GB2118176A - Pharmaceutical compositions - Google Patents

Abstract

Pharmaceutical compositions containing a 3-hydroxypyrid-2-one or 3-hydroxypyrid-4-one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms are replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms, or a salt thereof containing a physiologically acceptable cation, are of value for removing toxic amounts of metals, particularly iron, from the body. All but five of the above pyridones are claimed as novel compounds

Description

SPECIFICATION Pharmaceutical compositions This invention relates to compounds for use in pharmaceutical compositions.

Certain pathological conditions such as thalassaemia, sickle cell anaemia, idiopathic haemochromatosis and aplastic anaemia are treated by regular blood transfusions. It is commonly found that such transfusions lead to a widespread iron overload, which condition can also arise through increased iron absorption by the body in certain other circumstances. Iron overload is most undesirable since, following saturation of the ferritin and transfertin in the body, deposition of iron can occur and many tissues can be adversely affected, particular toxic effects being degenerative changes in the myocardium, liver and endocrine organs. Such iron overload is most often treated by the use of desferrioxamine.However, this compound is an expensive natural product obtained by the culture of Streptomyces and, as it is susceptible to acid hydrolysis, it cannot be given orally to the patient and has to be given by a parenteral route. Since relatively large amounts of desferrioxamine may be required daily over an extended period, these disadvantages are particularly relevant and an extensive amount of research has been directed towards the development of alternative drugs. However, work has been concentrated on three major classes of iron chelating agents or siderophores, namely hydroxamates, ethylenediamine tetra-acetic acid (EDTA) analogues and catechols. The hydroxamates generally suffer from the same defects as desferrioxamine, being expensive and acid labile, whilst the other two classes are ineffective at removing iron from intracellular sites.Moreover, some cathechol derivatives are retained by the liver and spleen and EDTA analogues possess a high affinity for calcium and so are also likely to have associated toxicity problems.

We have accordingly studied the iron chelating ability of a wide range of compounds and have identified a group of compounds as being of particular use for the treatment of conditions involving iron overload.

According to the present invention a pharmaceutical composition comprises a 3-hydroxypyrid-2one or 3-hydroxypyrid-4-one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms are also replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms, or a salt thereof with a physiologically acceptable cation, together with a physiologically acceptable diluent or carrier.

The 3-hydroxypyrid-2 and -4-ones may carry more than one type of aliphatic hydrocarbon group and, in particular, the group attached to the nitrogen atom may be different from any aliphatic hydrocarbon group or groups attached to ring carbon atoms. Groups attached to carbon atoms are, however, more often the same when more than one is present. The aliphatic hydrocarbon groups, whether attached to a nitrogen or a carbon atom, may be cyclic or acyclic, having a branched chain or especially a straight chain in the latter case, and may be unsaturated or especially saturated. Groups of from 1 to 4 carbon atoms and particularly of 1 to 3 carbon atoms are of most interest.Alkyl groups are preferred, for example cyciic groups such a cyclopropyl and especially cyclohexyl but, more particularly preferred are acyciic alkyl groups such as methyl, ethyl, n-propyl and isopropyl. Where the ring carbon atoms are substituted by an aliphatic hydrocarbon group or groups these groups are preferably methyl but in the case of the group substituting the nitrogen atom larger groups may more often be utilised with particular advantage. Substitution of the ring carbon atoms, which is preferably by one rather than two or three aliphatic hydrocarbon groups, is of particular interest in the case of the 3-hydroxypyrid-4ones, for example at the 6- or particularly the 2-position, whilst the 3-hydroxypyrid-2-ones may more often be used without any additional aliphatic hydrocarbon group substitutent on the ring carbon atoms.

Particularly if the ring carbon atoms are substituted by the large aliphatic hydrocarbon groups, however, there may be an advantage in avoiding substitution on a carbon atom alpha to the

system. This system is involved in the complexing with iron and the close proximity of one of the larger aliphatic hydrocarbon groups may lead to steric effects which inhibit complex formation.

The compounds may, if desired, be used in the form of salts thereof containing a physiologically acceptable cation, for example the cation of an alkali metal such as sodium, quaternary ammonium ions or protonated amines such as the cations derived from tris (tris represents 2-amino-2-hydroxymethyl propane 1 ,3-diol). Salt formation may be advantageous in increasing the water solubility of a compound but, in general, the use of the compounds themselves, rather than the salts, is preferred.

Examples of specific compounds which may be used in compositions according to the present invention are shown by the following formulae (I), (II) and (III):--

in which R is an alkyl group, for example methyl, ethyl, n-propyl or isopropyl, and R1 is hydrogen or an alkyl group, for example methyl. Among these compounds and others of use in compositions according to the present invention, the 3-hydroxypyrid-4-ones are of particular interest.

Many of the compounds described above are novel, although some of the compounds of lower molecular weight are known, for example the compound of formula (I) in which R is methyl, the compounds of formula (II) in which R is methyl and R1 is hydrogen or methyl or R is ethyl and R1 is hydrogen, and the compound of formula (Ill) in which both R and R1 are methyl.

The present invention thus also includes as compounds, per se, a 3-hydroxypryid-2-one or 3hydroxypyrid-4-one in which the hydrogen atom. attached to the nitrogen atom is replaced by an aliphatic hydrocarbon group and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms is also replaced by an aliphatic hydrocarbon group, and salts thereof containing a physiologically acceptable caution, but excluding the specific compounds 3-hydroxy-1 -methyl-pyrid-2- one, 3-hydroxy- 1 -methylpyrid-4-one, 1 -ethyl-3-hydroxypyrid-4-one, 3-hydroxy- 1 ,2-dimethylpyrid-4one and 3-hydroxy-l ,6-dimethylpyrid-4-one.

The 3-hydroxy-pyrid-2-one compounds may conveniently be prepared by nucleophilic substitution at the nitrogen atom of the corresponding 2,3-dihydroxypyridine, for example using an organic halide R'X in which R' represents the aliphatic hydrocarbon group present on the nitrogen atom of the desired 3hydroxypyrid-2-one and X represents an iodo group. The 3-hydroxypyrid-4-one compounds may conveniently be prepared similarly or preferably from the more readily accessible corresponding 3hydroxy-4-pyrone.Thus, the 3-hydroxy-4-pyrone may conveniently be converted to the 3-hydroxypyrid4-one through protection of the hydroxy group, for example as an ether group such as a benzyloxy group, reaction of the protected compound with a compound R'NH2, in which R' represents the aliphatic hydrocarbon group present on the nitrogen atom of the desired 3-hydroxypyrid-4-one, in the presence of a base, for example an alkali metal hydroxide such a sodium hydroxide. The protecting group may then be removed. The compounds may be converted to salts formed at the hydroxy group thereof through its conversion to the anion (OH o O-) by reaction with the appropriate base according to standard procedures.

It.will be appreciated that these are not the only routes available to these compounds and that various altematives may be used as will be apparent to those skilled in the art. In general, however, it is preferred that the compounds are isolated iri substantially pure form, i.e. substantially free from bypro-ducts of manufacture.

The compounds may be formulated for use as pharmaceuticals for veterinary or particularly human use by a variety of methods. For instance, they may be applied as an aqueous, oily or emulsified composition incorporating a liquid diluent which most usually will be employed for parenteral administration and therefore will be sterile and pyrogen free. However, it will be appreciated from the foregoing discussion in relation to desferrioxamine that oral administration is to be preferred and the compounds of the present invention may be given by such a route. Although compositions incorporating a liquid diluent may be used for oral administration, it is preferred to use compositions incorporating a solid carrier, for example a conventional solid carrier material such as starch, lactose, dextrin or magnesium stearate.

Other forms of administration than by injection or through the oral route may also be considered in both human and veterinary contexts, for example the use of suppositories for human administration.

Compositions may be formulated in unit dosage form, i.e. in the form of discrete portions each comprising a unit dose, or a multiple or sub-multiple of a unit dose. Whilst the dosage of active compound given will depend on various factors, including the particular compound which is employed in the composition, it may be stated by way of guidance that satisfactory control of the amount of iron present in the human body will often be achieved using a daily dosage of about 0.1 g to5 g, particularly of about 0.5 g to 2 g, veterinary doses being.on a similar g/Kg body weight ratio. However, it will be appreciated that it may be appropriate under certain circumstances to give daily dosages either below or above these levels. Where desired, more than one compound according to the present invention may be administered in the pharmaceutical compositions or, indeed, other active compounds may be included in the composition.

Although 3-hydroxy-1 -methylpyrid-4-one has previously been recognised?as a siderophore, it has never before been appreciated that compounds such as this might be used in a pharmaceutical context, and with real advantage. We have found that the 3-hydroxypyrid-2- and -4sones described above are particularly suited to the removal of iron from patients having an iron overload. The compounds form neutral 3:1 iron complexes at most physiological pH values, and have the advantage that they do not co-ordinate calcium or magnesium. Both the compounds and their complexes will partition into noctanol indicating that they will permeate biological membranes, this property being confirmed in practice by tests of the ability of the 59Fe labelled iron complexes to permeate erythrocytes.The measured coefficients (Kp,,rt) for partition of various of the compounds and their iron complexes are presented in Table 1 of Example 5 hereinafter. Although the ability of both the free compound and its iron complex to permeate membranes is important, it is also desireable for both to possess some degree of water solubility. Preferred compounds show a value of Kpart for the free compound of above 0.05 but less than 3.0, especially of above 0.2 but less than 1.0, together with a value of Kpa,t for the iron complex of above 0.02 but less than 6.0, especially above 0.2 but less than 1.0.Reference to Table 1 will show that the preferences as to the structure of the compounds in compositions according to the present invention which are expressed hereinbefore lead to compounds which have K part values both in the free state and as iron complexes which are broadly in line with the ranges indicated above.

Both the 3-hydroxypyrid-2-ones and the 3-hydroxypyrid-4-ones possess a high affinity for iron (III), as evidenced by log K50 values (log K,,01 is defined as being equal to log J Fe(ljn + 21 - [pK,, + n log a(H+ > + m log aL (Ca++)] where ogss Fe(L)n is the cumulative affinity constant of the ligand in question for iron (III), pK,, is the negative logarithm of the solubility product for Fe(OH)3 and has a value of 39, n and m are the number of hydrogen and calcium ions, respectively, which are bound to the ligand, and a a,(H+} and a, (Ca++) are the affinities of the ligand for hydrogen ions and calcium ions, respectively(.In order to solubilise iron (III) hydroxide, log K,,01 must be greater than 0 and in order to remove iron from transferrin, log ,,o should be in excess of 6.0. The log Kso values for 3-hydroxy-1 -methylpyrid-2-one and 1 ,2-dimethyl-3-hydroxypyrid-4-one, by way of example, are 10.0 and 9.5, respectively, thus comparing favourably with those of the bidentate hydroxamates at about 4.0, of catechols at about 8.0, of desferrioxamine at 6.0, and of diethylenetriamine penta-acetic acid (DTPA) at 2.0. Moreover, the ability of the compounds to remove iron efficiently has been confirmed both by in vitro tests and also by in vivo tests in mice.It is particularly significant that these latter tests are successful whether the compound is given intraperitoneally or orally by stomach tube, the compounds being stable under acidic conditions. Oral activity is not generally present among the other types of compound previously suggested for use as iron co-ordinating drugs and although certain EDTA analogues do show such activity, they possess drawbacks for pharmaceutical use.

Although the major use of the compounds is in the removal or iron, they are also of potential interest for the removal of some other metals present in the body in deleterious amounts. The present invention thus includes the use of a 3-hydroxypyrid-2- or -4-one salt thereof as described above for the removal from the body of toxic amounts of metals, particularly iron. Moreover, the invention also includes a method for the treatment of a patient having toxic amounts of a metal, particularly iron, in the body which comprises administering to said patient an amount of a 3-hydroxypyrid-2- or 4-one or salt thereof as described above to effect a reduction of the levels of this metal in the patient's body.

This invention is illustrated by the following Examples.

EXAMPLES EXAMPLE 1 The preparation of 3-hydroxy-1-methylpyrid-2-one 2,3-dihydroxypyridine (5.55 g) is suspended in methyl iodide (20 ml) in a sealed tube and heated for 24 hours at 1400 C. The reaction is taken to be complete when a dark brown residue forms as a separate phase from the methyl iodide and the tube is then cooled in solid carbon dioxide and opened.

The excess methyl iodide is poured off, distilled water (10 ml) is added to the brown residue, and sulphur dioxide gas is bubbled through the mixture until the aqueous phase becomes clear. The pH of the reaction mixture is adjusted to a value of 6 with 1 M aqueous sodium carbonate and the resulting solution then saturated with ammonium sulphate and extracted with chloroform until the chloroform layer no longer gives a blue coloration when added to ferric chloride solution. The chloroform extracts are combined and dried over sodium sulphate.The solvent is then evaporated under vacuum and the resulting residue is crystallised from petroleum ether (b.p. 1000--1200C) using activated charcoal td, give 3-hydroxy-1-methylpyrid-2-one, m.p. 1 2901 31 OC; Vm,,x (nujol) 1660, 3100 cmw (d6DMSO) 3.6(s,3H), (t,1 6.l(t,lH), 6.8(m,2H), 7.3(s,l H); M+ 125.

EXAMPLE 2 The preparation of other 3-hydroxypyrid-2-ones 2,3-Dihydroxypyridine is reacted with ethyl iodide, n-propyl iodide and isopropyl iodide under' similar conditions to those described in Example 1 for methyl iodide. The reaction mixtures are worked up as described in Example 1 to give the following compounds:- 1 -Ethyl-3-hydroxypyrid-2-one: m.p. 130-1320C; Vmax (nujol) 1620,3100 cm-1; b(d6DMSO) 1.2(t,3H) 3.8(m,2H), 6.0(t,2H), 6.8(m,2H), 8.9(s, 1 H); M+ 139.

3-Hydroxy-1-propylpyrid-2-one: m.p. 1480 C; Vmax (nujol) 1620, 3150 cm-1; (S(d,DMSO) 0.7(t,3H), 1.5(m,2H), 3.7(t,2H), 5.8(t,1 H) 6.5-7.0(m,2H), 8.7(s,1 H); M+ 153.

3-Hydroxy- 1 -(2 '-methylethyl)pyrid-2-one: m.p. 1900 C; L'm,,x (nujol) 1660, 3200 cm-1; b(d6DMSO) 1 .0(d,6H), 6.0(m,1 H), 6.5(t,1 H), 6.7(m,2H); M+ 153.

EXAMPLE 3 The preparation of 3-hydroxy-1,2-dimethylpyrid-4-one 3-Benzyloxy-2-methyl-4-pyrone 3-Hydroxy-2-methyl-4-pyrone (22.2 g) in methanol 225 ml) is added to aqueous sodium hydroxide (25 ml containing 7.5 g NaOH). Benzyl chloride (25.5 g) is added and the mixture is refluxed for 6 hours and is then allowed to cool overnight. The bulk of the methanol is removed under vacuum and the residue is treated with water (50 ml). The mixture is extracted into dichloromethane (3 x 25 ml). The extracts are combined, washed with 5% w/v NaOH (2 x 25 ml), then water (2 x 25 ml) and dried over magnesium sulphate. Evaporation of the solvent gives crude 3-benzyloxy-2-methyl-4pyrone (35 g, 92%) which is purified by distillation in nitrogen under reduced pressure to yield a colourless oil (28 g) of b.p. 1 480C/0.2 mm.

1 ,2-Dimethyl-3-benzyloxypyrid-4-one 3-Benzyloxy-2-methyl-4-pyrone (4.8 g) and methylamine hydrochloride (1.56 g) are dissolved in water (200 ml) and ethanol (100 ml) containing sodium hydroxide (2 g) is added. The mixture is stirred at room temperature for 6 days and is then acidified with concentrated hydrochloric acid to pH 2, and evaporated to dryness. The resulting colourless solid is washed with water and extracted into chloroform (2 x 50 ml). The chloroform extracts are combined, dried over magnesium sulphate, and evaporated to yield 1,2-dimethyl-3-benzyloxypyrid-4-one (3.2 g).

1,2-Dimethyl-3-hydroxypyrid-4-one 1 ,2-Dimethyl-3-benzyloxypyrid-4-one (2 g) is added to concentrated hydrobromic acid (10 ml) and heated in a steam bath for 30 minutes. The resulting mixture is then recrystallised from water to yield 1,2-dimethyl-3-hydroxypyrid-4-one (1 g), m.p. 2300C (with decomposition); Vm,,x (nujol) 1620, 3150 cm~1; G(d,DMSO) 2,3(s,3H), 3.8(s,3H), 6.9(d,1 H), 7.8(d,1 H); M+ 139.

EXAMPLE 4 The preparation of other 3-hydroxypyrid-4-ones 3-Benzyloxy-2-methyl-4-pyrone is prepared as described in Example and is reacted with ethyl amine, n-propylamine, isopropylamine, n-butylamine and n-hexylamine hydrochloride under similar conditions to those described in Example 3 for methylamine hydrochloride. The reaction mixture is worked up and the hydroxy group deprotected as described in Example 3 to give the following compounds: 1-Ethyl-3-hydroxy-2-methylpyrid-4-one: m.p. 1900--1950C; Vmex (nujol) 1620, 3150 cm~1; b(d6DMSO) 1.1 (t,3H), 2.6(s,3H), 3.5(m,2H), 7.3(d,1 H), 8.5(d,1 H); M+ 153.

3-Hydroxy-Z-methyl-1-propylpyrid-4-one: m.p. 1 82 0--1 83 OC; Vm,,x (nujol) 1630, 3200 cm~1; S(d6DMSO) 0.9(t,3H), 1 .6(m,2H), 2.43(s,3H), 4.2(t,2H), 7.1 (d,1 H), 8.1 5(d,1 H); M+ 1 67.

3-Hydroxy-2-methyl-1-(1 '-methylethyl)pyrid-4-one: m,p. 1 98 0--2000C; VmaX (nujol) 1630, 3150 cam~'; b(d6DMSO) 1.28(d,6H),2.43(s,3H),4.8(m,1 H), 7.1 5(d,1 H), 8.1 5(d,1 H); M+ 167.

1-Butyl-3-hydroxy-2-methylpyrid-4-one: m.p. 1 8 8--1 90 0 C: Vmax (nujol) 1630, 3200 cam~1; S(d6DMSO) 0.9(t,3H), 1.3(m,4H), 2.41(s,3H), 4.2(t,2H), 7.2(d,l H), 8.3(d,1 H); M+ 181.

1-Hexyl-3-hydroxy-2-methylpyrid-4-one: m.p. 1660--1680C; Vmax (nujol) 1630, 3200 cm~1; b(d6DMSO) 0.8(t,3H), 1 .3(m,8H), 2.5(s,3H), 4.2(t,2H), 7.4(d,1 H), 8.3(d,1 H); M+ 209.

EXAMPLE 5 Partition data on 3-hydroxpyrid-2-and-4-ones and their iron complexes The partition coefficient Apart, being the ratio (concentration of compound in noctanol)/(concentration of compound in aqueous phase) on partition between n-octanol and aqueous tris hydrochloride (20 mM, pH 7.4), is measured at 200C for various of the compounds of Examples 1 to 4 and for their iron complexes (at 1 0-4M) by spectrophotometry. Acid washed glassware is used throughout and, following mixing of 5 ml of the 10-4M aqueous solution with 5 ml n-octanol for 1 minute, the aqueous n-octanol mixture is centrifuged at 1 ,000 g for 30 seconds. The two resulting phases are separated for a concentration determination by spectrophotometry on each.For the free hydroxypyridones, the range 220-340 nm is used for concentration determinations whilst for the iron complexes, the range of 340-640 nm is used.

Values typical of those obtained are shown in Table 1 where it will be seen that quite small changes in structure such as the replacement of a 1 -propyl group by a 1 -(1 '-methylethyl) group can produce quite large differences in Spar, values.

TABLE 1: Partition coefficients

Partition Coefficient, Kp,,rt Free Iron complex Compound Compound [Fe"'-(compound)3] 3-hydroxy-1 -methylpyrid-2-one 0.44 0.10 1 -ethyl-3-hydroxypyrid-2-one 0.5 1.06 3-hydroxy- 1 -propylpyrid-2-one 0.78 6.20 3-hydroxy-1 -(1 '-methylethyl)-pyrid-2-one 3.10 13.50 3-hydroxy-1 ,2-dimethylpyrid-4-one 0.21 0.05 1 -ethyl-3-hydroxy-2-methylpyrid-4-one 0.40 0.03 3-hydroxy-2-methyl-1 -propylpyrid-4-one 0.67 0.53 3-hydroxy-1 -(1 '-methylethyl)-2-methyl-pyrid-4-one 0.95 0.20 1-butyi-2-hydroxy-2-methylpyrid-4-one 5.30 7.70 EXAMPLE 6 In vitro tests of an iron binding capacity The 3-hydroxypyridones used in this Example were prepared as described in Examples 1 to 4.

(1) Mobilisation of iron from ferritin Horse spleen ferritin (Sigma) was used without further purification and its iron content was estimated spectrophotometrically at 420 nm. The ferritin solution in phosphate buffered saline (Dulbecco-OXOlD, 10-6M, pH H 7.4) was enclosed in a Visking dialysis tube and dialysed against a 3 x 10-3 M buffered solution of one of various pyridones as indicated in Table 2. The absorption spectrum of the resulting iron (III) complex in the dialysis solution was recorded after 6 and 24 hours.

For comparative purposes, the procedure was repeated using a blank control.

The results are shown in Table 2 where the percentage of ferritin-bound iron removed by the compound under test is shown. For comparative purposes, results reported in the literature for similar tests with 1 x 10-3 M desferrioxamine (Crichton et al, J. Inorganic Biochem., 1980, 13, 305) and with 6 x 10-3 M LICAMS (Tufano et al, Biochem. Biophys. Acta, 1981, 668, 420) are also given in the Table.

It will be seen that the pyridone compounds are able to remove iron effectively from ferritin in contrast with desferrioxamine and LICAMS (although the latter will remove iron in the presence of ascorbic acid such a mixture is very difficult to manage clinically). These results shown in Table 2 have been confirmed by separating apoferritin and the 3-hydroxypyridine iron complex from the reaction product in each case by chromatography on Sephadex G 10.

TABLE 2 Removal of iron from ferritin

Percentage of iron removed Compound 6 hours 24 hours Control O 0 3-hydroxy-1-methylpyrid-2-one 11 22 1 -ethyl-3-hydroxypyrid-2-one 14 24 3-hydroxy-1-propylpyrid-2-one 11 21 3-hydroxy-1 -(1 '-methylethyl)-pyrid-2-one 11 20 3-hydroxy-1 ,2-dimethylpyrid-4-one 14 31 1 -ethyl-3-hydroxy-2-methylpyrid-4-one 19 34 3-hydroxy-2-methyl-1 -propylpyrid-4-one 15 26 3-hydroxy-2-methyl-1 -(1 '-methylethyl)-pyrid-4-one 17 24 1 -butyl-2-hydroxy-2-niethylpyrid-4-one 6 7 Desferrioxamine (1 mM) 1.5 LICAMS (6mM) 0 LICAMS (6mM+12mM ascorbic acid) 7 (2) Mobilisation of iron from transferrin Human transferrin (Sigma) was loaded with iron (III) by the method of Bates and Schlaback, J.

Biol. Chem. (1973) 248,3228.591ron (III) transferrin (10-5 M) was incubated with'a 4 x 10-3 M solution in tris HCI(0. 1 M, pH 7.4) of one of various pyridones.as indicated in Table 2 for periods of 4 hours and 1 8 hours. The solution was then dialysed against phosphate buffered saline for 24 hours. The 59Fe remaining in the dialysis tube was then recorded. For comparative purposes, this procedure was repeated with desforrioxamine using incubation for both 4 hours and 1 8 hours and with EDTA using incubation for 4 hours only.

The results are shown in Table 3 in terms of the percentage of transferrin bound iron removed by the compound under test It will be seen that the pyrid-4-one compounds are very effective at iron removal, as compared with desferrioxamine or EDTA, after only 4 hours. Although the efficiency at iron removal of the pyrid-2-one compounds is only at a similar level to that of desferrioxamine and EDTA after 4 hours, it increases markedly after 18 hours whereas the level for desferrioxarnine at 1-8 hours is substantially similar to that at 4 hours.

Similar reactive levels of efficiency were observed when the iron was measured spectrophotometrically. Moreover, the results shown in Table 3 have been confirmed by separating apotransferrin and the 3-hydroxypyridone iron complex from the reaction product in each case by chromatography on Sephadex G 10.

TABLE 3 Removal of iron from transferrin

Percentage of iron removed Compound 6 hours 24 hours Control 0 0 3-hydroxy-1 -methylpyrid-2-one 11 62 1 -ethyl-3-hydroxypyrid-2-one 12 52 3-hydroxy-1-propylpyrid-2-one 15 45 3-hydroxy-1-(1 '-methylethyl)-pyrid-2-one 17 57 3-hydroxy-1 ,2-dimethylpyrid-4-one 90 91 1 -ethyl-3-hydroxy-2-methylpyrid-4-one 88 90 3-hydroxy-2-methyl-1 -propylpyrid-4-one 90 92 3-hydroxy-2-methyl- -(1 '-methylethyl)-pyrid-4-one 94 94 Desferrioxamine 17 22 EDTA 27 - EXAMPLE 7 In vivo tests of iron binding capacity The 3-hydroxypyridones used in this Example were prepared as described in Examples 1,3 and 4.

Mice were injected intraperitoneally with iron dextran (2 mg) at weekly intervals over a four week period. Two weeks after the final injection, the mice were injected via the tail vein with 59Fe lactoferrin (human lactoferrin, 1 mg per injection 2 Ci). The mice were then caged individually. After a ten day period, one of the various pyridones listed in Table 4 was administered to groups of mice at 10 mg per mous either intraperitoneally or intragastrically. The excretion of iron was recorded at either 12 or 24 hourly intervals over a three day period before and a two day period after administration of the compound. For comparative purposes, the procedure was repeated with a blank control and with desferrioxamine, also at 10 mg per mouse.

The results are shown in Table 4, being given on the basis of the control representing 100% excretion, and illustrate the particular advantage of the pyridones as compared with desferrioxamine for oral administration. It should be mentioned that the large standard deviation (SD) values are somewhat misleading as uniformly positive results can yield high SDs which might be taken to suggest that the results are not significantly different from zero. However, this is not the case here, the large SD values being a consequence of the large range among the positive responses (the range of values obtained is given in the Table for each compound).

TABLE 4 Extraction of iron in vivo

Intraperitoneal Intragastric Administration Administration Excretion of Excretion of 59FE+SD 59Fe+SD Number of (Range of values) Number of (Range of values) Compound I Mice percent Mice percent Control 12 100+10 - 3-hydroxy-1 -methylpyrid-2-one 7 150 + 30 3 235 + 30 (107-192) (222-240) 1 -ethyl-3-hydroxy-pyrid-2-one 13 223 + 117 13 188 + 66 (133-590) (95-303) 3-hydroxy-1 -propylpyrid-2-one 13 169 + 49 13 149 + 56 (112-280) (53-260) 3-hydroxy-1 ,2-dimethylpyrid-4-one 7 265 + 70 3 320 + 90 (181--401) (242--425) Desferrioxamine 7 340 + 90 3 90 + 20 (172 472) (80--1 07)

Claims (29)

1. A pharmaceutical composition comprising a 3-hydroxypyrid-2-one or 3-hydroxypyrid-4-one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atmos are also replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms, or a salt thereof containing a physiologically acceptable cation, together with a physiologically acceptable diluent or carrier.

2. A pharmaceutical composition according to Claim 1, in which the or each aliphatic hydrocarbon group is 1 to 4 carbon atoms.

3. A pharmaceutical composition according to Claim 1 or 2, in which the or each aliphatic hydrocarbon group is an alkyl group.

4. A pharmaceutical composition according to Claim 3, in which the or each aliphatic hydrocarbon group is an acyclic alkyl group of 1 to 3 carbon atoms.

5. A pharmaceutical composition according to Claim 1, in which the hydrogen atom attached to the nitrogen atom and, optionally, also one or more of the hydrogen atoms attached to ring carbon atoms are replaced by the same or different substituents selected from the group consisting of methyl, ethyl, n-propyl and isopropyl.

6. A pharmaceutical composition according to any of the preceding claims, in which the pyridone is a 3-hydroxypyrid-2-one substituted only on the nitrogen atom.

7. A pharmaceutical composition according to any of Claims 1 to 5, in which the pyridone is a 3hydroxpyrid-4-one.

8. A pharmaceutical composition according to Claim 7, in which the 3-hydroxypyrid-4-one is substituted on the nitrogen atom and by a single additional substituent at the 2- or 6-position.

9. A pharmaceutical composition according to Claim 8, in which the single additional substituent is a methyl group at the 2-position.

10. A phaarmaceutical composition according to Claim 1, in which the pyridone is 3-hydroxy-1 methyl pyrid-2-one, 1 -ethyl-3-hydroxypyrid-2-one, 3-hydroxy- 1 -propylpyrid-2-one, 3-hydroxy-1 -(1'- methylethyl)-pyrid-2-one, 3-hydroxy- 1 ,2-di methylpyrid-4-one, 1 -ethyl-3-hydroxy-2-methylpyrid-4-one, 3-hydroxy-2-methyl-1 -propylpyrid-4-one or 3-hydroxy-1 -(1 '-methylethyl)-2-methylpyrid-4-one, or a salt thereof containing a physiologically acceptable cation.

11. A pharmaceutical composition according to any of the preceding claims, in which the 3hydroxypyrid-2-one or 3-hydroxypyrid-4-one is in the form of the free compound rather than in the form of a salt thereof.

12. A pharmaceutical composition according to any of the preceding claims, in which the pyridone is in substantially pure form.

13. A pharmaceutical composition according to any of the preceding claims, which contains a physiologically acceptable solid carrier.

14. A pharmaceutical composition according to Claim 1 3 in tablet form.

15. A pharmaceutical composition according to Claim 13 in suppository form.

1 6. A pharmaceutical composition according to any of Claims 1 to 12, which has water or a medium containing an organic solvent as the physiologically acceptable diluent and has the form of a solution, suspension or emulsion.

17. A pharmaceutical composition according to Claim 1 6 in sterile injectable form.

18. A pharmaceutical composition according to any of the preceding claims in unit dosage form.

1 9. A compound being a 3-hydroxypyrid-2-one or 3-hydroxypyrid-4-one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms are also replaced by an aliphatic hydrocarbon group of 1 to 6 carbon atoms, and salts thereof containing a physiologically acceptable cation, but excluding the specific compounds 3-hydroxy-1-methyi-pyrid-2- one, 3-hyd roxy- 1 -methyl-pyrid-4-one, 1 -ethyl-3-hydroxypyrid-4-one, 3-hydroxy- 1 ,2-dimethyl-pyrid-4- one and 3-hydroxy-1 ,6-dimethylpyrid-4-one.

20. A compound according to Claim 19, in which the or each aliphatic hydrocarbon group is of 1 to 4 carbon atoms.

21. A compound according to Claim 19 or 20, in which the or each aliphatic hydrocarbon group is an alkyl group.

22. A compound according to Claim 21, in which the or each aliphatic hydrocarbon group is an acyclic alkyl group of 1 to 3 carbon atoms.

23. A compound according to Claim 19, in which the hydrogen atom attached to the nitrogen atom and, optionally, also one or more of the hydrogen atoms attached to ring carbon atoms are replaced by the same or different substituents selected from the group consisting of methyl, ethyl, npropyl and isopropyl.

24. A compound according to any of Claims 19 to 23, being a 3-hydroxypyrid-2-one substituted only on the nitrogen atom.

25. A compound according to any of Claims 19 to 23, being a 3-hydroxypyrid-4-one.

26. A compound according to Claim 25, in which the 3-hydroxypyrid-4-one is substituted on the nitrogen atom and by a single additional substituent at the 2- or 6-position.

27. A compound according to Claim 26, in which the single additional substituent is a methyl group at the 2-position.

28. A compound according to Claim 19, being 1 -ethyl-3-hydroxypyrid-2-one, 3-hydroxy-1 pyropyl pyrid-2-one, 3-hydroxy- 1 -(1 '-methylethyl)-pyrid-2-one, 3-hydroxy-2-methyl- 1 -propyl pyrid-4one or 3-hydroxy-1 -(1 '-methylethyl)-2-methylpyrid-4-one, or a salt thereof containing a physiologically acceptable cation.

29. A compound being a 3-hydroxypyrid-2-one or 3-hydroxypyrid-4-one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic hydrocarbon group and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms are also replaced by an aliphatic hydrocarbon group, or a salt thereof containing a physiologically acceptable cation, for use in vivo as a chelating agent