GB2136807A

GB2136807A - Pharmaceutical compositions

Info

Publication number: GB2136807A
Application number: GB08407181A
Authority: GB
Inventors: Robert Charles Hider; George Kontoghiorghes; Jack Silver
Original assignee: National Research Development Corp UK
Current assignee: National Research Development Corp UK
Priority date: 1983-03-24
Filing date: 1984-03-20
Publication date: 1984-09-26
Also published as: JPH0672097B2; DE3486441T2; JPH0764815B2; GB8407181D0; SG71089G; JPS59205361A; US4585780A; DE3483973D1; HK37590A; EP0120669A3; USRE35948E; CY1510A; EP0120669A2; GB8308054D0; DE3486441D1; GR81911B; EP0120669B1; JPH0680637A; GB2136807B

Description

1

SPECIFICATION

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

Certain pathological conditions such as thalas saemia sickle cell anaemia, idiopathic haemochroma- 70 tosis and aplastic anaemia aretreated by regular blood transfusions. It is commonlyfound thatsuch transfusions lead to a widespread iron overload, which condition can also arise through increased iron absorption bythe body in certain other circumstances. 75 Iron overload is most undesirable since, following saturation of theferritin and transferrin in the body, deposition of iron can occur and manytissues can be adversely affected, particuiartoxic effects being degenerative changes in the myocardium, liver and endocrine organs. Such iron overload is most often treated bythe use of desferrioxa mine. However, this compound is an expensive natural product obtained bythe 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 exten sive 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 orsiderophores, namely hydroxa mates, ethyl enediam i ne tetra-acetic acid (EDTA) ana logues and catechols. The hydroxamates generally sufferfrom the same defects as desferrioxamine, being expensive and acid labile, whilstthe othertwo classes are ineffective at removing iron from in tracellular sites. Moreover, some catechol derivatives are retained bythe 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 in UK patent application 8308056, published as GB 2,118,176A, we 105 describe a group of compounds which we have identified as being of particular use forthe treatment of conditions involving iron overload. These com pounds consist of 3 - hydroxypyrid - 2 - and - 4 ones in which the nitrogen atom and optionally one or more of 110 the carbon atoms of the ring are substituted by an aliphatic hydrocarbon group, particularly of 1 to 6 carbon atoms. We have nowfound that3 hydroxypyrid - 2 -ones and 3 - hydroxypyrid -4- ones containing other ring substituents are also of particularvalue in thetreatment of such conditions.

According to the present invention a pharmaceutical composition comprises 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 acyl group, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substituted by one or, except in the case of ionisable groups, more than one substi- GB 2 136 807 A 1 tuent selected from aliphaticacy], alkoxy, aliphatic amide,aliphaticamine, carboxy, aliphaticester, halogen, hydroxyand sulpho groups and, optionally, inwhich oneormoreofthe hydrogen atomsattached to ring carbon atoms are replaced by one of said substituents, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substituted by an alkoxy, aliphatic ester, halogen or hydroxy group, but excluding compounds in which said replacement of hydrogen atoms in the compound is effected only by aliphatic hydrocarbon groups, or a saitthereof containing a physiologically acceptable ion or ions, together with a physiologically acceptable diluent or carrier.

The ability of both the free compound and its iron complexto permeate membranes is important in the context of the treatment of iron overload, it is also desirable for both to possess some degree of water solubility. A good indication of the physical properties of a compound and its iron complex in this respect is provided by the value of the partition coefficient (K,, ,) obtained on partition between n - octanol and tris hydrochloride (20 mM, pH 7.4; tris representing 2 amino - 2 hydroxymethylpropane 1, 3 - diol) at 200C and expressed as the ratio (concentration of compound in organic phase)/(concentration of compound in aqueous phase). Preferred compounds show a value of Kp,,t forthe free compound of above 0.02 or 0.05 but less than 3. 0, especially of above 0.2 but less than 1.0, togetherwith a value of Kp., forthe 3:1 hydroxy pyridoneliron(111) complex of above 0.02 but less than 6.0, especially of above 0.2 but less than 1.0.

The following comments upon preferences among the groups used for replacement of hydrogen atoms attached to nitrogen or carbon atoms of the pyridone ring are directed towards the use of compounds having partition coefficients in the free and complexed state which lie in these preferred ranges. For examples of measured partition coefficients of specific cornpounds reference should be made to Table 1 of Example 25.

Compositions of particular interest arethose containing a compound in which the hydrogen atom attached tothe nitrogen atom of the pyridone ring is replaced by a group otherthan an aliphatic hydrocarbon group, although the invention does of course extend to compositions containing a compound in which this hydrogen atom is replaced by an aliphatic hydrocarbon group and a hydrogen atom attached to a carbon atom of the ring is replaced by another type of group. Furthermore, although compounds having a group otherthan an aliphatic hydrocarbon group attached to the ring nitrogen atom may also havethe hydrogen atom of one or more ring carbon atoms replaced bythe same ora different group which is also otherthan an aliphatic hydrocarbon group, it is preferred eitherthatthe ring carbon atoms in such compounds are unsubstituted orthat substitution is limitedto aliphatic hydrocarbon groups.

Morethan one of the ring carbon atoms of the 3 hydroxypyrid - 2 - or-4 one may be substituted, for exampletwo of such atoms, either bythe same Formulae in the printed specification were reproduced from drawings submitted afterthe date of filing, in accordance with Rule 20(14) of the Patents Rules 1982.

substituent group or by different substituent groups, for example by an aliphatic hydrocarbon g rou p and by another type of substituent, although compounds in which none or only one of the ring carbon atoms are substituted, for example by an aliphatic hydrocarbon group, are preferred. Substitution of these atoms is of more interest with the 3 - hydroxypyrid - 4- ones, for example atthe 6 - or particularlythe 2 - position, than with the 3 - hydroxypyrid - 2 - ones. Particularly when the ring carbon atoms are substituted bythe larger groups, however, there may be an advantage in avoiding substitution on a carbon alpha to the -C-C 11 1 0 OH 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.

Where a ring nitrogen or carbon atom is substituted by an aiiphatic hydrocarbon group, this group 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 to6 carbon atoms, particularly& 1 to 4and especially& 1 to 3 carbon atoms, are of most interest. Alkyl groups are preferred, for example cyclicgroups such as cyclopropyl and especially cyclohexyl but, more particularly preferred are acyclic groups such as methyl, ethyl, n - propyl and isop ropyL Where the ring carbon atoms are substituted by an aliphatic hydrocarbon group or groups these are preferably methyl but in the case of a group substituting the nitrogen atom larger groups may more often be utilised with particular advantage.

In the case of substituted alphatic hydrocarbon groups, the preferences as to the nature of the aliphatic hydrocarbon group are broadly as expressed above butthe preferences as to size are somewhat different. Firstly, in the case of such groups attached to nitrogen, groups containing only 1 carbon atom are of less interest, particularly with certain substi- tuent groups such as hydroxy and amine groups, since compounds containing such substituted groups may be difficuitto prepare because of a relative lack of stability of the systems 1 1 -N.C.O- and -N.C.N-.

i 1 1 1 10 i Furthermore, in the case of substituted aliphatic hydrocarbon groups attached both to nitorgen and to carbon atoms, a substituted methyl group containing a hydrophilic substituent may cause the compound to be of too hydrophilic a character unless the effect of this group is balanced bythat of another, more hydrophobic, group. Forthis reason aliphatic hydrocarbon groups of 2 or especially 3 or more carbon atoms are often preferred in this context, particularly wherethe substituent is an amine, carboxy, hydroxy orsulpho group, although groups of 1 or 2 carbon atoms maybe very suitable in certain cases, for example where the su bstituent is an amide group, particularly an N-substituted one. [twill be appreciGB 2 136 807 A 2 ated therefore, that the range of size of aliphatic hydrocarbon groupswhich aresubstituted may conveniently be greaterthan that indicated above as being preferred for unsubstituted groups, for example 1 to 8 carbon atoms, particularly 1 to 6 carbon atoms, and especially 3,4 or 5 carbon atoms when the substituted group contains no additional hydropho- bicgroup.

In general,the circumstances in which the smaller substituted aliphatic hydrocarbon groups are of interest occur eitherwhen the substituent group involved is not of a particularly hydrophilic character orwhen it is of such a character but an Aditionai 9 rou p, such as a la rger al iphatic hyd rocarbon group of, for exam pie, 3 or more carbon atoms is present to provide a balance to the hydrophil ic character. Such a larger grou p may be present in the same substituted a] iphatic hydrocarbons g roup, as is the case with the amide containing g roups referred to above, or separately. In particular, when both the ring nitrogen atom and one or more of the ring ca rbon atoms carry a substituted aliphatic hydrocarbon group, it may be suitable for one of the substituted aliphatic hydrocarbon g roups, for example that carried by the nitorgen atom, to be of a size as discussed above, for example of 3 or 4to 5 or 6 carbon atoms, and forthe other or others to be smaller, for example of 2 or particularly 1 carbon atom. Thus, when the nitrogen atom carries either a substituted aliphatic hydrocarbon group or an acyl group, and also a hydrogen atom or atoms attached to carbon are replaced,then such replacementjor example atthe 6-position, as an alternative or in addition to being effected by an aliphatic hydrocarbon group, may conveniently be effected by a substituted methyl group. Itwill be appreciated that substitution atthe nitrogen atom of the compounds is of particular interest in the present invention and that it is preferred to dictate the hydrophiliclhydrophobic balance in the molecule primarily through such N-substitution. It is more usuallythe case therefore that, when the compounds are C-substituted butthe preferred form of C-substitution involving aliphatic hydrocarbon groups is not present, the C-substituent orsubstituents incorporate substituted aliphatic hydrocarbon groups which are such that preferences as to the size and nature of these groups are broadly as expressed forthe unsubstituted groups, for example being substituted alkyl groups of 1 to 3 carbon atoms and particularly substituted methyl groups such as chloromethyl, ethoxymethyl, and especially hydroxymethyl.

Substituted aliphatic hydrocarbn groups attached tothe nitrogen atom of compounds of use in the present invention maycontain morethan one substituent, for example two substituents of a different type. One example of such a multiply substituted aliphatic hydrocarbon group occurswherethe group is substituted both by a sulphonic orparticularly a carboxylic acid ester group and bya sulphonic or particularly a carboxylic acid amide group. One potential drawbackwith such multiply substituted groups, however, is their increased size and it is the case, therefore, that groups containing one substituent, often terminally substituted on the aliphatic hydrocarbon group, are more commonly employed 0 1 4 3 GB 2 136 807 A 3 and, as indicated herein before, substitution by more than one ionisable group (amine, carboxyorsulpho) is specifically excluded (substitution by morethan one group,onlyone of which is ionisable, is not 5 excluded although not of great interest).

As regards the substituent groups present inthe compounds of use in the present invention, an aliphatic acyl group may contain a sulphonyl or carbonyl group. The lattertype are however preferred and although the acyl group may be a formyi group, alkylcarbonyl groups are of most interest. Such acyl groups may, for example, be of 2 to 4 or 5 carbon atoms, and particularly may contain alkyi groups of the type described above as being preferred as an aliphatic hydrocarbon group substituent at a ring nitrogen orcarbon atom being, for example, -COCH2CH3 or especially -COCH3. Alkoxy groups may conveniently be of 1 to 4 carbon atoms and contain similar aikyl groups to those which are preferred in the a] kylcarbonyl g rou ps, examples of such substituents being ethoxy and pa rticularly methoxy.

Amine su bstituents may consisjof a g rou p - NH2 or its charged equivalent, a g roup - NH3 which will be associated with a physiologically acceptable anion, for exam pie a ch loride or other halide ion, a solu bilising ion such as that f rom methane su lphonic or iseth ionic acid, or an anion derived from the hydroxy group of the ring (OH -> 0-), or such a - NH2 30 or - N3 group in which one or more of the hydrogen atoms is replaced by an alphatic hydrocarbon group, forexample an alkyl group such as is described above as a substituentat a ring nitrogen orcarbon atom. Amide substituents maycontain a sulphonyl or a carbonyl group. The lattertype are, however, of most interest andthefurther discussion will therefore refer to them although it applies equallyto the sulphonyl type. The amide substituent may be of the unsubstituted form -CONH2 or may contain a nitrogen atom which is mono- or di-substituted as just described for the amine substituents, for example being a group -CONI-ICH3, etc. Alternatively, the 1 -CO. N- grouping of the amide substituent may be arranged in the opposite sense so that in an N-su bstituent, for example, the nitrogen atom of the amide grouping is linked to the aliphatic hydrocarbon group which is attached to the nitrogen atom of the ring, the carbonyl g rou p bei n 9 attach ed to a n a 1 i p h atic hyd roca rbo n group, for example an alkyl group such as is described a bove i s a su bstitu e nt at a ri n g n it rog e n o r carbon atom, or in the case of a carboxylic acid amide but not in that of a sulphonic acid amide, to hydrogen.

In the case of an amide group arranged in this opposite sense, the nitrogen atom may carry a hydrogen atom or be mono-substituted as discussed 120 foramide substituents of thefirst mentionedform, thatform of amide substituent being the one of particular interest.

Carboxy and sulpho substituents may be present as the group -C02H or _S03H, or as the anion derived therefrom in combination with a physiologi cally acceptable cation, for example the cation of an alkali metal such as sodium, quaternary ammonium ions or protonate amines such as the cation derived from tris (tris represents 2 - amino - 2 - hydroxymethyl propane 1, 3 - diol). Ester substituents may contain a sulphonyloxy or preferably a carbonyloxy group and may be arranged in either sense, i.e. with a carboxylic acid esterthe group -CO.O- may have eitherthe carbonyl group orthe oxy group linked to the carbon atom of the ring orthe nitrogen atom of the ring (through an aliphatic hydrocarbon group on which the ester group is substituted, where appropriate). The other group of oxy and carbonyl will be linked to an aliphatic hydrocarbon group forming part of the ester group or, in the case wherethis is a carbonyl group may alternatively be linked to hydrogen (this possibility does not apply in the case of sulphonic acid esters). Once again, preferred aliphatic hydrocar- bon groups contained bythe ester group are those described above as substituents in relation to substitution on a ring nitrogen or carbon atom. In the caseof N-substituents, where the ester group is attachedtoan aliphatic hydrocarbon group which is in turn attached to the ring, ester groups in which the carbonylorsulphonyl group is linked to this aliphatic hydrocarbon group are preferred, for example the groups -CH2CO2CH3 and -CH2CO2C2H5. Inthecase of C-substituents the reverse is true and with any C-substituent consisting of an aliphatic hydrocarbon group substituted by an ester group there is a strong preference for the oxy group to be attached to the aliphatic hydrocarbon group which is in turn attached tothe ring, for example as in the groups -CH20.COCH3 and -CH20.COC^. Halogen substituents may conveniently be iodo, fluoro, bromo or especially chloro.

It should be noted that sulphonic acid groups and the corresponding ester and amide groups are of less interest as N-substituents in the case of the pyrid - 2 ones than in that of the pyrid - 4 - ones but that N-substituents consisting of an aliphatic hydrocarbon group substituent by an acyl group are of most interest in the case of the pyrid - 2 - ones.

Among the substituents on the ring nitrogen atom of some particular interest are aliphatic acyl groups and aliphatic hydrocarbon goups substituted by amine, amide, carboxy, aliphatic ester and hydroxy groups, for example the groups -COR1, -(CH2)n-COXR2 and -(CH2L-XH, and to a lesser extentthe more complex type of group -WHAnCH(COY)NHCOR2, in which R, is an alkyl group, for example methyl, ethyl or n - propyi, R2 is hydrogen oran alkyl group, for example methyl, ethyl, n - propyi, isopropyl or butyl, orX is an oxyor imino group,Yis OR, or NR2, n isan integerfrom 1 to 4or6, particularly2,3 or4and m is an integerfrom 2 to 4 or6 particularly 3,4or 5. ftwill be appreciatedthat there isan inter-relation betweenthe preferred values ofn and R2sothatthe groups havingthe higher values ofntendto havethe lowervaluesofR2, and vice versa, so that in groups -(CHAn-COXR2Jor example, -(CHAn- and R2 may conveniently togethercontain 3to 7 carbon atoms, especially4to 6 carbon atoms. Also of interest as N-substitutents in the case where a ring carbon atom is substituted by other than an aliphatic hydrocarbon group are 4 GB 2 136 807 A 4 aliphatic hydrocarbon groups, for example groups R,, R, being as described above.

Among substituents on the ring carbon atoms which are of some particular intest are aliphatic hydrocarbon groups and such groups substituted by a halogen, alkoxy orespeciallya hydroxy group, for examplethe groups IR,, -CH2C1,-CH20C21- 115 and -CH20H in which R, is as described above.

The N-substituents described above may be pre- sentonthe nitrogen atoms of various 3 - hydroxypyrid - 2 - ones and 3- hydroxypyrid -4- ones, in particularly3 - hydroxypyrid - 2 - one, 3 - hydroxypyrid -4- one, 2-alkyl (e.g. methyl and ethyl) - 3 hydroxypyrid -4- ones, 6 -alkyl - (e.g. methyl) - 3 - hydroxypyrid -4- ones, 2,6 - dialkyl (e.g. dimethyl) - 3 hydroxypyrid - 4- ones and 3 - hydroxy- 6 hydroxymethyl - pyrid - 4- one. In the case of the 3 hydroxypyrid - 2 - ones, N-substituent groups of the aliphatic acyl, and amide- and ester-substituted aliphatic hydrocarbon grouptype are of special interestwhilst in the case of the 3 hydroxypyrid -4 ones, N-substituent groups of the aiiphatic acyl and amide-, ester- and hydroxy-substituted aliphatic hydrocarbon group type are of especial interest.

As indicated above, the compounds may contain substituent groups, particularly an aliphatic amine, carboxy or sulpho group, in the saitform. Additionally,the compounds may, if desired, be used in the form of salts formed atthe hydroxy groupthereof through its conversion to the anion (OH --> 0-) and containing a physiologically acceptable cation, for examplethe cation of an alkali metal such as sodium, quaternary ammonium ions or protonated amines such asthe cation 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 ratherthan their salts, is preferred.

Specific examples of N-substituted 3 - hydroxypyr id - 2 - and -4 - ones lacking C-substitution otherthan by an aliphatic hydrocarbon group arethefollowing compounds, with thevarious symbols being as defined above and R being hydrogen, ethyl or especially methyl, whilst p represents 0, 1, 2,3 or4 (the case of R2=H being of less interestwhen p=O). Compounds (IV) in which X is an oxy group and compounds (11), and also (V), in which it is an imino group may be mentioned particularly.

4 3 OH 'Zj 6 In, 2 N 0 1 (CH 2)p 1 C 0 R2 (I) (N:( R 1 (ell 1 Xff (IV) 11 01 C "R (LR 2)n 1 C', dl-:-, X (V) oR R (CH2)n X 2 (Vii) "2 d, R 1 (Cii,j 1 - 1 Xk, (V1) oH (:N:( R 1 (ell 2).

1 X R, V111) Examples of pyridones containing C-substituents otherthan aliphatic hydrocarbon groups and Nsubstituents which are aliphatic hydrocarbon groups are the 1 - alkyl - 6 - halomethyl - 3 - hydroxypyrid 4 ones and the 1 - alkyl - 3 - hydroxy- 6 - hydroxymethy]- pyrid -4 - ones and their 6 - alkoxymethyl analogues, specific examples of which arethe following compounds in which the various symbols are as defined above, the two symbols R, representing thesame or a different alkyl group in (Xl), and Hal represents a halogengroup.

0 (4 3 OR 6 1 1 t N R HalCH 2 (1x) a11z 1 l 011 N 0 1 C 0 XR 2 (11) 0 5 4Y3 R 1 J N R 1 0 /,/ R 80 (11i) it 0 0 OR Oil Roe (X) (n) Although both types of compound are of interest in the context of the present invention, the 3 - hydroxypyrid - 4- ones are of particular interest, whilstthe 3 hydroxyprid - 2 - ones are the compounds of particular interestwhen the compounds described herein are used in theform of their iron complexes for the treatment of iron deficiency anaemia as described in another UK patent application of even date herewith.

3 - Hydroxy - 6 - hydroxymethyl - 1 - methylpyrid - 4one, 1 - (2'- amino ethyl) - 3 - hydroxypyrid - 4 - one, 3 -hydroxy-1 -(2'-hydroxyethyi)-2methylpyrid-4one,l -carboxymethy]-3-hydroxy-2-methylpyrid4-oneandl ethoxycarbonyimethy]-3-hydroxy-2methyl - pyrid -4- one are known compounds but all of the other compounds described above are believed to be novel. The present invention thus also includes as compounds, perse, 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 acyl group, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substituted by one or, except in the case of ionisable groups, morethan one substituent selected from aliphaticacyl,aikoxy,aliphaticamine,aliphatic amide, carboxy, aliphatic ester, halogen, hydroxy 4 and sulpho groups and, optionally in which one or more of the hydrogen atoms attached to ring carbon atoms are replaced by one of said substituents, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substituted by an alkoxy, alipha tic ester, halogen or hydroxy group, but excluding compounds in which said replacement of hydrogen atoms in the compound is effected only by aliphatic hydrocarbon groups, or a saitthereof with a physiolo jo gically acceptable ion or ions, excluding the specific compounds 3 - hydroxy - 6 hydroxymethyl - 1 - methylpyrid - 4- one, 1 - (2'- aminoethyl) - 3 hydroxypyrid - 4 - one, 3 - hydroxy - 1 - (2' hydroxyethyl) - 2 - methyl pyrid - 4 - one, 1 - carboxymethyl -3 - hydroxy-2- methylpyrid - 4- one 80 and 1 -ethoxy-carbonyi-3-hydroxy-2-methylpyrid -4-one.

The 3 - hydroxy - pyrid - 2 - one compounds may conveniently be prepared by nucleophilic substitu tion atthe nitrogen atom of the corresponding 2,3 dihydroxypyridine or one containing groups con vertible to C-substituents present in the desired pyridone, for example using an organic halide WX in which Wrepresentsthe group present on the nitrogen atom of the desired 3 hydroxypyrid - 2 - one 90 or a group convertible thereto and X represents a halogen group. In particular, X mayfor example represent an iodo group when Wrepresents an aliphatic hydrocarbon group or such a group of 2 or more carbon atoms substituted by an aliphatic acyl group such as CH3CO(CHA2-, or a bromo group when Wrepresents an aliphatic, acyl group, such as CH3CO- or CH3CH2C0-, an aliphatic hydrocarbon group substituted by an aliphatic acyl group, such as CH3COCH2-, or an aliphatic ester group, such as C21-1r,0COCH2_. A g roup of the last mentioned type, once introduced into the nitrogen atom, may be hydrolysed to yield an aliphatic hydrocarbon g roup su bstituted by a carboxy (or sulpho) grou p, which carboxy (orsulpho) group may in turn be converted to an amide group or even another ester group.

The 3 - hydroxypyrid - 4 - one compounds may conveniently be prepared similarly or preferablyfrom the more readily accessible corresponding 3 hyd roxy -4 - pyrone or a 3 - hydroxy - 4 - pyrone containing groups convertible to the C-substituent present in the desired pyridone, for example bythe reaction -CH20H ---> -CH2Hal. Thus, the 3 - hydroxy - 4pyrone may conveniently be converted to the 3 - hydroxypyrid -4 - one through protection of the hydroxy groupsjor example as an ether group such as a benzy[oxy group, reaction of the protected compound with a compound R'NH2, in which R' represents the group present on the nitrogen atom of the desired 3 - hydroxypyrid - 4- one or a group convertible thereto, in the presence of a base, for example an alkali metal hydroxide such as sodium hydroxide. The hydroxy protecting group maythen be removed and any other modifications of the C-substituents effected. In particular R' mayfor 125 example represent an aliphatic hydrocarbon group, a hydroxyamine, such as 2 - hydroxyethylamine, or 3 - hydroxypropylamine, a diamine, such as ethylene diamine, Gran amino acid, sucKas glycine, (x- or P alanine,y-aminobutyric acid ortaurine. Hydroxy, GB 2 136 807 A 5 amineand carboxy (orsulpho) groups in N-substituents may of course be convertedto ester, amide and estergroups, respectively.

An alternative procedure involvesthe use of a 2 - aliphatic acyl 3 hydroxyfuran which may be reacted with a compound WNH2, the reaction, for example, of 2 - acetyl - 3 - hydroxyfuran with the sodium salt of glycine to form 1 - carboxymethyl - 3 - hydroxy - 2 methylpyrid -4 - one being described by Severin and Loidl in Z. Lebensm. Unters.-Forsch. 1976,161,119.

An example of the modification of C-substituents afterformation of the pyridone ring arises in the case where it is appropriate for substituents on carbon atoms of the ring to be protected whilst substitution is effected atthe nitrogen atom of the ring. Thus, for example, where a hydroxymethyl group is present on the ring as well as the 3 - hydroxy group, both hydroxy groups in a 3 - hydroxy - 4 - pyrone may be protected as described above. Moreover, N-acylated pyrid - 4 - ones may be prepared by direct acylation but it is preferred to protectthe 3 - hydroxy group, for example as an ether group such as in benzyloxy group, and to remove this protecting group afterthe N-acylation has been effected. A similarform of protection of the 3 - hydroxy group is suitable where a 6 hydroxyaikyl group is present which is being modified, for example to give a 6 - alkoxyalkyl or 6 haloalkyl group.

Itwill be appreciated from theforegoing that in many cases the final stage inthe preparation of the compound comprises deprotecting the hydroxy group of a pyridone having similar ring carbon atom substituents butwith the 3 hydroxy group in protected form and eitherthe same ring nitrogen atom substituent or one convertible therto and, where applicable, converting the N-substituentto that present in said compound and/or, optionally, converted the compound to a saitthereof containing a physiologically acceptable ion or ions. The compounds may be converted to salts formed between a physiologically

acceptable ion or ions and one or both of (a) the anion produced bythe loss of the hydroxy group proton and (b) an anionic or cationic group derived from an appropriate substi- tuent, such as one which is orwhich contains a carboxy, sulpho or amino g roup, by reaction of the compound with the appropriate base or acid according to standard procedures (amino substituted compounds of a zwitterion type containing a cation from the amino group and an anion from the 3 - hydroxy group may be prepared by crystallisation from aqueous media at a pH of about g).

In general, it is preferred thatthe compounds are isolated in substantially pureform, i.e. substantially freefrom by-products of manufacture.

ftwill be appreciated thatthese are notthe only routes available to these compounds and thatvarious alternatives may be used aswill be apparentto those skilled in the art, aswill bethe routesto the various intermediates required such as C-substituted 2,3and 3,4- dihydroxypyridines and 3 hydroxy - 4 pyrones.

Moreover, itwill be appreciated thatcertain of the compounds may be converted in vivo to other compoundswhich will be involved forthe metal 6 binding activity observed in vivo. This will be true, for example, of compounds containing ester groups which are likely to be converted to carboxy groups when the compounds are administered orally.

The compounds maybe formulated for use as pharmaceuticals forveterinary, for example in an avian or especially a mammalian context, 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 diluentwhich most usuallywill be employed for parenteral administration and therefore will be sterile and pyrogen free. However, itwill be appreciatedfrom 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, particularly in humans, to use composi- tions incorporating a solid carrier, forexample a conventional solid carrier material such as starch, lactose, dextrin or magnesium stearate. Compositions comprising a diluent of water andlor an organic solventwhich are non-sterile are therefore of less interest. Such solid compositions may conventiently be of a formed type, for example as tablets, capsules (including spansules), etc.

Otherforms 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 submultiple of a unit dose. Whilstthe 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 bodywill often be achieved using a daily dosage of aboutO.1 g to 5 g, particularly of aboutO.5 g to 2 g,veterinary doses being on a similarg/Kg body weight ratio. However, itwill be appreciated that it may be appropriate undercertain circumstancesto give daily dosages either below orabovethese levels. Where desired, more than one compound according to the present invention may be administered in the pharmaceutical composition or, indeed other active compounds may be included in the composition.

It has never before been appreciated that compounds such asthose described herein might be used in a pharmaceutical context, and with real advantage. We havefound thatthe3 - hydroxypyrid - 2 -and -4ones 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 cornplexes will partition into n-ocatanol indicating that they will permeate biological membranes, this property being confirmed in practice bytests of the ability of the"Fe labelled iron complexesto permeate erythrocytes.

Both the 3- hydroxypyrid - 2 - ones and the 3 - GB 2 136 807 A 6 hydroxypyrid -4- ones prossessa high affinityfor iron (111), as evidenced bylog K.,Olvalues {log K,,., is defined as being equal to log PFe(L)n + 21 - [pK.P + n log aL(H+) + m log aL(Ca++)jwhere log 0Fe(L)n isthe cumulative affinity constant of the ligand in question foriron (111), pKsp isthe negative logarithm of the solubility productforFe(OH)3and hasavalue of 39, n and m arethe numberof hydrogen andcalcium ions, respectively, which are boundtothe ligand,and aL(H+)and aL(Ca++) are the affinities ofthe ligandfor hydrogen ions and calcium ions, respectively}. In orderto solubilise iron (111) hydroxide, log K.,,, must be greaterthan 0 and in orderto remove iron from transferrin, log K.., should be in excess of 6.0. The log K,,., valuesfor3 - 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 about4.0, 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 compoundsto remove iron efficiently has been confirmed both by in vitrotests and also by in vivo tests in mice. It is particularly signif ica nt that these lattertests are successful whetherthe compound is given intraperitoneally or orally by stomach tube,the compounds generally either being stable under acidic conditions or being converted therebyto acid stable active compounds. Oral activity is not general- ly present among the othertypes of compound previously suggested for use as iron co-ordinating drugs and although certain EDTA analogues do show such activity, they possess drawbacksfor pharmaceutical use.

In addition tothe use described hereinbeforefor thetreatmentof general iron overload, the3 hydroxypyridones described herein are also of interestfor use in certain pathological conditions wherethere may be an excess of iron deposited at certain sites even though the patient does not exhibit a general iron overload, this being the case, for example, in certain arthritic and cancerous conditions. Indeed in some patients having such conditions, the patient may exhibit an overall anaemia and in a further UK patent application in our name of even date herewith the use is described of the metal-free compounds of the present invention in conjunction with an iron complex forthe treatment of such patients.Thus, a mixture of an iron complexof a3- hydroxypyridone as described herein together with oneofthe3- hydroxypyridones in metal-freeform will havethe effects of remedying the overall anaemia through the action of the iron complex whilstthe metal- free compound will actto remove iron from pathological to physiological sites.

Although the major use of the compounds is in the removal or iron, they are also of potential interestfor the removal of some other metals present in the body in deleterious amounts for example copper, pluto- nium and other related transuranic metals, and especially aluminium. The present invention thus includes the use of a 3 - hydroxypyrid - 2 - or -4- one or saItthereof as described hereinbeforefor use in medicineJor example forthe removal from the body of toxic amounts of metals, particularly iron. Moreov- 7 er, the invention alsaincludes a method forthe 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 - or4 - one or saitthereof as described hereinbefore to effect a reduction of the levels of this metal in the patient's body.

Uses of the compounds of the present invention for combination with metals otherthan iron may extend to the treatment of bodyfluids outside the body or even to quite other contextsthan thetreatment of patients. One particular area of some interest involvesthe treatment of patients on haemodialysis who may show a dangerous build up of aluminium in the body. Forthe treatment of such patients the compounds of the present invention may be attached to a support material and then contacted with the patient's blood to remove aluminium therefrom. The su pport material may conveniently be one of various types of polymer described in the artfor use in similar contexts, for example a carbohydrate material which may be of an agarose, dextran or other type, or a polystyrene or other material such as is used in ion- exchange resins.

Various approaches known in the art may be used for effecting attachment of the compounds to such support materials but one convenient approach isto use an acidic or basic group on the support material to provide an amide type linkagethrough reaction with the hydroxypyridone. Hydroxypyridones of particular interest in this context arethose containing acidic or basic substituents on a ring carbon or particularly on the a ring nitrogen atom. Of particular interest are hydroxypyridones which contain an N-substituent which is an aliphatic hydrocarbon 100 group substituted by an aliphatic amine or a sulpho or especially a carboxy group.

The present invention thus further comprises a 3 - hydroxy hydroxypyrid 2 - or - 4- one as defined hereinbefore linked to a support material through one of said substituents.

This invention is illustrated bythe following Examples.

EXAMPLES

Example 1 Thepreparation of 1 -acetyl-3- hydroxypyrid-2- one 2,3 Dihydroxypyridine (5.55 9) is refluxed with acetylbromide (10 m[) overnight. The reaction mixture is allowed to cool and water (10 m[) is added. The resulting suspension is extracted with methylene chloride and the methylene chloride solution is dried over Na2SO4and evaporated. The resultant residue is recrystallised from petroleum ether (80-1 OWC) to give 1 - acetyl - 3 - hydroxypyrid - 2 - one as white crystals (2.8 g), m.p. 140-141'C; v..,, (nujol) 1680,3120 cm-l; 5(d6DMSO), 2.2 (s, 3H), 6.1 (t, 1 H) 6.65 (m, 1 H), 7.2 (cl, 1 H). Example2 The preparation of 1 ethoxycarbonyimethyl - 3 60 hydroxypyrid-2-one 2,3 - Dihydroxypyridine (5 9) is suspended in ethyl bromoacetate (20 mi) and the mixture heated in a sealed tube for 24 hours at 1400C. Thetube is then cooled in solid C02 and opened. The contents are subjected to rotary evaporation at50OCto yield a GB 2 136 807 A 7 yellow solid. Recrystallisation of this solid f rom water yields 1 - ethoxycarbonyl methyl - 3 - hydroxypyrid - 2 - one as white crystals (5.4 g), m.p. 141-15loC; Vmax (nujol) 1645,1720 cm-l; 6(d6DMSO), 1.0 (t, 3H), 4.0 (q, 2H), 4.55 (s, 2H), 5.95 (t, 1 H), 6.6 (cl, 1 H), 7.05 (cl, 1 H). Example 3 The preparation of 1 - carboxymethyl - 3 hydroxypyrid-2-one 1 - Eth oxyca rbo nyl methyl - 3 - hydroxypyrid - 2 one (2 g) is dissolved in 15 m[ of 1: 2 vlv ethanollwater containing sufficient ammonium hydroxide to create a pH of 12.0. The solution is heated at WC for 30 minutes, cooled, and acidified to pH 3.0 by the addition of formic acid. The solution is rotary evaporated to remove most of the ethanol and then freeze dried for 20 hou rs to remove the water, formic acid and ammonium formate. The resulting solid is recrystallised from water to yield white crystals (1.4 g), m.p. 203-205'C; vm,x (nujol) 1540,1590,1640, 1695,3240 cm-l; 6(d6DMSO) 4.5 (s, 2H), 5.95 (t, 1 H), 6.6 (d, 1 H), 7.0 (d, 1 H). Example 4 The preparation of3- hydroxy- 1 methoxycarbonylmethylpyrid-2-one 1 - Carboxymethyl - 3 - hydroxypyrid - 2 - one (1 g), prepared as described under Example 3, is refluxed in methanolic hydrochloric acid for 2 hours. The solvent is removed by rotary evaporation and the residue recrystailised from waterto give 3 - hydroxy 1 - methoxyca rbonyl methyl pyrid - 2 - one in 60% yield, m.p. 141-142oC; vrn. x (nujol) 1560,1595,1645,1730, 3220 cm-l; 6(d 6 DIVISO) 3.55 (s, 3H), 4. 65 (s, 2H), 6.05 (t, 1 H), 6.7 (d, 1 H), 7.1 (cl, 1 H) 9.2 (s, 1 H). Example 5 The preparation of 1 - (N- ethylcarbamoyimethyl) -3 hydroxyprid- 2-one 3- Benzyloxy- 1 - carboxymethylpyrid-2 - one 1 - Eth oxyca rbonyl methyl - 3 - hydroxypyrid - 2 one (10 g), prepared as described under Example 2, is dissolved in methanollwater (9:1 vlv) (400 mi). To this solution is added benzyi chloride (3 molar excess) and NaOH until the pH is above 12. The mixture is then refluxed for six hoursto give a clear orange solution. The methanol is removed by rotary evaporation and the aqueous solution is extracted with dichloromethane to remove excess benzyl chloride. The aqueous phase is diluted slightly by adding extra water and then acidified to pH 2 using concentrated hydrochloric acid which results in the precipitation of a beige solid. The mixture is cooled and the precipitate filtered off and washed with diethyl ether. The crude product is recrystallised from ethanol to give 3 benzy[oxy - 1 - carboxymethylpyrid - 2 - one (5.4 g, 41 %), m. p. 176-177'C.

3 - Benzyfoxy - 1 - (succinimido - oxycarbonyimethyi) pyrid-2-one 3 Benzyloxy - 1 - ca rboxymethylpyrid - 2 - one (2 9) is dissolved in dimethylformamide (DMF) (25 mi) and tothis solution is added N hydroxysuccinimide (1 g).

The resultant solution is cooled and to it is added dicyclohexylearbodiimide (DCCI) (11.8 g) in DMF (5 m]). The mixture is allowed to stand overnightto give a brown super-natent and a white precipitate. The precipitate isfiltered off, washed with a little DM F and then evaporated to dryness under high vacuum. The 8 crude product is dissolved in a minimum volume of dichloromethane and diethyl ether is addedto this solution until it becomes cloudy.The solution isthen cooled andthe resultant precipitate isfiltered off and washed with a little diethyl etherto give 3 - benzyloxy - 1 - (succinimido - oxycarbonyl methyl) - pyrid - 2 - one (2.26 g, 82%), m.p. 187-188'C. 3 - Benzyloxy- 1 - (N - ethylcarbamoyimethyl) - pyrid 2-one 3 - Benzyloxy - 1 (succinimido - oxycarbony]methyl) - pyrid - 2 - one (1 g) is dissolved in dichloromethane (100 mi) and to this solution is added ethylamine in a 2 molar excess. The mixture is allowed to stand forfive minutes andthen the N - hydroxysuccinimide liberated bythe reaction is extracted with 1 M sodium bicarbonate (2 X 25 m[). The organic layer is dried overanhydrous sodium sulphate, filtered, and evaporated to give 3 - berixyloxy- 1 - (N ethyl carba moyl methyl) - pyrid - 2 - one as a greysolid in 50% yield, m.p. 171-172'C. 1 - (N- ethylcarbamoyimethyl) -3-hydroxypyrid-2one 3 Benzy[oxyl - (N - ethylcarbamoyl methyl) - pyrid 2 - one (1 g) is dissolved in 50:50 vlv aqueous ethanol.

Platinum/ca rbon catalyst (100 mg) is added and the solution is hydrogenated at WC and atmospheric pressure for eight hours. The mixture is filtered, the filtrate subjected to rota ry evaporation and the resulting residue recrystallised from ethanol to give 1 - (N - ethyl - ca rbamoyl methyl) - 3 - hydroxypyrid - 2 one in 70% yield, m.p. 214-215C, v,,,, (nujol) 1555, 1580,1645,1675,3260,3400 em-', 6(drDMSO) 1.0 (t, 3H), 3.05 (m, 2H), 4.5 (s, 2H), 6.05 (t, 1 H), 6.7 (d, 1 H), 7.05 (d, 1 H), 8.2 (s, 1 H).

Example 6 The preparation of3 - hydroxy- 1 (N - methylcarbamoyimethyl) pyrid-2- one 3 - Benzyloxy - 1 - (succinimido - oxycarbony]methyl) - pyrid - 2 one (1 g), prepared as described under Example 5, is reacted with methylamine under substantially similar conditions to those described under (E) forthe reaction of this compound with ethylamine to give 3 benzyioxy - 1 - (N - methylcarbamoyimethyi) - pyrid - 2 - one as a white solid in 84% yield, m.p. = 182-1830C.

This compound is hydrogenated as described under (E) forthe corresponding ethyl analogueto give 3 - hydroxy - (N - methyl - carbamoyl methyl) pyrid - 2 - one in 60% yield, m.p. = 204-205'C; vmax (nujol) 1560,1585,1645,1685,2950,3275 em-'. Mcl,13MS0) 2.40,2.46 (2 x s, 3H), 4.35 (s, 2H), 6.55 (d, 1 H), 6.95 (d, 1 H), 7.95 (s, 1 H). Example 7 Thepreparation of3-hydroxy- 1 - (Npropylcarba- moyimethyl) - pyrid - 2 one 3 - Benzyloxy - 1 - (succinimido - oxycarbonyImethyl) - pyrid - 2 one (1 g), prepared as described under Example 5, is reacted with n propylamine under substantially similar conditions to those de- scribed under Example 5 forthe reaction of this compound with ethylamine to give 3 - benzyioxy - 1 (N - propylcarbamoyimethyi) - pyrid - 2 - one as a beige solid in 60% yield, m.p. 156-157C. This compound is hydrogenated as described under Example 5forthe corresponding ethyl analogue to give 3 GB 2 136 807 A a hydroxy- 1 - (N - propylca rbamoyl methyl) - pyrid-2 one in 47% yield, m. p. 204-205'C; v,,x (nujol) 1560, 1580,1640,3050,3200 em-', 6(d6DMSO) 0.85 (t, 3H), 1.5 (m, 2H),10 (q, 31-1), 4.5 (s, 2H), 6.05 (t, 1 H), 6.7 (el, 1 H), 7.05 (d, 1 H), 8.1 (s, 1 H), 8.95 (s, 1 H). Example 8 The preparation of3 - hydroxy - 1 - [N - (2'methylethyl) carbamoyimethyll- pyrId-2 - one 3 - Benzyloxy - 1 - (succinimido oxycarbony- Iniethyl) -pyrid-2 -one (1 g), prepared asdescribed under Example 5, is reacted with isopropylamine under substantially similar conditions to those described under Example 5forthe reaction ofthis compound with ethylamineto give 3 - benzy[oxy- 1 [N-(2"-methylethyi)-carbamoyi-methyll-pyrid-2one. This compound is hydrogenated as described under Example 5- forthe corresponding ethyl analogueto give 3 - hydroxy- 1 -[N-(2'-methylethyi)carbamoyimethyll.pyrid -2- one as a silvery powder in 60% yield, m.p. 238-241'C; Vnax (nujol) 1570,1595, 1650,3270 em-'; 6(d6DMSO) 0.95 (cl, 6H), 3.7 (m, 1 H), 4.4 (s, 2H), 5.95 (t, 1 H), 6.53 (cl, 1 H), 6.98 (d, 1 H), 7.96 (d, 1 H), 8.85 (s, 1 H). Example 9 Thepreparation of 1 - (N -butylcarbamoylmethyl) -3hydroxy-pyrid-2-one 3 - Benzyloxy - 1 - (succinimido oxycarbonyImethyl) - pyrid - 2 - one (1 g), prepared as described under Example 5, is reacted with n - butylamine under substantially similar conditions to those described under Example 5forthe reaction of this compound with ethylamineto give 3 - benzyloxy - (N butylcarbamoyl methyl) - pyrid - 2 - one. This compound is hydrogenated as described under Example 5forthe corresponding ethyl analogueto give 1 - (N butylcarbamoyl - methyl) - 3- hydroxypyrid - 2 - one as colourless needles of m.p. 199200'C; vn,,x (nujol) 1565,1595,1650,1680,3100,3270 em-'; 6(d6DMSO), 0.8 (t, 3H), 1.3 (m, 4H), 3.0 (d, 2M, 4.43 (s, 2H), 5.95 (t, 1 H), 6.63 (cl, 1 H), 6.96 (d, 1 H), 8.0 (t, 1 H), 8.86 (s, 1 H). Example 10 Thepreparation of 1 -acetyl-3-hydroxy-2methylpyrid-4-one 3-Benzyloxy-2- methyl-4-py,rone 3 - Hydroxy- Z- methyl -4- pyrone (22.2 g) in methanol (225 mi) is added to aqueous sodium hydroxide (25 mi H20 containing 7.5 9 NaOH). Benzy] chloride (25.5 g) is added and the mixture is refluxed for 6 hours and isthen allowed to cool overnight. The bulk of the methanol is removed undervacuum and the residue istreated with water (50 mi). The mixture is extracted into, dichloromethane (3 X 25 ml). The, extracts are combined, washed with 5% w/v NaOH (2 x 25 m10hen water (2 x 25 mi) and dried over magnesium sulphate. Evaporation of the solvent gives crude3 - benzy[oxy 2 - methyl - 4 - pyrone- (35 g, 92%) which is purified by distillation in nitrogen under reduced pressureto yield a cQlourless oil (28 g) of b.p. 1480C10.2 mm. 3 Benzy[oxy-2- methylpyrid- 4 -one 3- Benzyloxy-2- methyl 4- pyrone (20 g), concentrated (s.g. 0.880) ammonia (200 mi) and ethanol (100 mO are mixed and kept at room temperaturefor3 days. The solvent and excess ammonia arethen removed by rotary evaporation to 1 1 9 yield an oil which on trituration with acetone gives 3 benzyloxy-2 - methylpyrid -4one as white crystals m.p. 162-16WC.

1 -Acetyl-3-benyloxy-2- methylpyrid-4- one 3- Benzyloxy-2- methylpyrid -4one (49) is 70 dissolved in dryacetone (100 mi).Acetyibromide (3 9) andtriethylamine (2.7 g) are added andthe resulting mixture is mechanically stirred overnight. The mix ture isthen filtered andthefiltrate is rotary evapo- rated to dryness. The residue is dissolved in methylene chloride and washed with dilute hydroch loric acid (pH 3.0), then twice with water, and is dried over Na2S04 and rotary evaporated to yield a solid residue. Recrystallisation of this residue from an ethyl acetate/hexane mixture gives 1 - acetyl - 3 - benzyloxy - 2 - methyl pyrid - 4 - one as an oil (3.1 g).

1 -Acetyl - 3 - hydroxy - 2 - methylpyrid- 4 - one 1 - Acetyl - 3 benzyloxy - 2 - methylpryid - 4 - one (2 9) is treated with 45% w/v HBr acetic acid (10 m 1) for 1 hour at 1 OWC. The solution is rotary evaporated to dryness at7WC and triturated with an ethyl acetate/ methanol (20.1 vlv) mixture. On standing overnight at 4'C pale brown crystals are deposited (1.2 g) and these are recrystaffised from an etbyl acetatelmetha no] mixtureto yield 1 - acetyl - 3 - hydroxy - 2 - methylpyrid -4- one aswhite crystals, m.p. 152 160OC; vm,x (nujol) 1630,1680 cm-l; 6(d6DIVISO), 2.2 (s, 3H), 2.5 (s, 3H), 7.25 (d, 1 H), 8.15 (d, 1 H).

Example 11

The preparation of3 - hydroxy- 1 - (2'- hydroxyethyP 3 - Benzyloxy - 1 (2'- hydroxyethyi) - 2 - methylpyrid - 4-one 3 - Benzyloxy - 2 - methyl 4 - pyrone (4.8 g), prepared as described under Example 10, and 2 hydroxyethylamine (1.22 g) are dissolved in water (220 mi) and ethanol (100 m]) containing sodium hydroxide (2 g) is added. The mixture is stirred at room temperaturefor6 days and isthen 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 mi). The chloroform extracts are combined, dried over magnesium sulphate, and evaporated to yield 3 - benzyloxy - 1 - (2'hydroxyethyl) - 2 - methyl - pyrid - 4 - one, as a white solid (3.4 g), m.p. 198-199'C.

3-14ydroxy- 1 - (2'- hydroxyethyl) - 2 - methylpyrid-4 - one 3 - Benzyloxy - 1 - (2'- hydroxyethyl) - 2 - methylpyrid -4- one (2 9) is added to concentrated hydrobromic acid (10 mi) and the mixture is heated on a steam bath for 30 minutes. The resultant product is then recrystallised from waterto yield 3 - hydroxy - 1(2'- hydroxyethyl) - 2 - methylpyrid -4 - one as white crystals (0.8 g), m.p. 164-165OC; vm,, (nujol) 1630, 3150,3350 cm-l; 6(dbDMSO), 2.5 (s, 3H), 3.7 (t, 21-1), 4.35 (t, 2H), 7.25 (3, 1 H), 8.15 (d, 1 H).

Example 12

Thepreparation of3- hydroxy- I- (Y- hydroxy propyi) - 2 -methyl -pyrid-4 -one 3- Benzyloxy-2- methyl -4- pyrone, prepared as described under Example 10, is reacted with 3 - hydroxypropylamine under substantially similar con ditions to those described under Example 9 for reaction with 2 - hydroxyethylamine to give 3 - 130 GB 2 136 807 A 9 benzyloxy - 1 - (Xhydroxypropyl) - 2 - methylpyrid - 4 - one. This is deprotected using the procedure described under (1) to give 3 - hydroxy - 1 - (Xhydroxypropyl) 2 - methylpyrid - 4 - one as white crystals, m.p. 111-1 1WC; vm,, (nujol) 1630,3150,3350 cm-1, 6(d6DIVISO) 1.8 (m, 2H), 2.4 (s, 3H), 3.35 (t, 2H), 4.33 (t, 2H), 7.3 (cl, 1 H), 8.2 (cl, 1 H). Example 13 Thepreparation of3-hydroxy- 1- (4'-gydroxybutyl) -2-methylpyrid-4-one 3 - Benzyloxy - 2 methyl - 4- pyrone, prepared as described under Example 10, is reacted with 1 - amino - 4 - hydroxybutane under substantially similar conditions to those described under Example 11 for reaction with 2 hydroxyethyiamine to give 3 benzyloxy - 1 - (4- hydroxybutyl) - 2 methylpyrid - 4 one. This is deprotected using the procedure described under (1) to give 3 - hydroxy - 1 - (4'hydroxybutyl) - 2 - methylpyrid - 4 - one as white crystals, m.p. 126-128'C; Vnax (nujol) 1630,3350 cm-i; 6(d6DIVISO) 1.5 (m, 4H), 2.45 (s, 3H), 3.35 (t, 2H), 4.30 (t, 2 H), 7.25 (cl, 1 H), 8.2 (cl, 1 H). Example 14 The preparation of3 - hydroxy- 1- (Yhydroxypen90 tyl) - 2- methylpyrid- 4 - one 3 - Benzyloxy - 2 - methyl - 4 - pyrone, prepared as described under Example 10, is reacted with 1 amino - 5 - hydroxypentane under substantially similar conditionsto those described under Example 11 for reaction with 2 - hydroxyethylamine to give 3 benzy[oxy - 1 - (W- hydroxypentyl) - 2 - methyl pyrid - 4 - one. This is deprotected using the procedure described under (1) to give 3 - hydroxy 1 - (Whydroxypentyl) - 2 - methylpyrid - 4 - one as white crystals, m.p. 136-138OC; vn,,x (nujol) 1625,3350 cm-i: 6(d6DIVISO) 1.3 (m, 61-1), 2.40 (s, 31-1), 3.25 (t, 2H), 4.20 (t, 21-1), 7.20 (d, 1 H), 8. 15 (d, 1 H). Example 15 Thepreparation of I- (5-acetoxypentyl) -3-hydroxy 105 -2-methylpyrid-4-one 3 - Hydroxy - 1 - (W- hydroxypentyl) - 2 methylpyrid - 4 - one, prepared as described under Example 14,(2 g) is dissolved in glacial acetic acid containing about 1 % w/v of hydrogen bromide and the solution is refluxed for 2 hours. The resultant mixture is subjected to rotary evaporation and the residue crystallised from aqueous ethanolto give 1 (5'-acetoxypentyi)-3-hydroxy-2-methylpyrid-4one in 65% yield, m.p. 132-1330C, Vmax (nujol) 1520, 1540,1580,1635,1735 cm-1 and 6(d6DIVISO), 1.6 (m, 6H), 2.1 (s, 3H), 2.4 (s, 3H), 4.05 (m, 4H), 6.4 (cl, 1 H), 7.3 (cl, 1 H). Example 16 Thepreparation of 1 - carboxymethyl - 3 - hydroxy- 2 120 methylpyrid-4-one 3 - Benzyloxy - 2 - methyl - 4 - pyrone, prepared as described under Example 10, is reacted with glycine under substantially similar conditions to those described under Example 11 for reaction with 2 - hydroxyethylamineto give 3 - benzyloxy - 1 carboxymethyl - 3 - hydroxy 2 - methylpyrid - 4- one, which is deprotected using the procedure described under Example 11 to give 1 - carboxymethyl -3 hyd;oxy- 2 methylpyrid -4- one aswhite crystals, m.p. >230'C; vmax (nujol) 1625,1645 cm-1 GB 2 136 807 A 10 6(d,DMSO) 1.9 (s, 3H),4.3 (s, 2H), 5.9 (d, 1 H),735 (d, 1 H).

Example 17

Thepreparation of 1- (2'-carboxyethyl) -3hydroxy 2-methylpyrid-4 -one hydrochloride 3-Benzyloxy- 1-(2'-carboxyethyt)-2-methylpyrid- 4-one 3Benzyloxy-2- methyl -4- pyrone, prepared as described under Example 10, (20 g) and beta - alanine (9 g) are dissolved in 3:2 vlvwaterlethanol (500mi) containing NaOH (10 9)to provide a solution with a pH of at least 13. The solution is refluxed for 15 minutes whereupon itturnsfrom a light orangeto an intense red colour. The solution is acidified to pH 7.0 and the ethanol is removed by rotary evaporation.

The resulting aqueous solution is washed twice with ethylacetate (100 m]). This solution is then subjected to rotary evaporation until the volume is reduced to mi and this reduced volume of solution is 2a acidified to pH 3.0 to give a white precipitate, which is 85 filtered off to give 3 benzyioxy - 1 - (2'- carboxyethyl) - 2 - methylpyrid - 4 - one in 70% yield, m.p. 156-157'C.

1- (2'- Carboxyethyl) -3- hydroxy-2 - methylpyrid-4 - one hydrochloride 3 - Benzyloxy - 1 - (2'- carboxyethyl) - 2 - methyl pyrid - 4 - one is subjected to hydrogenation in aqueous ethanol (1: 1 vlv) in the presence of platinum/ carbon catalyst (100 mg per gram of pyridone) for 8 hours at 2WC and atmospheric pressure. Filtration, followed by rotary evaporation of thefiltrate, yields a 95 white solid which on recrystallisation from acetone and diethylether gives 1 - (2'- carboxyethyl) - 3 - hydroxy - 2 methylpyrid - 4 - one hydrochloride in 55% yield, m.p. <40'C: v,,,,x (nujol) 1590,1620,1720 cm-l: 6(d6DIVISO) 2.5 (s, 31-1), 2.8 (t, 21-1), 4.45 (t, 2H), 7.35 (d, 1 H), 8.2 (d, 1 H).

Example 18

Thepreparation of 1-(2'-ethoxycarbonylethyl)-3 hydroxy- 2 - methylpyrid- 4 - one 1 - W- Carboxyethyl) - 3 - hydroxy - 2 - methylpyrid - 4- one hydrochloride (2 g), prepared as described under Example 17, is dissolved in ethanol saturated with hydrogen chloride and the solution is refluxed for 3 hours. Rotary evaporation of the solution yields a white solid which on recrystallisation from ethanol gives 1 - (2'ethoxycarbonylethVI) - 3 - hydroxy - 2 - methylpyrid - 2 - one hydrochloride in 75% yield, m.p.

127-130OC; vm,x (nujol) 1530,1620,1720 cm-l; 5(d6DMSO) 1. 1 (t, 31-1), 2.5 (s, 31-1), 2.9 (t, 2H), 4.0 (q, 21-1), 4.5 (t, 2H), 7.4 (d, 1 H), 8.2 (d, 1 H).

Example 19

Thepreparation of3 - hydroxy- 1 - (2'-methoxycar bonylethyl) -2methylpyrid- 4 - one 1 - (2'- carboxyethyl) - 3 - hydroxy - 2 -methylpyrid - 4- one hydrochloride (2 g) prepared as described under Example 17, is dissolved in methanol saturated with hydrogen chloride and the solution is refluxed for3 hours. Rotary evaporation of the solution yields a white solid which on recrystallisation from metha nol gives3 hydroxy - 1 - (2- m ethoxyca rbonyl ethyl) - 125 2- methylpyrid -4 - one hydrochloride in 80% yield m.p. 140-14M, Vmax 1545,1595,1645,1730,1750 cm-1, 6(d6DMSO), 2.45 (s, 31-1), 2.85 (t, 2H), 3.5 (s, 31-1), 4.45 (t, 2H), 7.30 (d, 1 H), 8.15 (d, 1 H).

Example 20

The preparation o f3 - h ydroxy - 1 - [2' - (N - p rop ylca rbam o yO - e th y/1 - 2 - m eth ylpyrid - 4 - on e 3 - Benzyloxy - 1 - [2'(succinimido - oxycarbon yl) - ethyl] - 2 - meth ylpyrid - 4 - one 3 - Be nzy[oxy - 1 - (2' - ca rb oxyethy 1) - 2 - methyl pyrid - 2 - one, prepared as described under Example 17,(2.2 9) is dissolved in DM F (25 mi) and to the solution is added N - hydroxysuccinimide (1 g).

This solution is cooled and to it is added dicyclohexyl- carbodiimide (1.8 g) in DIVIF (5 mi). The mixture is allowed to stand overnight giving a dark coloured solution and white precipitate. The precipitate is removed byfiltration and washed with DIVIF (3 mi). The solution is rotary evaporated to dryness and triturated with diethyletherto give 3 benzy[oxy - 1 [2'- (succinimido - oxycarbonyl) - ethyl] - 2 methylpyrid 4 - one hydrochloride as a white solid in 55% yield, m.p. 45-47'C. 3 Benzyloxy - 1 - [2'- (N - propylcarbamoyl) - ethyl] 2 -methylpyrid-4-one 3 - Benzy] oxy - 1 - [2' - (succi nimido - oxycarbonyl) ethyl] - 2 methylpyrid - 2 - one (1 g) is dissolved in chloroform (100 m]) and to this solution is added ethylamine in a 2 molar excess. The mixture is al lowed to stand for 15 minutes and the N hydroxysuccinimide liberated bythe reaction isthen extracted with 1 M Nal-IC03solution (2 x 25 mi). The organic layer is dried over Na2S04, filtered and evaporatedto drynessto give 3 - benzyloxy- 1 - [2'- (N -propylcarbamoyl) - ethyl] -2 methylpyrid -4- one as a white solid in 40% yield, m.p. 146-147'C. 3 Hydroxy- 1 - [2'- (N - propylcarboamoyl) - ethyl] - 2 -methylpyrid-4one 3 - Benzyloxy - 1 - [2'- N - propylcarbamoyl) - ethyl] - 2 - methylpyrid -4one (5 g) is dissolved in 50:50vlv aqueous ethanol. Platinum/carbon catalyst (100 mg) is added and the solution is hydrogenated at 200C and atmospheric pressure for 8 hours. The mixture is filtered, thefiltrate subjected to rotary evaporation and the resulting residue recrystallised from ethanol to give 3 - hydroxy- 1 - [2'- (N - propylcarbamoyl) ethyl] pyride4- one, m.p. 113-114'C,V, ax(nujol) 1505,1550,1570,1630,1705,3080,3200 cm-l; Md,DIVISO) 0.6 (t, 31- 1), 1.2 (sextuplet, 21-1), 2.5 (s, 31-1), 2.6,2.8 (overlapping t and q, 41-1), 4.5 (t, 2H), 7.2 (d, 1 H), 8.1 (overlapping d and t, 2H). Example21 The preparation of 1 - (2'- aminoethyl) - 3 - hydroxy -2 - methylpyrid - 4 one hydrochloride 3 - Benzyloxy - 2 - methyl - 4 - pyrone prepared as described under Example 4,(4 g), and ethylene diamine (1.5 g) are heated together under refl ux in water (50 mi) and ethanol (17 m[) for 1 hour. The solvent is removed by rotary evaporation and the solid residue is heated at 800C with concentrated hydrochloric acid (50 m 1) for 30 minutes. Excess acid is removed by rotary evaporation at 800C and the residue is slurried with acetone, yielding a pale brown solid (2. 2 g). Recrystallisation from ethanol, containing a trace of hydrochloric acid yields 1 - (2'aminoethyl) - 3 - hydroxy - 2 - methylpyrid - 4- one hydrochloride aswhite crystals, m.p. 2800C (with decomposition); vna,. (nujol) 1620,3150 cm-l; 5(1320) 2.5 (s, 3H), 3.5 (t, 2H), 4.5 (t, 2H), 7. 1 (d, 1 H), 8.1 (d, 1 H).

Example 22

W 1 4 11 Thepreparation of3-hydroxy-6-hydroxymethyl- 1 methylpyrid-4-one 3 - Benzyloxy- 6 - benzyloxymethylpyrid - 4 -one Kojic acid (3- hydroxy-6hydroxymethylpyrid -4- one) (11.5 g) in methanol (90 mi) is added to an aqueous solution of sodium hydroxide (3 g in 10 mi). Benzylchloride (10.2 g) is added and the mixture is stirred and refluxed for 6 hours. On cooling, crystals are obtained which are recrystallisated from metha- nol to give 3 - benzyloxy - 6 - benzyloxymethylpyrid - 4 -one aswhite crystals (10 g), m.p. 103-131'C; 6(CDC13)4.25(s,2H),4.99(s,2H),6.25(s,1H), 7.3 (s, 5H), 8.1 (s, 1 H). 3-Hydroxy-6hydroxymethyl- 1 -methylpyrid-4- one 3 Benzyloxy-6 - benzyloxypyrid -4- one (5.0 g) and methylamine hydrochloride (1.56 g) are dissolved in aqueous ethanol (300 m[ of 2:1 H20-CM,0H byvolume containing 2 g of sodium hydroxide). The reaction mixture isstirred at room temperatureforiS days and then acidified with concentrated HCI to pH 2.0. Theyellow mixture is evaporated to dryness and the resulting solid residue is heated under refluxwith concentrated HCl (50 ml) for30 minutes. The product is rotary evaporated to drynessyielding a brown residuewhich ontrituration with acetone forms crystals. Recrystallisation of thesefrom ethanol gives 3- hydroxymethyl - 1 methylpyrid -4- onein 15% yield, m.p. 185-18WC; 5(d6DMSO), 3.9 (s, 3H), 4.6 (s, 2H), 7.4 (s, 1 H), 8.3 (s, 1 H). Example23 The preparation of 1 ethyl - 3 - hydroxy - 6 - hydroxy methylpyrid-4-one 3 - Benzy[oxy - 6benzyloxypyrid -4- one, prepared as described under Example 22, is reacted with ethyia'i-nine hydrochloride under substantially similar conditionsto those described under Example 22 for reaction with methylamine hydrochloride to give 1 - ethyl - 3 - hydroxy - 6 - hydroxymethylpyrid - 4 - one as white crystals, m.p. 143-145'C; 5(d6DMS0) 1.1 (t, 3H), 4.0 (q, 2H), 4.3 (s, 2H), 7.2 (s, 1 H), 8.1 (s, 1 H). Example 24 The preparation of 1- (2'- aminoethyl) -3 - hydroxypyrid - 4 - one 3 - Benzyloxy - 4 pyrone is prepa red from 3 hyd roxy - 4 - pyrone as described by Spenser eta], Canadian Journal of Chemistry, 1962,40,1377 and has m.p. 82-85'C. A mixture of this compound (19) and ethylenediamine (0.37 g) in water (12 mi) is heated under refluxfor 1 hour. The reaction mixture is evaporated to dryness and the residue is treated with concentrated hydrochloric acid (12 m]) and the mixture is heated on a stem bath for 30 minutes. The excess acid and water are evaporated under reduced pressureto give a solid brown residue. This is slurried in acetone,then filtered off and reerystallised from ethanol containing a little concentrated hydrochloric acid to give 1 - (2- aminoethyl) - 3 - hydroxypyrid - 4 - one in 34% yield as colourless needles, m.p. 26426WC, vrnax (nujol) 1505, 1540,1600,1635 cm-1, 6(d6DMSO), 2.85 (t, 2H), 4.35 (t, 21-1), 7.1 (d, 1 H), 8.1 (m, 2H). Example25 Partition data on 3 - hydroxypyrid- 2 - and- 4 -ones GB 2 136 807 A 11 and their iron complexes The partition coefficient K,.r,, being the ratio (concentration of compound in n - octanolAconcentration of compound in aqueous phase) on partition between n - octanol and aqueous tris hydrochloride (20 mM, pH 7.4), is measured by spectrophotometry at 20'Cfor various of the compounds of the previous Examples and fortheir 3:1 iron (111) complexes (at 10-4M) by spectro photometry. The solutions of the complexes are either produced by dissolving the pre-formed complex in the aqueoustris hydrochloride or are prepared in situ in the buffer by the admixture of a 3:1 molar ratio of the pyridone and ferric chloride, the pH thereafter being readjusted to 7.4 if necessary (the same product will be obtained in the buffer solution irrespective of whetherthe pyridone used is in a saitform or not). Acid washed glassware is used throughout and, following mixing of 5 mi of the 10-4 M aqueous solution with 5 mi 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. Forthe free hydroxypyridones,the range 220-340 rim is used for concentration determinations whilstforthe iron complexes, the range 340-640 rim is used.

Values typical of those obtained are shown in Table 1.

12 GB 2 136 807 A 12 Table 1 Partition coefficients Partition coefficient K part Compound Free compound Iron complex [Fe ill- (compound) 31 1-acetyl-3-h%dr?xypyr;g-2-,one 0.55 0.24 1-ethoxycai)onylmethyl-3- 0.5 0.19 hydroxypyrid-2-one 3-hydroxy-1-methoxycarbonyl- 0.2 0.02 methylpyrid-2-one 1-(N-ethylcarbamoylmethyl- 0.12 0.01 3-hydroxypyrid-2-one 3-hydroxy-l-(N-methylcarbamoyl- 0.06 0.005 methyl)-pyrid-2-one 3-hydroxy-l-(N-propylcarbamoyl- 0.35 0.14 methyl)-pyrid-2-one 3-hydroxy-l-[N-(2'-methylethyl)- 0.34 0.15 carbamoylmethyll-pyfid-2-one 1-(N-butylcarbamoylmethyl)- 1.89 5.12 3-hydioxypyrid-2-one 3-hydroxy-l-(2'-hydroxyethyl)- < 0.002 < 0.002 2-methylpyrid-4-one 3-hydroxy-l-(3'-hydroxypropyl)- 0.10 0.06 2-methylpyrid-4-one Partition coefficient K part Compound Free compound Iron complex [Fe ill -(compound) 31 3-hyroxy-l-(4-hydroxybutyl)- 0.14 0.03 2-methylpyri&-4-one 3-hydroxy-l-(5'-hydroxypentyl- 0.28 0.04 2-methylpyrid-4-one 1-(5'-acetoxypentyl-3-hydroxy- 0.45 0.03 2--methylpyrid-4-one 1-carboxymethyl-3-hydroxy-2- < 0.002 < 0.002 methylpyrid-4-one 1-(2'-carboxyethyl)-3-bydroxy- 0.45 0.03 2 thylpyrid-4-one 1-(2-aminoethyl-3-hydroxy-2- < 0.002 < 0.002 methylpyrid-4-one 3-hydroxy-6-hydroxymethyl-l- 0.03 0.01 methylpyrid-4-one 1-ethyl-3-hydroxy-6-hydroxymethyl 0.06 0.07 pyrid-4-one Example26 In vitro tests of iron binding capacity The 3 - hydroxypyridones used in this Example were prepared as described in various of the previous Examples. (1) Mobilisation of iron from ferritin Horse spleen ferritin (Sigma) was used without further purification and its iron contentwas estimated spectrophotometrically at 420 nm. The ferritin solu- tion in phosphate buffered saline (Duibecco - OX011), 10-6M, pH 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 (111) corn- plex in the dialysis solution was recorded after 6 and 24 hours. For comparative purposes, the procedure was repeated using a blank control.

1 1 j The results are shown in Table 2 where the percentage offerritin-bound iron removed bythe compound under testis shown. For comparative purposes, results reported in the literature for similar tests with 1 X 10-3 M desferrioxamine (Crichton etal, J. Inorganic Biochem., 1980,13,305) and with 6 x 10-3M LICAMS (Tufano etal, Biochem. Biophys. Acta, 1981, 668,420) are also given in the Table. Itwill be seen thatthe pyridone compounds are able to remove iron effectively from ferritin in contrastwith desferrioxamine and LICAMS (although the latterwill remove iron in the presence of ascorbic acid such a mixture isvery difficuitto manage clinically). These results shown in Table 2 have been confirmed by separating apoferritin (in admixture with ferritin) and the particular hydroxypyridone iron (111) complex from the reaction product in each case by chroma- 13 tography on Sephadex G10.

(2) Mobilisation of iron from transferrin Human transferrin (Sigma) was loaded with iron (111) bythe method of Bates and Schiaback, J. Biol.

Chem. (1973) 248,3228. 9ron (111) transferrin (1 0-51V1) 20 was incubated with a 4 x 10-3M solution in tris HCI (0.1 M, pH7.4) of one of various pyridones as indicated in Table 3 for periods of 4 hours and 18 hours. The solution was then dialysed against phosphate buffered saline for 24 hours. The 5917e remaning in the dialysis tube was then recorded. For comparative purposes, this procedure was repeated with desforrioxamine using incubation for both 4 hours and 18 hours and with EDTA using incubation for4hoursonly.

The results are shown in Table 3 in terms of the Table 3

GB 2 136 807 A 13 Table 2

Removal ofiron from ferritin Percentage of iron removed Compound 6 hours 24 hours Control 0 0 1-acetyl-3-hydroxypyrid-2-one is 24 3-hydroxy-l-(21-hydroxyathyl)- 31 49 2-methylpyrid-4-one 1-carboxymethyl-3-hydroxy- < 5 < 5 2-methylpyrid-4-one 3-hydroxy-l-(2'-methoxycarbanylethyl)- 22 45 2-methylpyrid-4-one 1-(2'-aminoethyl)-3-hydroxy- 28 61 2-methylpyrid-4-one 3-hydroxy-6-hydroxymethyl- 25 49 1-methylpyrid-4-one Desferrioxamine (lmM) 1.5 - LICAMS (6mM) 0 LICAMS (6mM+12 mM ascorbic acid) 7 percentage of transferri n bou n cl i ro n removed by the corn pou nd u nder test. It wi 11 be seen that th e pyrid - 4 o ne cc m po u nds a re very effective at iron rem ova 1, as corn pa red with desferrioxa m i ne or EDTA, after on ly 4 hou rs. Althou g h the efficiency at i ron rem ova 1 of the pyrid - 2 - one corn pou nds is on ly at a sim i [a r level to that of desferrioxa m i ne a nd E DTA after 4 hou rs, it increases m a rked ly after 18 hou rs whereas th e level fo r desferrioxam ine at 18 hou rs is su bsta ntia 1 ly si m i la r to that at 4 hou rs.

Th e resu Its sh own i n Ta ble 3 h ave been co nfi rmed by sepa rati ng a potra nsferri n (i n adm ixtu re with tra nsfe rri n) a nd the pa rticu la r hydroxypyridione i ro n complex from the reaction product in each case by chromatography on Sephadex G1 0.

Removal ofiron from transferrin Percentage of iron removed Compound 6 hours 24 hours 1-acetyl,C3-hydroxypyrid-2-one is 30 1-carboxymethyl-3-hydroxypyrid-2-one 35 3-hydroxy-lmethoxycarbonylmethyl- 18 pyrid-2-one 3-hydroxy-l-[N-(2'-methylethyl)- 41 - carbamoylmethyll-pyrid-2-one 3-hydroxy-l-(2'-hydroxyethyl)- 94 96 2-methylpyrid-4-one 1-carboxymethyl-3-hydroxy- 82 88 2-methylpyrid-4-one 1-(21-aminoethyl)-3-hydroxy 97 98 2-methylpyrid-4-one 3-hydroxy-6-hydroxymethyl- 93 97 1-methlpyrid-4-one Desferrioxamine 17 22 EDTA 27 67 14 GB 2 136 807 A 14 Example27

In vivo tests ofiron binding capacity Th e 3 - hyd roxypyridones used i n th is Exa m p] e were prepared as described in various of the previous Examples.

Mice were injected intraperitoneally with iron 25 dextran (2 mg) at weekly intervals over a fourweek period. Two weeks afterthe final injection, the mice were injected via the tail vein with 59Fe lactoferrin (human lactoferrin, 1 mg per injection 2 1.0). The micewere then caged individually. After a ten day period, 1 - acetyl - 3 - hydroxypyrid - 2 one was administered to groups of 8 mice at 10 mg per mouse either intraperitoneally or intragastrical ly (in each case 3 of the mice received only one dose whilst 5 received 2 doses at a 24 hour interval. The excretion 35 of iron was recorded at either 12 or 24 hourly intervals over a th ree day period before and a two day period after administration of the compou nd. For compara- tive purposes, the procedure was repeated with a blank control and with desferrioxamine, also at 10 mg per mouse (the intraperitoneal ly treated mice receving one dose of desferrioxamine and the intragastrically treated mice two doses at a 24 hour interval.

The results are show in Table 4, being given on the basis of the control representing 100% excretion, and illustratethe particular advantage of the pyridones as compared with desferrioxaminefor oral administration the levels of iron excreted on intragastric administration being higherfor all the pyridones tested than for desferrioxamine. Itshould be mentioned thatthe large standard deviation (SID) values are somewhat misleading as uniforrnly positive results can yield high SDs which might be taken to suggestthatthe result are not significantly different from zero. However, this is notthe case here, the large SD values being a consequence of the large range among the positive responses.

Table 4

Excretion ofiron in vivo Intraperitoneal Intragastric Administration Administration Compound 7-b r Excretion Number Excretion f of 59Fe + SD of of 59 Fe SD MicT percent Mice percent Control 12 loo 10 1-acetyl-3-hydroxy- 8 136 33 8 137 45 pyrid-2-one 1-ethoxycarbonylmethyl12 90 11 12 92 13 3-hydroxypyrid-2-one 3-hydroxy-l-(2'-hydroxy- 11 170 49 11 17 -50 ethyl)-2-methylpyrid 4-one 3-hydroxy-l-(5'-hydroxy 9 149 59 9 111 22 pentyl)-2-methyl pyrid-4-one 1-carboxymethyl-3-hydroxy- 6 96 11 5 99 1o 2-methylpyrid-4-one 1-(2'-aminoethyl)- 12 115 16 12 100 14 3-hydroxy-2-methylpyrid 4-one Desferriaxamine 5 414 1 100 4 88 17

Claims

1. Acompound being a 3 - hydroxypyrid - 2 -one or 3 - hydroxypyrid - 4one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic acyl group, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substi- tuted by one or, except in the case of ionisable groups, morethan one substituent selected from aliphatic acy], alkoxy, aliphatic amine, aliphatic amide, carboxy, aliphatic ester, halogen, hydroxy and sulpho groups and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms are replaced by one of said substituents, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substituted by an alkoxy, aliphatic ester, halogen or hydroxy group, but excluding compounds in which said replacement of hydrogen 1 i 1 atoms in the compound is effected only by aliphatic hydrocarbon groups, ora saitthereof containing a physiologically acceptable ion or ions, for use in medicine.

2. Acompound being a 3- hydroxypyrid-2- one or3 - hydroxypyrid -4- one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphaticacyl group, byan aliphatic hydrocarbon group, or byan aliphatic hydrocarbon group substi- tuted by one or, except in the case of ionisable groups, more than one substituent selected from aliphatic acyl, alkoxy, aliphatic amine, aliphatic amide, carboxy, aliphatic ester, halogen, hydroxy and sulpho groups and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms are replaced by one of said substituents, by an aliphatic hydrocarbon group, or by an aliphatic i hydrocarbon group substituted by an alkoxy, alipha ticester, halogen or hydroxy group, but excluding compounds in which said replacement of hydrogen atoms in the compound is effected only by aliphatic hydrocarbon group and also the specific compounds 3 - hydroxy - 6 - hydroxymethyl - 1 - methylpyrid - 4 - one, 1 - W- aminoethyl) - 3 - hydroxypyrid - 4 - one, 3 - hydroxy - 1 (T- hydroxyethyl) - 2 - methylpyrid - 4 - one, 1 carboxymethyl - 3 - hydroxy - 2 - methylpyrid - 4 - one and 1 - ethoxy carbonyl methyl - 3 - hydroxy - 2 - methylpyrid - 4 - one, or a salt thereof with a physiologically acceptable cation.

3. A compound according to Claim 1 or 2, in which the hydrogen atoms attached to ring carbon atoms either are not replaced or are replaced by an aliphatic 80 hydrocarbon group.

4. A compound according to Claim 3, being a 3 - hydroxypyrid - 2 -one, 3 hydroxypyrid -4- one, 2 - alkyl -3- hydroxypyrid -4- one, 6-alkyl -3 hydroxypyrid -4 -one, or 2,6 -dial kyl 3 - hydroxypyrid nitrogen is thereof replaced by an aliphatic acyl group or by an aliphatic hydrocarbon group substituted by one or, except in the case of ionisable groups, more than one substituent selected from aliphatic acyl, alkoxy, aliphatic amine, aliphatic amide, carboxy, aliphatic ester, halogen, hydroxy and sulpho groups.

5. A compound according to Claim 3 or 4, in which the or each aliphatic hydrocarbon group or alkyl group replacing a hydrogen atom attached to a ring carbon atom is an acylic alkyl group of 1 to 4 carbon atoms.

6. A compound according to Claim 5, in which the or each acyclic alkyl group is methyl.

7. A compound accroding to any of the preceding claims, in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic acyl group -COR1, an aliphatic hydrocarbon group -(CH2)ncarrying a terminal substituent group -COR2, -COXR2, -SO2XR2, _XCOR2 or -XS02R,, or an aliphatic hydrocarbon group -(CH2)m- carrying a terminal substituent group -XH, wherein R, is an aliphatic hydrocarbon group, R2 is hydrogen on an aliphatic hydrocarbon group, X is an oxy or imino group, n is an integerfrom 1 to 6, and m is an integer from 2 to 6.

8. A compound according to Claim 7, in which R, and R2are each an acylicalkyl group of 1 to 4carbon atoms, n isan integerfrom 1 to4and m isan integer from 3 to 6.

9. A compound according to Claim 1 or 2, being a 3-hydroxypyrid -2-one N -substituted bya group -(CH2)nCOXR,ora3-hydroxy-2-methylpyrid-4- one N-substitdted bya group -(CH2)nCOXR1 or -(CH2)mM wherein R, is an acyclic alkyl group of 1 to 120 4 carbon atoms, X is an oxy g roup, n is an integer from 1 to 4 and m is an integerfrom 3 to 6.

10. A compound according to Claim 1, in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic hydrocarbon group and the hydrogen atom attached to the ring carbon atom at the 6-position is replaced by an alkoxymethyl, halomethyl or hydroxymethyl group.

11. A compound according to Claim 10, in which the aliphatic hydrocarbon group is an acyclic alkyl GB 2 136 807 A 15 group of 1 to 4carbon atoms.

12. A pharmaceutical composition comprising a 3 - hydroxypyrid -2 -one or3- hydroxypyrid -4- one in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic acyl group, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substituted by one or, except in the case of ionisable groups, morethan one substituent selected from aliphatic acyi, alkoxy, aliphatic amine, aliphatic amide, carboxy, aliphatic ester, halogen, hydroxy and sulpho groups and, optionally, in which one or more of the hydrogen atoms attached to ring carbon atoms are replaced by one of said substituents, by an aliphatic hydrocarbon group, or by an aliphatic hydrocarbon group substituted by an alkoxy, aliphatic ester, halogen or hydroxy group, but excluding compounds in which said replacement of hydrogen atoms in the compound is effected only by aliphatic hydrocarbon groups, or a saitthereof containing a physiologically acceptable ion or ions, together with a physiologically acceptable diluent or carrier.

13. A pharmaceutical composition according to Claim 12, in which the hydrogen atoms attached to ring carbon atoms either are not replaced or are replaced by an aliphatic hydrocarbon group which is an acyclic alkyl group of 1 to 4 carbon atoms.

14. A pharmaceutical composition according to Claim 13, in which the pyridone is a 3 - hydroxypyrid - 2 - one, 3 - hydroxypyrid - 4 - one, 3 hydroxy - 2 methylpyrid - 4 - one, 3 - hydroxy - 6 - methylpyrid - 4one or 3 - hydroxy - 2,6 - dimethylpyrid -4- one with the hydrogen atom attached to the nitrogen atom replaced by an aliphatic acyl group or by an aliphatic hydrocarbon group substituted by one or, except in the case of ionisable groups, more than one substituent selectedfrom aliphatic acyl, alkoxy, aliphatic amine, aliphatic amide, carboxy, aliphatic ester, halogen, hydroxy and sulpho groups.

15. A pharmaceutical composition according to Claim 12,13 or 14 in which the hydrogen atom attached to the nitrogen atom is replaced by an aliphatic acyl group -COR1, an aliphatic hydrocarbon group -(CH2)ncarrying a terminal substituent group -COR2, -COXIR2, -S02XR2, -XCOR2 or -XS02R,, or an aliphatic hydrocarbon group -(CH2)rn- carrying a terminal substituent group -XH, wherein R, and R2 are each an acyclic alkyl group of 1 to 4 carbon atoms, n is an integerfrom 1 to 4 and m is an integerfrom 3 to 6.

16. A pharmaceutical composition according to any of Claims 12to 15, in which the pyridone is in the form of a free compound ratherthan in the form of a saitthereof.

17. A pharmaceutical composition according to any of Claims 12 to 16, which contains a physiologically acceptable solid carrier.

18. A pharmaceutical composition according to Claim 17 in tablet or capsule form.

19. A pharmaceutical composition according to Claim 17 in suppository form.

20. A pharmaceutical composition according to any of Claims 12to 16, which haswater ora medium containing an organic solvent as the physiologically acceptable diluent and has the form of a solution, 16 suspension or emulsion.

21. A pharmaceutical composition according to Claim 20 in sterile injectable form.

22. Apharmaceuticai composition according to 5 any of Claims 12 to 21 in unit dosage form.

23. Acompound according to Claim 1 or2as described hereinbefore in any of Examples 1 to

24.

Printed in the United Kingdom for Her majesty's stationery office, 8818935, 9184, 18996. Published at the Patent office, 25 Southampton Buildings, London WC2A lAY, from which copies may be obtained.

GB 2 136 807 A 16 4 Y.