Claims
- 1. A compound of the formula ##STR12## wherein X is amino or hydroxyl; n is 2, 3 or 4
- or a salt or ester at the carboxyl group thereof.
- 2. 4-[3-(2,4-Diamino-7H-pyrrolo[2,3-d]pyrimidin--5-yl)propyl]benzoic acid or a salt or ester at the carboxyl group thereof.
- 3. The tert-butyl ester of the compound of claim 1.
- 4. The compound of claim 1 wherein X is hydroxyl.
- 5. The compound of claim 1 wherein X is amino.
Priority Claims (2)
| Number |
Date |
Country |
Kind |
| 63-71149 |
Mar 1988 |
JPX |
|
| 63-245379 |
Sep 1988 |
JPX |
|
PROCESS 1
This is a division of Ser. No. 326,901, filed Mar. 21, 1989, now U.S. 4,997,838, issued Mar. 5, 1991.
This invention relates to the novel pyrrolopyrimidine derivatives which are useful as anti-tumor agents, the production and utilization thereof.
Folic acid is a carrier of a Cl unit in a living body, derived from formic acid or formaldehyde, acting as a coenzyme in various enzymatic reactions such as those in biosynthesis of nucleic acid, in metabolism of amino acids and peptides and in generation of methane. Particularly in biosynthesis of nucleic acid, folic acid is essential for formylation in the two pathways, i.e. the purine synthetic pathway and the thymidine synthetic pathway. Usually folic acid is required to be transformed into its activated coenzyme form by reduction in two steps before it becomes biologically active. Amethopterin (methotrexate: MTX) and the related compounds are known to inhibit the reduction from dihydrofolic acid into tetrahydrofolic acid by coupling strongly with the dominant enzyme in the second step (dihydrofolic acid reductase). These drugs have been developed as antitumor drugs because they may disturb the DNA synthesis and consequently cause cell death, and are currently regarded of major clinical important. On the other hand, a novel tetrahydroaminopterin antitumor agent (5,10-dideaza-5,6,7,8-tetrahydroaminopterin: DDATHF) has been reported which, unlike the drugs described above, does not inhibit dihydrofolic acid reductase and the main mechanism of which consists in inhibition of glycinamide ribonucleotide transformylase required in the initial stage of purine biosynthesis [Journal of Medicinal Chemistry, 28, 914 (1985)].
Various studies are now being conducted on therapy for cancer, and what is expected strongly is the development of drugs which are more effective and have toxicities highly specific to cancer cells based on some new mechanism. The antitumor agent MTX the action mechanism of which consists in antagonism against folic acid, is clinically used widely, though the therapeutic effect is still unsatisfactory because it has relatively strong toxicity with little effect on solid cancer.
As the result of the inventors' researches under the circumstances described above, they have found out that novel pyrrolopyrimidine derivatives have toxicities highly specific to tumor cells and excellent antitumor effects, and completed this invention.
This invention relates to
(1) A compound of the formula (I) ##STR2## wherein the ring A is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom, an amino group or a hydroxyl group, R is a hydrogen atom, a fluorine atom, an alkyl group, an alkenyl group or an alkynyl group, --COOR.sup.1 and --COOR.sup.2 are independently carboxyl groups which may be esterified and n is an integer of 2 to 4, and R may be different in each of the n repeating units, and salts thereof,
(2) A method for production of the compounds (I) or salts thereof characterized in that a compound of the formula (II) ##STR3## wherein the ring A is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom, an amino group or a hydroxyl group, R is a hydrogen atom, a fluorine atom, an alkyl group, an alkenyl group or an alkynyl group, and n is an integer of 2 to 4, and R may be different in each of the n repeating units, a reactive derivative at the carboxyl group, or a salt thereof, and a compound of the formula ##STR4## wherein --COOR.sup.1 and --COOR.sup.2 are independently carboxyl groups which may be esterified, or a salt thereof, are allowed to react.
(3) A compound of the formula (IV) ##STR5## wherein the ring A is a pyrrole or pyrroline ring, X is an amino group or a hydroxyl group, Y is a hydrogen atom, an amino group or a hydroxyl group, R is a hydrogen atom, a fluorine atom, an alkyl group, an alkenyl group or an alkynyl group, --COOR.sup.3 is a carboxyl group which may be esterified and n is an integer of 2 to 4, and R may be different in each of the n repeating units, and salts thereof.
(4) Anti-tumor agents containing the compounds (I) or salts thereof.
When X or Y in the formulas described above is a hydroxyl group, each of the compounds (I), (II) and (IV) may exist as an equilibrium mixture of the respective tautomers. The following partial structural formulas show the sites of the structure which are subject to tautomerism, and the equilibrium between the tautomers is illustrated in the following. ##STR6##
For the convenience of description, only the hydroxyl forms and the corresponding names are described throughout this specification, but the corresponding oxo forms are always included.
There may be two or more asymmetric centers in the compounds (I) of this invention, and the absolute configuration at all of the asymmetric carbon atoms may be the S, R or S-R mixed form, except that the absolute configuration at the asymmetric carbon atom in the side chain derived from glutamic acid is always S(L). Therefore the compounds (I) may have two or more diastereomers which, if necessary, can easily be separated from each other by a routine method for separation and purification. All of the diastereomers which can be separated by such a method are included in this invention.
Alkyl groups represented by R in the formulas described above include alkyl groups having 1 to 3 carbon atom(s) each (e.g. methyl, ethyl, propyl, isopropyl groups). Alkenyl groups represented by R in the formulas described above include alkenyl groups havinig 2 to 3 carbon atom(s) each (e.g. vinyl, 1-methylvinyl, 1-propenyl, allyl, allenyl groups). Alkynyl groups represented by R in the formulas described above include alkynyl groups havinig 2 to 3 carbon atom(s) each (e.g. ethynyl, 1-propynyl, propagyl groups). Carboxyl groups in the carboxyl groups which may be esterified, represented by --COOR.sup.1, --COOR.sup.2 and --COOR.sup.3 include carboxyl groups which may be esterified by alkyl groups having 1 to 5 carbon atom(s) each, benzyl groups which may be substituted or phenyl groups which may be substituted. The alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, iso-pentyl, sec-pentyl, neo-pentyl and tert-pentyl. The benzyl groups which may be substituted include benzyl, nitrobenzyl, methoxybenzyl groups and so on. The phenyl groups which may be substituted include phenyl, nitrophenyl, methoxyphenyl groups and so on.
In the following the method for production of the compounds (I) of this invention is explained.
The compounds (I) can be obtained by acylation of glutamic acid derivatives shown by the formula (III) with carboxylic acids shown by the formula (II) or reactive derivatives thereof. The acylation may be performed, for example, by acylation of the compound (III) with the compound (II) in the presence of carbodiimide, dephenylphosphoryl azide or diethyl phosphoro cyanidate. Generally about 1 to 20 mole equivalent, preferably 1 to 5 mole equivalent of the compound (III) relative to the compound (II) is used. Generally about 1 to 25 mole equivalent, preferably about 1 to 5 mole equivalent of a carbodiimide relative to the compound (II) is used. As the carbodiimide, dicyclohexylcarbodiimide is preferable for practical use, but other carbodiimides such as diphenylcarbodiimide, di-o-tolylcarbodiimide, di-p-tolylcarbodiimide, di-tert-butylcarbodiimide, 1-cyclohexyl-3-(2-morpholinoehtyl)carbodiimide, 1-cyclohexyl-3-(4-diethylaminocyclohexyl)carbodiimide, 1-ethyl-3-(2-diethylaminopropyl)carbodiimide and 1-ethyl-3-(3-diethylaminopropyl)carbodiimide may be used. The acylation is preferably performed in the presence of a suitable solvent, and such solvents include water, alcohols (e.g. methanol, ethanol, etc.), ethers (e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme, etc.), nitriles (e.g. acetonitrile, etc.), esters (e.g. ethyl acetate, etc.), halogenated hydrocarbons (e.g. dichloromethane, chloroform, carbon tetrachloride, etc.), aromatic hydrocarbons (e.g. benzene, toluene, xylene, etc.), acetone, nitromethane, pyridine, dimethylsulfoxide, dimethylformamide, hexamethylphospholamide, sulfolane, and the suitable mixtures of two or more of these solvents. The reaction is allowed to proceed generally at a pH ranging from 2 to 14, preferably at a pH ranging from about 6 to 9, at a temperature ranging from about -10.degree. C. to the boiling point of the solvent used (up to about 100.degree. C.), preferably at a temperature ranging from about 0 to 50.degree. C., for about 1 to 100 hours. The pH of the reaction mixture is adjusted, if necessary, by addition of an acid (e.g. hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, acetic acid, etc.), a base (e.g. sodium alcoholate such as sodium methylate and sodium ethylate, hydroxides of alkali metals or of alkali earth metals such as sodium hydroxide, potassium hydroxide, lithium hydroxide, barium hydroxide, carbonates or bicarbonates of alkali metals or of alkali earth metals such as sodium carbonate, potassium carbonate, barium carbonate, calcium carbonate and sodium biccarbonate, amines such as trimethylamine, triethylamine, triethanolamine and pyridine), or a buffer (e.g. phosphate buffer, borate buffer, acetate buffer, etc.). The reaction can proceed more advantageously in the presence of a catalyst which promotes acylation. Such catylysts include base catalysts and acid catalysts. The base catalysts include tertiary amines (e.g. aliphatic tertiary amines such as triethylamine; aromatic tertiary amines such as pyridine, .alpha.-, .beta.- or .gamma.-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and diethylaniline), and such acid catalysts include Lewis acids [e.g. anhydrous zinc chloride, anhydrous aluminum chloride (AlCl.sub.3), anhydrous ferric chloride, titanium tetrachloride (TiCl.sub.4), tin tetrachloride (SnCl.sub.4), antimony pentachloride, cobalt chloride, cupric chloride, boron trifluoride ethyl ether complex, etc.]. Among the catalysts described above, 4-dimethylaminopyridine or 4-(1-pyrrolidinyl)pyridine is preferable in many cases. The suitable amount of the catalyst is such that is enough to promote the acylation, being generally about 0.01 to 10 mole equivalent, preferably about 0.1 to 1 mole equivalent relative to the compound (II). The reactive derivatives of carboxylic acids obtained by the reaction at the carboxyl group, used for the acylation include acid halides (e.g. fluoride, chloride, bromide, iodide), acid anhydrides (e.g. iodoacetic acid anhydride, isobutyric acid anhydride), mixed acid anhydrides with monoalkylcarbonic acid esters (e.g. mono-methylcarbonic acid ester, monoethylcarbonic acid ester, monopropylcarbonic ester, mono-iso-propylcarbonic acid ester, monobutylcarbonic acid ester, mono-iso-butylcarbonic acid ester, mono-sec-butylcarbonic acid ester, mono-tert-butylcarbonic acid ester), active esters (e.g. cyanomethyl ester, carboethoxymethyl ester, methoxymethyl ester, phenyl ester, o-nitrophenyl ester, p-nitrophenyl ester, p-carbomethoxyphenyl ester, p-cyanophenyl ester, thiophenyl ester), acid azides, mixed acid anhydrides with phosphoric acid diesters (e.g. dimethyl phosphate, diethyl phosphate, dibenzylphosphate, diphenylphosphate), and mixed acid anhydrides with phosphorous acid diesters (e.g. dimethyl phosphite, diethyl phosphite, dibenzyl phosphite, diphenyl phosphite), of the carboxylic acid (II). For acylation with such a reactive derivative, the solvent, the catalyst and the reaction temperature are the same as for acylation in the presence of the carbodiimide described above.
For production of the compound (I-1) in which --COOR.sup.1 and --COOR.sup.2 in the formula of the compound (I) are carboxyl groups, it is desirable that the compound in which --COOR.sup.1 and --COOR.sup.2 in the formula of the compound (III) are esterified carboxyl groups is allowed to react with the compound (II) followed by deesterification by per se known degradation or catalytic reduction. Such degradation can be performed by hydrolysis under basic conditions (method A), hydrolysis under acidic conditions (method B-1) or hydrolysis under acidic nonaqueous conditions (method B-2). Bases used in the method A include metal alkoxides such as sodium methoxide, sodium ethoxide, sodium butoxide and potassium butoxide, metal hydroxides such as sodium hydroxide, potassium hydroxide, lithium hydroxide and barium hydroxide, and amines such as ammonia, triethylamine and pyridine. Acids used in the method B-1 include mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid, and organic acids such as trifluoroacetic acid, trichloroacetic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and camphorsulfonic acid. Catalysts used in the method B-2 include mineral acids such as hydrogen chloride, hydrogen bromide, perchloric acid, sulfuric acid, nitric acid and phosphoric acid, organic acids such as trifluoroacetic acid, trichloroacetic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and camphorsulfonic acid, and Lewis acids such as anhydrous zinc chloride, anhydrous aluminum chloride (AlCl.sub.3), anhydrous ferric chloride, titanium tetrachloride (TiCl.sub.4), tin tetrachloride (SnCl.sub.4), antimony pentachloride, cobalt chloride, cupric chloride and boron trifuluoride ethyl ether complex. Degradation is performed in a suitable solvent at a temperature ranging from 0.degree. C. to the boiling point of the solvent, preferably at 10.degree. to 80.degree. C., for 30 minutes to 2 days. The solvent used for the reaction by the method A or by the method B-1 may be water, methanol, ethanol, propanol, butanol, ethyleneglycol, methoxyethanol, ethoxyethanol, tetrahydrofuran, dioxane, monoglyme, diglyme, pyridine, dimethylformamide, dimethylsulfoxide or sulfolane, or a suitable mixture of two or more of these solvents; the solvent used for the reaction by the method B-2 may be ethyl acetate, dimetyl ether, diethyl ether, tetrahydrofuan, dioxane, monoglyme, diglyme, dichloromethane, chloroform, carbon tetrachloride, acetonitrile, benzene, toluene, xylene, nitromethane or pyridine, or a suitable mixture of two or more of these solvents. The catalytic reduction (method C) is performed in a suitable solvent at a temperature ranging from about -40.degree. C. to the boiling point of the solvent used, preferably at about 0.degree. to 50.degree. C. The solvents used include water, alcohols (e.g. methanol, ethanol, propanol, iso-propanol, butylalcohol, sec-butylalcohol, tert-butylalcohol, ethyleneglycol, methoxyethanol, ethoxyethanol), acetic acid esters (e.g. methyl acetate, ethyl acetate), ethers (e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme, aromatic hydrocarbons (e.g. benzene, toluene, xylene), pyridine, dimethylformamide and suitable mixtures of two or more of these solvents. Catalysts for the catalytic reaction include palladium, platinum, rhodium and Raney nickel. Addition of a trace amount of acetic acid, trifluoroacetic acid, hydrochloric acid or sulfuric acid can allow the reaction to proceed advantageously.
The method for production of the compound (I-1) is selected according to the nature of --COOR.sup.1 and --COOR.sup.2 ; when --COOR.sup.1 and --COOR.sup.2 are carboxyl groups esterified with methyl, ethyl, propyl, butyl, sec-butyl, phenyl or substituted phenyl group, the method A or the method B-1 is applied advantageously; when --COOR.sup.1 and --COOR.sup.2 are carboxyl groups esterified with iso-propyl or tert-butyl group, the method B-2 is applied advantageously; and when --COOR.sup.1 and --COOR.sup.2 are carboxyl groups esterified with benzyl or a substituted benzyl group, the method B-1 or the method C is applied advantageously. When --COOR.sup.1 and --COOR.sup.2 are different from each other, the methods A, B-1, B-2 and C may be combined appropriately.
In the following the method for production of the starting compound (II) is explained.
The compound (II) wherein the ring A is a pyrrole ring, can be produced, for example, by the following processes. ##STR7##
In the reaction formulas described above, X, Y and R.sup.3 are the same as described before; R.sup.a, R.sup.b and R.sup.c are independently a hydrogen atom, a fluorine atom or an alkyl group (the same as those represented by R described before); R.sup.4 is a cyano group or an esterified carboxyl group represented by the formula --COOR.sup.5 ; A is a hydrogen atom or a halogen atom (e.g. fluorine atom, chlorine atom, bromine atom, iodine atom); B is a halogen atom (e.g. chlorine atom, bromine atom, iodine atom) or an eliminable group which may be easily derived from hydroxy group (e.g. methanesulfonyloxy group, benzenesulfonyloxy group, p-toluenesulfonyloxy group, trifluoromethanesulfonyloxy group); and m is 0, 1 or 2. R.sup.5 in the esterified carboxyl group represented by the formula --COOR.sup.5 is exemplified by an alkyl group having 1 to 4 carbon atom(s) (e.g. methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, etc.), phenyl or substituted phenyl group (p-nitrophenyl, p-methoxyphenyl, etc.), and benzyl or substituted benzyl (e.g. p-nitrobenzyl, p-methoxybenzyl, etc.).
The compound (V) may be dehydrogenated on the possible position between the two adjacent carbons and form an unsaturated bond.
In the following the reaction processes described above are explained in detail.
The compound (V) and the compound (VI) are subjected to condensation and the resulting product is subjected to reduction to give the compound (VII).
For the condensation, a known reaction (e.g. aldol reaction, Reformatsky reaction, Wittig reaction, etc.) is employable, and for the reduction, usually a catalytic reduction under hydrogen atmosphere in the presence of a catalyst (e.g. nickel, palladium, platinum, rhodium) is advantageously employed.
In the condensation by aldol reaction, the employable base catalysts include metal hyrdoxides such as sodium hydroxide, potassium hydroxide, lithium hydroxide and barium hydroxide, metal alkoxides such as sodium methoxide, sodium ethoxide and potassium tert-butoxide, metal amides such as sodium amide and lithium diisopropylamide, metal hydrides such as sodium hydride and potassium hydride, organic metal compounds such as phenyllithium and butyllithium and amines such as triethylamine, pyridine, .alpha.-, .beta.- or .gamma.-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and diethylaniline; the employable acid catalysts include mineral acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and boric acid, and organic acids such as oxalic acid, tartaric acid, acetic acid, trifluoroacetic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and camphorsulfonic acid. The condensation can be conducted according to the known method [Ei-Ichi Negishi, Organometallics in Organic Synthesis, vol. 1, John Wiley & Sons, New York, Chichester, Brisbane, Toronto (1980)] which comprises converting a ketone form into the silylenolether form which is then subjected to condensation with an aldehyde or an equivalent in the presence of a Lewis acid [e.g. anhydrous zinc chloride, anhydrous aluminum chloride (AlCl.sub.3), anhydrous ferric chloride, titanium tetrachloride (TiCl.sub.4), tin tetrachloride (SnCl.sub.4), antimony pentachloride, cobalt chloride, cupric chloride, boron trifluoride ethyl ether complex, etc.], or converting a ketone form into the enolate by treating the ketone form with a metal triflate (e.g. dialkyl boron tin (II) triflate) in the presence of amines (e.g. triethylamine, pyridine, .alpha.-, .beta.- or .gamma.-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline, diethylaniline) followed by subjecting the enolate to condensation with an aldehyde or an equivalent. The condensation is conducted in a suitable solvent at a temperature ranging from -100.degree. C. to the boiling point of the solvent, preferably ranging from -78.degree. to 100.degree. C., for 1 minute to 3 days. Solvents employable for the reaction include water, liquid ammonia, alcohols (e.g. methanol, ethanol, propanol, isopropanol, butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, ethylene glycol, methoxyethanol, ethoxyethanol), ethers (e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme), halogenated hydrocarbons (e.g. dichloromethane, chloroform, carbon tetrachloride), aliphatic hydrocarbons (e.g. pentane, hexane, heptane), aromatic hydrocarbons (e.g. benzene, toluene, xylene), dimethylformamide, dimethylsulfoxide, hexamethylphospholamide, sulfolane, and the suitable mixtures thereof. In the condensation by Wittig reaction, the employable reagents include metal hydroxides such as sodium hydroxide, potassium hydroxide, lithium hydroxide and barium hydroxide, metal alkoxides such as sodium methoxide, sodium ethoxide and potassium tert-butoxide, metal amides such as sodium amide and lithium diisopropylamide, metal hydrides such as sodium hydride and potassium hydride, organic metal compounds such as phenyllithium and butyllithium, and amines such as triethylamine, pyridine, .alpha. -, .beta.- or .gamma.-picoline, 2,6-lutidine, 4-dimethylaminopyridine, 4-(1-pyrrolidinyl)pyridine, dimethylaniline and diethylaniline. The reaction is conducted in a suitable solvent at a temperature ranging from -20.degree. C. to the boiling point of the solvent used, preferably ranging from 0.degree. to 150.degree. C., for 1 minute to 10 days. The solvents employable for the reaction include liquid ammonia, alcohols (e.g. methanol, ethanol, propanol, isopropanol, butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, ethylene glycol, mehoxyethanol, ethoxyethanol), ethers (e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme, aliphatic hydrocarbons (e.g. pentane, hexane, heptane), aromatic hydrocarbons (e.g. benzene, toluene, xylene), dimethylformamide, dimethylsulfoxide, hexamethylphospholamide, sulfolane and the suitable mixtures thereof.
The condensation can also be conducted by using a Reformatsky reaction. The reagents employable for the Reformatsky reaction include zinc, magnesium, aluminum and tin, and the reaction is conducted in a suitable solvent at a temperature ranging from -20.degree. C. to the boiling point of the solvent used, preferably ranging from 0.degree. to 150.degree. C., for 30 minutes to 3 days. The solvents employable for the reaction include ethers (e.g. dimethyl ether, diethyl ether, tetrahydrofuran, dioxane, monoglyme, diglyme), aliphatic hydrocarbons (e.g. pentane, hexane, heptane), aromatic hydrocarbons (e.g. benzene, toluene, xylene) and the suitable mixtures thereof.
The reaction conditions for the catalytic reduction are the same as those for the deesterification at the --COOR.sup.1 and --COOR.sup.2 of the compound (III) (method C).
The starting materials (V) and (VI) can be obtained easily according to the known methods described in the literature. [B. Neises et al., Angew. Chem. Int. Ed. Engl., 17, 522 (1978)].
This is the process whereby an eliminable functional group B is introduced into the active methylene (the .alpha.-position of the carbonic acid ester) of the compound (VII): it can be conducted easily by using known reagents according to a per se known method.
The compound (VIII) obtained in the Process 2 is subjected to condensation with malononitrile or a cyanoacetic acid ester [NC-CH.sub.2 COOR.sup.5 ; R.sup.5 is the same as described above] under a basic condition, to give the compound (IX). The employable bases, solvents and reaction conditions are in accordance with the known methods.
The compound (IX), when treated with guanidine, can react at the cyano group or the ester residue followed by ring closure to form newly a pyrrolopyrimidine ring. Ring closure under a basic condition allows the reaction to proceed advantageously. The employable bases include metal alkoxides such as sodium methoxide, sodium ethoxide and potassium tert-butoxide. The employable solvents for the reaction include methanol, ethanol, propanol, tert-butyl alcohol, dimethylsulfoxide and hexamethylphospholamide. The reaction temperature ranges from 0.degree. to 150.degree. C., preferably from 20.degree. to 100.degree. C. The reaction time ranges from 1 to 48 hours.
The compound (IV-1: Y=NH.sub.2 or OH) obtained in the Process 4 can be converted into the compound (II-1: Y=NH.sub.2 or OH) by subjecting the ester residue [--COOR.sup.3 ] to the deesterification used in the preparation of the compound (I-1).
The compound (II-1: Y=OH) obtained in the Process 5 is subjected to reduction to give the compound (II-2: Y=H). The conditions for the reduction are per se known, and reduction by a metal hydride (e.g. borane, alane or ate complexes thereof) is employed advantageously.
The Process 5 and the Process 6 may be conducted in the reverse order. Namely, in the Process 7 the compound (IV-1: Y=OH) is subjected to reduction similar to that in the Process 6 to give the compound (IV-2: Y=H), which is then subjected to deesterification in the Process 8 in a similar manner as in the Process 5 to give the compound (II-2: Y=H). Either the deesterification or the reduction can be selected to be conducted in advance to the other according to the nature of the substituents in the compound (IV-1: Y=OH).
In the above Processes 6 and 8, the mixture containing the compounds (II-2) and (II-2') or the compounds (IV-2) and (IV-2') may be separated, or each of the compounds (II-2) and (II-2') or each of the compounds (IV-2) and (IV-2') is synthesized predominantly by selective reduction.
Among the compounds (II), those represented by the formula (II-3: X=OH) ##STR8## wherein R and n means the same as described before, can be obtained also by the following processes. ##STR9##
In the Processes described above, R, Ra, Y and n means the same as described before and Z means the formula RCH.sub.2 CO-- wherein R means the same as described above, the formula ##STR10## wherein L is phenyl, butyl or cyclohexyl, and R and n means the same as described above, or the formula ##STR11## wherein M is ethyl or phenyl, and R and n means the same as described above. It is preferable that Y is hydrogen.
In the following these Processes are explained.
The compound (X) [T. Kondo et al., Chemistry Letters, 419 (1983)] and a para-substituted benzoic acid ester derivative (XI) are subjected to condensation (aldol reaction, Wittig reaction) followed by catalytic reduction under hydrogen atmosphere, to give the compound (XII). For the condensation are applicable the reaction conditions, the reaction solvents, the reaction temperatures and the reagents used in the Process 1. For the catalytic reduction under hydrogen atmosphere are applicable the conditions used in the deesterification of --COOR.sup.1 and --COOR.sup.2 of the compound (III).
Treatment of the compound (XII) under acidic conditions can eliminate the protection of the isopropyloxymethyl group at the 3-position to give the compound (XIII). The conditions, solvents and temperatures used in deesterification of --COOR.sup.1 and --COOR.sup.2 of the compound (III) (the method B-1 and the method B-2) are employable for the reaction.
The compound (XIII) obtained in the Process 10 is subjected to dehydrogenation by a per se known method, to be easily converted into the compound (IV-3: Y=H).
The compound (IV-3: Y=H) obtained in the Process 11 can be converted into the compound (II-3) by deesterification. The conditions, solvents and temperatures described in detail for the deesterification of --COOR.sup.1 and --COOR.sup.2 of the compound (III) (the methods A, B-1, B-2 and C) are employable for the reaction. The processes 10 to 12 may be conducted in any order with the formation of the respective products, and finally the desired compound (II-3) is obtained. The order is determined suitably according to the nature of the substituents of the compounds (XII), (XIII) and (IV-3). The compound (II-3) thus obtained can be converted, if necessary, into the compound (II-2) by a known substituent-converting reaction on the pyrimidine ring reported in the literature. [Protein Nucleic acid Enzyme Extra Issue, Chemical synthesis of nucleic acids, Kyoritsu Shuppan (1968)].
The compounds other than the compound (II-3), wherein X is hydroxyl can be also converted into the corresponding compounds wherein X is amino by the above-mentioned substituent-converting reaction.
The reactions, reagents and reaction conditions used in the Processes 1 to 12 and in the production of the starting compounds (V) and (XIII) are known and explained in detail in the following literature. [J. F. W. Mcomine, Protective Groups in Organic Chemistry, Plenum Press, London and New York (1973)], [Pine, Hendrikson, Hammond, Organic Chemistry (4th edition) [I]-[II], Hirokawa Shoten (1982)], and [M. Fieser and L. Fieser, Reagents for Organic Synthesis vol. 1-10, Wiley-Interscience, New York, London, Sydney and Toronto (1969-1982)].
The intermediates of the compounds of this invention and the compounds (I) of this invention can be isolated from the reaction mixtures by the conventional means for separation and purification, such as concentration, extraction with solvent, chromatography and recrystallization.
The compounds (I), (II) and (IV) of this invention may form salts. Such salts are produced by the known methods, and exemplified by the salts of pharmaceutically acceptable bases or acids and quaternary salts. Salts of bases include salts of alkali metals, alkali earth metals, non-toxic metals, ammonium and substituted ammonium, such as sodium, potassium, lithium, calcium, magnesium, aluminum, zinc, ammonium, trimethylammonium, triethylammonium, triethanolammonium, pyridinium and substituted pyridinium. Salts of acids include salts of mineral acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and boric acid, and salts of organic acids such as oxalic acid, tartaric acid, acetic acid, trifluoroacetic acid, methanesulfonic acid and camphorsulfonic acid. Quaternary salts include salts of methyl bromide, methyl iodide, methyl methanesulfonate, methyl benzensulfonate and methyl p-toluenesulfonate. Also, the compounds (I), (II) and (IV) may form zwitterion.
As the compounds (I) of this invention, the following compounds are exemplified:
The compounds (I) of this invention show excellent antitumor effects in mouse tumor cell strains (P388, L1210, L5178Y, B16 melanoma, MethA, Lewis Lung Carcinoma, S180 sarcoma, Ehrlich Carcinoma, Colon38) and human tumor cell strains (HL60, KB, Lu65), decrease the tumors carried by warm-blooded animals [e.g. melanoma, sarcoma, mastocytoma, carcinoma, neoplasia, etc.] and prolong the life-span of tumor-carrying warm-blooded animals.
In the following are described the results indicating the pharmaceutical effects of the compounds (I) of this invention.
The cell growth inhibiting effect (IC.sub.50) of the compounds obtained in the Working Examples described below in KB cells was determined by the following method.
Human nasopharyngeal cancer KB cells (1.times.10.sup.4 cells/ml) prepared according to a conventional method were inoculated into each well of the 96-microwell plate (0.1 ml in a well) and subjected to standing culture at 37.degree. C. under 5% CO.sub.2 for 24 hours. To this was added a solution of one of the compounds obtained in the Working Examples in 10% MEM (Nissui Pharmaceutical Co. Ltd.), and subjected again to standing culture at 37.degree. C. under 5% CO.sub.2 for 72 hours. Then the culture was pipetted out, and another 0.1 ml of the solution of MTT (Dojindo Laboratories) in 10% MEM (1.0 mg/ml) was added and incubated at 37.degree. C. for 4 hours. Then 0.1 ml of the 10% SDS solution (Wako Pure Chemicals) was added and incubated at 37.degree. C. for further 24 hours. The absorbance at 590 nm was measured and the IC.sub.50 value of the compound was defined as the concentration of the compound required to decrease the number of cells in the untreated control group by 50%. The results obtained are shown in Table 1.
In addition, the following are described the results indicating the pharmaceutical effects of the compounds (I) of this invention.
The cell growth inhibiting effect (IC50) of the compound obtained in the Working Example 14 described below in HL-60 and HEL cells was determined by the following method.
As shown by the above-mentioned results, the compounds (I) are excellent in inhibition of cell growth of KB and HL-60, while they do not exert a toxicity against HEL. The compounds (I) of this invention and the salts thereof are of low-toxicity, having remarkable antitumor effect. Therefore, the preparations containing the compound (I) or salts thereof can be employed as antitumor agents for the treatment of tumors in warm-blooded animals, particularly mammals (e.g. mouse, rat, cat, dog, rabbit, etc.).
The compounds (I) and salts thereof, when used as antitumor agents, can be administered orally and parenterally as they are or in the forms of powders, granules, tablets, capsules, suppositories and injections, which are prepared according to the conventional methods using pharmaceutically acceptable excipients, vehicles, and diluents. The dose varies according to the animals, diseases, symptoms, compounds and administration routes; for example, the daily dose is about 2.0 to 100 mg of a compound of this invention per kg of body weight of a warm-blooded animal described above for oral administration, and about 1.0 to 50 mg/kg for parenteral administration. Injections may be administered intramuscularly, intraperitoneally, subsutaneously or intravenously.
The preparations are produced by the per se known processes. For the above-mentioned oral preparations, for example, tablets are produced by suitable combination with a binder (e.g. hydroxypropylcellulose, hydroxypropylmethylcellulose, macrogol, etc.), a disintegrator (e.g. starch, calcium carboxylmethylcellulose, etc.) and a lubricant (e.g. magnesium stearate, talc, etc.).
As parenteral preparations, for example, injections are produced by suitable combination with an agent to provide isotonicity (e.g. glucose, D-sorbitol, D-mannitol, sodium chloride, etc.), an antiseptic (e.g. benzyl alcohol, chlorobutanol, methyl p-hyrdoxybenzoate, propyl p-hydroxybenzoate, etc.) and a buffer (e.g. phosphate buffer, sodium acetate buffer, etc.).
An example process for production of tablets comprises mixing about 1.0 to 25 mg of the compound of this invention, 100 to 500 mg of lactose, about 50 to 100 mg of corn starch and about 5 to 20 mg of hydroxypropylcellulose for preparation of a tablet by a conventional means, granulating, mixing with corn starch and magnesium stearate and tabletting, so that tablets each weighing about 100 to 500 mg with the diameter of about 3 to 10 mm are obtained. The tablets may be coated with a mixture of acetone and ethanol, the mixture containing hydroxypropylmethylcellulose phthalate (about 10 to 20 mg per tablet) and castor oil (0.5 to 2 mg) at a concentration of about 5 to 10%, to give enteric coated tablets.
An example process for injectable preparation comprises dissolving about 2.0 to 50 mg of a sodium salt of the compound of this invention in about 2 ml of physiological saline for preparation of an ampoule, sealing the resultant solution in an ampoule and sterilizing the ampoule at 110.degree. C. for about 30 minutes or adding about 2 ml of sterile distilled water containing about 10 to 40 mg of mannitol or sorbitol into the ampoule, freeze-drying and sealing the ampoule. For use of the freeze-dried compound for subcutaneous, intravneous or intramuscular injection, the ampoule is opened and the content is dissolved in, for example, physiological saline so that the concentration of the compound may be about 1.0 to 25 mg/ml.
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Divisions (1)
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Number |
Date |
Country |
| Parent |
326901 |
Mar 1989 |
|
Patent Grant
5106974
