear, aliphatic alcohols condensed with 2-10 moles of ethylene oxide and 10 optionally further condensed with 2-10 moles of propylene oxide or butylene oxide; branched or linear, alcohols condensed with 4-30 moles of ethylene oxide and optionally condensed with 2-20 moles of propylene oxide or butylene oxide; C^-Calkyl phenols, Cg-Cfatty amides, Cg-CgQ 15> -fatty amines, or diphenylol propane, each condensed with 4-20 moles of ethylene oxide and optionally condensed with 4~20 moles of propylene oxide or butylene oxide; polypropylene glycol of molecular weight 900-4000 condensed with 10 to 90% of- the total molecule by weight with : 20 ethylene oxide; and derivatives thereof prepared for example by esterification with inorganic or organic acids.

More preferred (poly)ethers include liquid alcohol alkoxylates chosen from the products of condensation of to aliphatic alcohols with one or more alkylene 25 oxides chosen from ethylene oxide and propylene oxide in the mole ratio of 1:10. Particularly preferred (poly)ethers are the alcohol alkoxylates 2-ethoxy ethanol and the product of sequential condensation of 1 mole of methanol with 4 moles of ethylene oxide and 2 moles of propylene oxide. 30 Preferred carriers also include aliphatic hydro carbon solvents including those hydrocarbon solvents which contain straight chain paraffins, isoparaffins and cyclo-paraffins and mixtures thereof.

In addition to the copper containing compound and 35 the carrier effective for passing the copper through the 1 8 89 1 skin of the animal, the pharmaceutical composition may comprise additives to facilitate its use. For example, the compositions may contain: thickening agents to increase the viscosity of the composition and retard or 5 prevent the composition from running off the animal; surface active agents to improve the contact between the composition and the skin or hide of the animal; stabilizing agents to improve the storage life of the copper containing compound and/or composition as a whole; and 10 anti-inflammatory agents to prevent or alleviate undesirable skin reactions which might be caused by the copper containing compound or carrier.

Surface active agents which may be used in the compositions include pharmaceutically acceptable materials 15 variously called wetting.agents, emulsifying agents and dispersing agents. Suitable surface active agents may be chosen from non-ionic compounds such as, for example, the condensation products of fatty alcohols, alkylphenols, fatty acids, fatty: amineSj fatty acid amides with ethylene t oxide, the fatty acid esters and alkyl ethers of sugars and polyhydric alcohols and their ethoxylate derivatives, and the block copolymers of ethylene oxide and propylene oxide. Many of these surface active agents may act as both surface active agents and carriers in the pharma-25 ceutical compositions. Other suitable surface active' agents may be chosen from ionic compounds such as, for example^ anionic^ compounds including soaps, fatty alcohol sulfate esters, alkyl sulfonates, alkylbenzene sulfonates, alkylnaphthalene sulfonates, alkyl sulfosuccinates 30 and cationic compounds including quaternary alkylammonium salts.

Other additives which may be used to give the composition a suitable hydrophobic balance and aid contact of the pharmaceutical compositions with the skin include 35 animal fatsJ or' waxfes such as, for example lanolin, kineral t I t — i >' oils such as, for example, liquid paraffin or white spirits, vegetable oils such as, for example corn oil, peanut oil,, maize oil, olive oil or castor oil, and polymers such as, for example, polyisobutene.

Compositions suitable for use in the process of the invention include those "copper—in—polyether" compositions disclosed in Australian Patent Application No 11,226/76. In these compositions the copper is believed to be in the form of a sodium chlorocuprate (I) complex, however, the 10 precise nature of the complex is unknown and as a result .this theory should not be regarded as limiting.

The "copper-in-polyether'' compositions disclosed and claimed in New Zealand Patent Specification No. 180100 have been found to be particularly effective in transferring copper 15 through the skin thereby alleviating the copper deficiency. However, preferably such compositions are further formulated to give a composition of suitable viscosity and hydrophile - lipophile balance for use as a pour on.

A preferred composition for use in the process of the invention comprises copper (i) in association with halide ions, a carrier comprising (poly)ether, hydrocarbon solvent-and surface active agent. The composition optionally may contain further additives chosen from 25 (poly)ethers, hydrocarbon solvents, surfactants, mineral oils, vegetable oils and thickening,agents.

Suitable compositions may be prepared by extracting an aqueous solution saturated with cuprous chloride and an alkali or alkaline earth metal chloride with a 30 (poly)ether followed by formulation of the extract with hydrocarbon solvent, surface active agent and further (poly)ether as required.

For example, the extraction of an aqueous solution saturated with both cuprous chloride and sodium chloride 35 with a polyether obtained by the sequential condensation fit rft i •A ^26MAR198l 18 89 1 2 methanol with ethylene oxide and propylene oxide in a mole ratio of 1:4:2, gives a polyether solution containing up to 10% by weight of copper. Such an extract is particularly suitable for use in the preparation of composit-5 ions of the invention as the high concentration of copper allows the formulation of compositions with a lower copper content which are suitable for use as "pour-ons". The copper content, hydrophile - lipophile balance and viscosity of the extract may be adjusted by the addition 10 of hydrocarbon solvents, surface active agents, mineral , , oils, vegetable oils, thickening agents and (poly)ethers > as required.

Preferred compositions comprising copper I in association with chloride ions, a carrier comprising 15 (poly)ether, hydrocarbon-solvent and surface active agent may also be prepared by: a) exposing a mixture of cupric chloride dihydrate in a (poly)ether to visible and/or ultraviolet light; or b) dissolving a mixture of>cuprous chlorine in a liquid 20 comprising a (poly)ether and water; and c) formulating the "copper-in-(poly)ether" solution so obtained in the manner described in the preceding paragraph. 1 In certain of the preferred pharmaceutical com-25 positions used in the process of the invention which comprise copper (I) in (poly)ether it is preferable to maintainthe copper in the +1 oxidation state. This may be done by preventing air from contacting the compositions or, if oxidation has occurred, which may be indicated by 30 a change in the appearance of the solution from colourless to progressively darker shades of yellow and brown, simply by exposing the solution in a clear glass, stoppered container, to sunlight. If preferred, oxidation of the cuprous copper in the compositions may be 35 prevented by the' addition of a pharmaceutically "accept- 1 889 able reducing agent. Suitable reducing agents are sulphur dioxide, sodium metabisulphite, ammonium bisulphite and the like. The reducing agent expressed as equivalent sulphur dioxide can be in the range of 0.05 to 15 percent 5 by weight and is typically from 0.05 to 1.0 per cent by weight. The polyether will act as a reducing agent in the presence of sunlight.

The pH of the pharmaceutical compositions used in the process of the invention may be varied within wide 10 limits according to the type of application. However, in order to minimise discomfort to the animal we have found that a pH in the range from 1 to 9 is satisfactory and 2 to 4 is preferable.

The amounts of the copper compound, carrier and 15 additives employed-injthe pharmaceutical compositions depend to a large extent on the copper compound used and the method of application of the composition. However, as a general rule the composition may comprise 0.01 to 12%, preferably 0.1 to 1%, by weight of copper, 5 to 20 99«95$j preferably 50 to 99 *5%} by weight of the carrier and 0 to 60$, preferably 5 to 50% by weight of additives.

In the process of the invention the pharmaceutical composition is applied topically. Thus the compositions " may be applied in the form of a solution, a suspension, 25 or may be further formulated into gels, creams, pastes and lotions. However, preferably the composition is in the form of a solution suitable for application as a "pour-on".

A solution suitable for use as a "pour-on" may be 30 formulated as hereinbefore described.

A gel may be obtained by adding a gelling agent to a composition as defined hereinabove, and examples of suitable gelling agents are carboxypolymethylene, polyvinylpyrrolidone, polyvinyl_acetate, cellulose deriva-35 tives such as methyl-, ethyl-, hydroxyethyl-, hydroxy- t 8C9 1 2 propylethyl- or sodium carboxymethyl- cellulose, alginates, bentonites and silica.

An ointment may be obtained by dispersing a composition as defined hereinabove in an essentially 5 immiscible organic phase, for example soft paraffin, optionally in the presence of an emulsifying and/or thickening agent, for example sorbitin monostearate.

A paste may be obtained by thickening a composition as defined hereinabove with a solid material such 10 as magnesium stearate, zinc oxide, a silicate or starch.

Emulsions such as creams or lotions may be obtained by mixing a composition as defined hereinabove with a suitable emulsifying system.

The compositions may also contain other pharma-15 ceutically active ingredients which can be absorbed through the skin and promote the health of the animal.

Such ingredients include, for example, anthelmintics such as tetramisole and its laevorotatory isomer and pesticides which -are known to systemically combat ecto-20 parasites sufch as lice, ticks, warble fly and other nuisance flies, and including the known organophosphorous, pyrethrin and pyrethroid pesticides. Conventional additives such as stabilizing agents, pigments and subfet&nces which deter other animals from licking the composition 25 from the skin of the treated animal may also be employed in the compositions.

As previosuly discussed, in the preferred process of the invention the pharmaceutical composition is applied as a pour-on. The dosage rate depends on a 30 number of factors including the efficacy of transfer of the copper through the skin and hence the nature of the copper containing compound, the carrier, the specific formulation employed and the animal being treated. However, in general, suitable dose rates range from 1 mg to 35 1 g of copper per application. For convenience the 1 889 1 2 volume of the composition employed will range from 1 to 100 ml and the percentage of copper in the formulation may be adjusted to enable the desired dose to be applied in a convenient volume of the composition.

The process of the invention may be used for the treatment of copper deficiency in a wide range of warmblooded animals including domestic and farm animals such as pigs, dogs, cats, horses, goats and especially ruminant animals such as bovine domestic animals (i.e. 10 cattle) and sheep. It may also be used to treat laboratory animals such as rats, mice and guinea pigs and to treat humanbeings.

The process of the invention is particularly suitable for the treatment of copper deficiency in cattle 15 where the ease of application and safety is particularly advantageous. In the treatment of cattle we have found compositions comprising 0.1 to 1.0% w/v of copper, and preferably 0.3 to 0.6% w/v of copper, to be particularly suitable. The improved safety of the process of the in-20 vention is illustrated in that at least five times the dose of copper normally given by inj'ection may be applied to the beast without any noticeable adverse affect on its health. t 8 89 | 2 The invention is now illustrated by, but by no means limited to, the following examples.

Example 1 To one litre of a saturated sodium chloride solu-5 tion was added 100 g of purified cuprous chloride powder. To prevent oxidation the resulting mixture was stirred by passage of nitrogen^until, the cuprous chloride was completely dissolved. Sufficient sulphur dioxide was added to the solution to give it a concentration of approximate-10 ly 0.5 g/l of sulphur dioxide. This solution was then extracted with one litre, of a polyether prepared by sequentially condensing one mole of methanol with four moles of ethylene oxide and two moles of propylene oxide. The phases were allowed to separate and the colourless poly-15 ether layer removed and stored in a well stoppered nitrogen filled container to prevent ingress of air.

Example 2 Cuprous chloride was dissolved to the limit of its solubility in a solution saturated with both sodium 20 chloride and sulphur dioxide at room temperature. Samples of this solution were treated with the polyethers of Table I by the following general method, to extract the cuprous copper into the organic polyether.

A sample (20 ml) of the above saturated solution 25 was extracted with the polyether (20 ml) at room temperature. The phases were allowed to separate and the polyether layer removed and stored in a well stoppered nitrogen filled container to prevent the ingress of air. 18 89t 2 TABLE 1 Experiment Polyether 1.

"Ucon11 660 ("Ucon11 is a registered Trade Mark for a polyalkoxylated alcohol). 2.

A sequential condensate of 4 moles of ethylene oxide and 4 moles of propylene oxide with 1 mole of methanol. 3.

A condensate of 11 moles of ethylene oxide with 1 mole of nonylphenol. 4- Polyethylene glycol of average molecular weight 1500.

. A sequential condensate of 4 moles of ethy lene oxide and 3 moles of propylene oxide with 1 mole of methanol. 6.

A sequential condensate of 4 moles of ethylene oxide and 5 moles of propylene oxide with 1 mole of ethanol.

Example 3 Cupric chloride dihydrate (13«5 g) was partially dissolved in a polyether (100 ml) prepared by sequential 5 condensation of 1 mole of methanol with 4 moles of ethylene oxide and 2 moles of propylene oxide. The resulting mixture in a stoppered clear glass bottle was placed in direct sunlight. After exposure to sunlight for about 50 hours with occasional shaking all of the cupric chloride 10 had dissolved and the originally dark-brown solution had lightened progressively to give a colourless clear solution. Polarographic analysis showed that the copper present in the polyether solution was in the cuprous state.

Lower concentration cuprous copper compositions 1 339 t were prepared following a method analogous to that described above, the solution of the cupric chloride and its reduction to cuprous chloride proceeding at a faster rate.

Example 4 Cupric chloride dihydrate (13.4 g) was added to "Teric" PE62 (86.6 g) ("Teric" is a registered Trade Mark) a polypropylene glycol-polyethylene glycol block copolymer comprising a polypropylene glycol block of average molecu-10 lar weight from 1600 to 1900 condensed with an amount of ethyelne oxide from 10 to 30$ by weight of the total molecule. The copper salt slowly dissolved to give a very dark, almost black, solution and the solution was placed in a clear glass bottle in direct sunlight. On exposure 15 to the sunlight the solution progressively lightened in colour finally becoming a clear pale yellow.

Example 5 Example 4 was repeated using "Teric" PE64 a poly-propylene-glycol-polyethylene glycol block copolymer com-20 prising a polypropyleneglycol block of average molecular weight of 1600 to 1900 condensed with an amount of ethylene oxide from 30 to 50% by weight of the total molecule. Again, on exposure to sunlight the initially dark brown -black solution progressively lightened in colour to givev 25 a clear pale yellow coloured solution.

Example 6 "Teric" 12A23 (25 g) an alcohol ethoxylate comprising an alcohol with an average carbon chain length of 12 condensed with an average of 23 moles of ethylene 30 oxide was diluted to 100 ml with hot water. On cooling the mixture an extremely thick gel formed and the gel was broken by the addition of sodium chloride (27 g) to give a viscous fluid.

A little sulphur dioxide was bubbled through the 35 aqueous solution and then cuprous chloride (1.5 s) was I o f> « - .j y added to the solution. The cuprous chloride dissolved and the mixture separated into two phases with some crystallization of sodium chloride from the lower aqueous phase. The upper, pale yellow organic phase was separated 5 and on analysis was found to contain 1.2$ w/v copper. Example 7 Example 6 was repeated with the exception that the amount of sodium chloride added to the "Teric" 12A23 -water gel was reduced to 10 g. The cuprous chloride 10 dissolved readily on addition to the aqueous solution and no phase separation occurred.

Example 8 Cuprous chloride (4*2 g) was dissolved in a solution of sodium chloride (11 g) and sodium metabisulfite 15 (1.2 g) in water (30 ml). The volume of the solution was adjusted to 40 ml by the addition of water and the solution was then extracted with cyclohexanone (10 ml).

On analysis the cyclohexanone extract was found to contain 2.8$ w/w copper.

Example 9 Cuprous chloride (14.0 g) was added to a mixture of sodium chloride (8.3 g)> sodium metabisulfite (3.0 g) and water (12.5 ml). The mixture was then made up to a volume of 100 ml with "Teric" 402 (an alcohol alkoxylate 25 prepared by the sequential condensation of methanol with ethylene oxide and then propylene oxide in the molar ratio of 1:4:2) and the mixture was warmed with stirring until a clear almost colourless solution was obtained. Example 10 Cuprous chloride (11.0 g) was dissolved in a solution of sodium chloride (27 g) and sodium meta-bisulfite (2.5 g) in water (100 ml). The aqueous solution was extracted with "Teric" 402 (30 ml) and the organic layer was separated and clarified by centrifu-35 gation. On analysis the "Teric" 402 was found to con- I 8 89 | tain 8.9$ w/w copper.

Example 11 This Example illustrates the efficacy of the process of the invention in increasing the blood copper levels of 5 animals.

The "copper-in-polyether" solution prepared as described in Example 10 was diluted with "Teric" 402 to obtain a solution containing 0.63$ w/v of copper. The diluted solution (51 ml) was then applied over a 6 hour 10 period to shaved areas on the backs of three rats (i.e. approximately 17 ml applied to each rat. A control group of three rats was similarly treated with "Teric" 402 (55 ml).

Two days after treatment the animals were 15 sacrificed and pooled samples of blood from each treatment group were analysed for copper. The blood from the rats treated with the "copper-in-polyether" composition was found to contain 3*1 ppm of copper whereas the blood from the rats of the control group was found to contain 20 1.8 ppm of copper.

Example 12 This example illustrates the efficacy of the process of the invention in the treatment of copper deficiency in cattle.

The "copper-in-polyether" solutions prepared in Examples 1 and 3 above were analysed for copper content and then diluted with the polyether as necessary to give solutions comprising 0.9$ w/v copper. Four calves whose blood copper level had been measured at seven days, 2 days 30 and just before treatment were then treated with the solutions in the following manner.

Calf 1 was treated by topical application of 27 ml of the 0.9$ w/v copper solution prepared by the dilution of the copper in polyether composition of Example 3• 35 Calf 2 was treated by topical application of 27 ml 1 BB9 of the 0,9% w/v copper solution, prepared by the dilution of the copper in polyether composition of Example 1.

Calf 3 was treated by topical application of 67 ml of the 0.9% w/v copper solution prepared by the di-5 lution of the copper in polyether composition of Example 3.

Calf 4 was treated by topical application of 67 ml of the 0.9% w/v copper solution prepared by the dilution of the copper in polyether composition of Example 1. 10 The blood copper levels of the four calves as measured both before and after treatment with the copper in polyether pour-on preparations is recorded in Table 2 below.

TABLE 2 Calf Blood Copper Level (^g/ml)* Number Day-7 Day-2 Day 0 +6 hr Day 1 Day 7 Day 28 1 0.43 0.53 0.52 0. 66 0.65 0.74 0.74 2 0.40 0.47 0.52 0.65 0.57 0.72 0.80 3 0.15 0.21 0.24 0.41 0.41 0.60 O.48 4 0.30 0.39 0.43 0.58 O.56 1.09 O.87 * 0.5 to 1.5^ug/ml is regarded as the "normal" blood copper level.

Example 13 This Example illustrates the preparation of copper containing compositions suitable for topical application 20 to cattle.

Sodium chloride 55 g was dissolved in water, the volume of the solution was adjusted to 200 ml and sodium metabisulfite (5 g) was added to the solution. Cuprous chloride (21 g) was added and the clear, dark red-brown 25 solution was adjusted in pH to pH 3 by "the addition, of 18S 9 concentrated aqueous sodium hydroxide.

The aqueous solution was extracted into "Teric" 402 (67 ml) and the organic layer was isolated and clarified by centrifugation. The extract was isolated and found 5 to contain equimolar proportions of copper (9*35$ w/w) and sodium (3.24# w/w) and contained 12.9# w/w water.

The "copper in polyether" solution prepared as described above was formulated using the reagents detailed in Table 3 below to give stable solutions A, B and C 10 suitable for use as "pour-ons" in the treatment of copper deficiency in cattle.

TABLE 3 Formulation Percent in Formulation Agent A B C Copper Solution .4 .4 .4 "Teric" 17A2 v/v - - "Arlacel" C - w/v - "Arlamol" E - - w/v White Spirit v/v - - "Clarisol" 315 v/v /v/v Corn Oil v/v v/v v/v 2-Ethoxy-ethanol to 100 vol to 100 vol to 100 vol Final Percentage Copper w/v 0.5 0.5 0.5 "Teric" 17A2 is a Trade name for the product of ethoxy-lation of a 17 carbon aliphatic alcohol with 2 moles of 15 ethylene oxide. ? 809 12 "Arlacel" £ is a Trade name for a sorbitan sesquioleate surfactant.

"Arlamol" E is a Trade name for the product of propoxy-lation of stearic acid with 15 moles of propylene oxide.

"Clarisol" 315 is a Trade name for a hydrocarbon solvent comprising approximately 26% straight chain paraffins, 18% isoparaffins and 5&% cycloparaffins.

"Teric", "Arlacel", "Arlamol" and "Clarisol" are Trade Marks.

When applied to cattle each of the formulations A, Band C showed good penetration and no adverse tissue reaction was noted.

Example 14 This Example illustrates the preparation of a 15 further copper containing composition suitable for topical application.

Oleic acid (56.5 g) was heated to a temperature of l80°C and cupric carbonate (11.3 g) was added portion-wise to the stirred oleic acid and heating and stirring 20 was continued until the reaction had ceased and all water and carbon dioxide appeared to have been expelled.

The reaction product was dissolved in white spirit and the volume adjusted to 125 ml to give a solution containing approximately 5% w/v copper. 25 The "copper in white spirits" solution prepared as described above was formulated using the reagents detailed in Table 4 below to give a composition suitable for use as a "pour-on".

Claims (24)

- 23 -TABLE 4 Formulation Agent Percent in Formulation Copper Solution "Arlamol" E 2-Ethoxyethanol Final Percentage Copper w/v 10 v/v 20 w/v to 100 vol 0.5 When applied to cattle the formulation showed good penetration and no adverse tissue reaction. 9 188912 WHATCLAIM IS: - 24 -

1. A process for the prevention or treatment of copper deficiency in a warm-blooded animal excluding humans which process comprises topically applying to said warm-blooded animal a pharmaceutical composition comprising a copper—containing compound and a pharmaceutically acceptable carrier effective for passing the copper through the skin of the animal.

2. A process according to claim 1 wherein the copper—containing compound is a pharmaceutically acceptable copper salt, copper complex or organocopper compound.

3. A process according to claim 1 or claim 2 wherein the copper<*containing compound is a copper salt.

4. A process according to any one of claims 1 to 3 inclusive wherein the copper containing compound is a chloride, bromide, nitrate, sulfate, phosphate, acetate, h". benzoate, succinate, malate, ascorbate, butyrate, glycinate, r * , a" lactate, laurate, oleate, naphthenate, palmitate, stearate, "A tartrate or salicylate salt of copper.

5. A process according to any one of claims 1 to 4 inclusive wherein the copper-containing compound is cupric oleate.

6. A process according to claim 1 or claim 2 wherein the copper—containing compound is an anionic complex of copper.

7. A process according to any one of claims 1, 2 or 6 wherein the copper containing compound is an anionic halo- copper complex.

8. A process according to any one of claims 1, 2, 6 or 7 wherein the copper—containing compound is a sodium chlorocuprate complex. ,4FEbl984 > 188912 *. - 25 -

9. A process according to any one of claims 1 to 8 inclusive, wherein the copper is in the +1 oxidation state.

10. A process according to any one of claims 1 to 9 inclusive wherein the carrier comprises a liquid selected from the group consisting of: aromatic hydrocarbons; aliphatic hydrocarbons; halogenated aliphatic hydrocarbons; ketones; esters; alcohols; glycols; (poly)alkylene glycols; (poly)alkylene glycol esters; (poly)alkylene glycol ethers; ethers and cyclic ethers; amides; sulfones; sulfoxides; and mixtures thereof.

11. A process according to any one of claims 1 to 10 inclusive wherein the carrier comprises one or more liquid (poly)ethers which may be prepared by the condensation of an alkylene oxide chosen from the group consisting of ethylene oxide, propylene oxide, butylene oxide and mixtures thereof with a compound chosen from the group consisting of water, C^ to C24 alcohols, to C^4 phenols, ammonia, C-^ to C24 alkyl- and dialkyl- amines, to C^4 fatty acids, and mixtures thereof.

12. A process according to any one of claims 1 to 11 inclusive wherein the carrier comprises one or more liquid alcohol alkoxylates chosen from the products of condensation of a C-^ to C^q aliphatic alcohol with one or more alkylene oxides chosen from ethylene oxide and propylene oxide and wherein the mole ratio of aliphatic alcohol to alkylene oxide is in the range from 1:1 to 1:30.

13. A process according to any one of claims 1 to 12 inclusive wherein the carrier comprises one or more liquid alcohol alkoxylates chosen from the product of condensation of to Cg aliphatic alcohols with one or more alkylene oxides chosen from ethylene oxide and propylene oxide and wherein the mole ratio of aliphatic alcohol to alkylene 188912 ( - 26 -

14. A process according to any one of claims 1 to 13 inclusive wherein the carrier comprises a liquid alkoxylate chosen from the group consisting of 2-ethoxyethanol, the product of sequential condensation of methanol with ethylene oxide and propylene oxide wherein the mole ratio of methanol to ethylene oxide to propylene oxide is 1:4:2, and mixtures thereof.

15. A process according to any one of claims 1 to 14 inclusive wherein the carrier comprises an aliphatic hydrocarbon solvent.

16. A process according to any one of claims 1 to 15 inclusive wherein the composition comprises additives chosen from the group consisting of surface active agents, viscosity modifying agents, vegetable oils, mineral oils and mixtures thereof.

17. A process according to any one of claims 1, 2 or 6 to 16 inclusive wherein the composition comprises a sodium chlorocuprate (I) complex, the. product of sequential condensation of methanol with ethylene oxide and propylene oxide in the mole ratio of 1:4:2, 2-ethoxyethanol, an aliphatic hydrocarbon solvent and a surface—active agent.

18. A process according to any one of claims 1 to 17 inclusive wherein the composition comprises from 0.01 to 12% by weight of copper, from 5 to 99.95% by weight of carrier and from 0 to 60% by weight of additives.

19. A process according to any one of claims 1 to 18 inclusive wherein the composition comprises from 0.1 to 1% by weight of copper, from 5 to 50% by weight of additives.

20. A process according to any one of claims 1 to 19 inclusive wherein the composition is applied by pouring onto the back of the animal. 183912 - 27 -

21. A process according to any one of claims 1 to 20 inclusive wherein the animal is a ruminant animal. «*•

22. A process according to any one of claims 1 to 21 inclusive wherein the animal is a bovine animal.

23. A process according to any one of claims 1 to 22 inclusive substantially as described with reference to any one of Examples 11 to 14 inclusive.

24. A process for the prevention or treatment of copper deficiency in warm-blooded anirtals excluding humans substantially as herein described with reference to the Examples. DATED THIS 13^ DAY OF 19 A. J. PARK & SON PER Qs|' QJjuSU* AGENTS FOR THE APPLICANTS