KR100382778B1 - Compounds Useful as Antiproliferative and GARFT Inhibitors - Google Patents

Abstract

The present invention relates to compounds represented by formula (I) in equilibrium with 4-hydroxy tautomers and pharmaceutically acceptable salts thereof. These compounds are useful as glycineamide ribonucleotide formyl transferase (GARFT) inhibitors or antiproliferative agents. The present invention also relates to methods of applying these compounds as pharmaceutical compositions and GARFT inhibitors or antiproliferative agents. The invention also relates to intermediate compounds useful in the preparation of these compounds.

Description

Compounds useful as antiproliferative and GARFT inhibitors

The present invention relates to a compound represented by the following formula (I) to inhibit the activity of a glycineamide ribonucleotide formyl transferase (hereinafter referred to as "GARFT") enzyme. In addition, the present invention provides an intermediate compound used in the preparation of the compound represented by the following formula (I), a pharmaceutical composition containing the same, the use of the same for the purpose of inhibiting GARFT, and microorganisms such as bacteria, yeast, mold and the like It relates to a use for the purpose of inhibiting cell growth and proliferation. In addition, the compounds of the present invention have anti-cancer, anti-inflammatory, anti-psoriasis and / or immunosuppressive effects. The present invention also relates to a process for the preparation of these compounds.

Large classes of anti-inflammatory agents include anti-metabolic compounds, and one of the subclasses of anti-metabolic compounds is antifolates or antifoles, which are antagonists of vitamin folic acid. to be. In general, antifolates are very similar to the structure of folic acid and contain the p-benzoyl glutamate portion of folic acid. The glutamate portion of folic acid has a double negative charge at physiological pH. Thus, these compounds and analogs exert a metabolic effect by activating an active transport system that requires energy as it passes through the cell membrane.

GARFT acts as a folate dependent enzyme in the biosynthesis of new de novo purine. Such biosynthetic processes have limitations in cell division and cell proliferation. Thus, blocking of such biosynthetic processes is known to have a proliferation inhibitory effect, in particular an antitumor effect. Thus, many folate analogues have been synthesized for GARFT inhibition, and much research has been conducted regarding GARFT inhibition. 5,10-Dideazatetrahydrofolic acid (DDATHF) has been reported to exhibit antitumor activity as a potent inhibitor of GARFT [FM Muggia, "Folate antimetabolites inhibitor to de novo purine synthesis," New Drugs, Concepts and Results in Cancer Cancer Chemotherapy , Kluwer Academic Publishers, Boston (1992), 65-87].

Recent studies have shown that heterocyclic glutamic acid derivatives are useful as antiproliferative or GARFT inhibitors (international patent application PCT / US94 / 00418, filed Jan. 18, 1994). Continuing their work, the researchers developed compounds useful as antiproliferative and GARFT inhibitors.

The present invention relates to a compound represented by the following formula (I). These compounds inhibit the activity of glycinamide ribonucleotide formyl transferase (hereinafter referred to as "GARFT") and are effective for inhibiting cell growth and proliferation of microorganisms such as bacteria, yeast, and fungi and higher organisms. . The present invention also relates to pharmaceutical compositions containing these compounds or their appropriate salts and to the use of these compounds as inhibitors of the GARFT enzyme.

As described above, the compound of the present invention has a proliferation inhibitory effect expressed by antitumor activity. The compounds of the present invention may be active as such, or may be precursors that can be converted into active substances in vivo. Preferred compounds of the invention show activity in the inhibition of GARFT enzymes. Particularly preferred compounds are active in inhibiting the growth of a mouse leukemia cell line, ie L1210 cell line, which can grow in tissue culture. In addition, the compounds of the present invention can also be active in inhibiting the growth of bacteria such as Escherichia coli gram negative bacteria that can grow in culture.

Pharmaceutically acceptable salts thereof, including the compounds according to the invention, may be contained in conventional dosage forms such as capsules, tablets or injections. Solid or liquid pharmaceutically acceptable carriers, diluents or excipients may also be applied.

Starch, lactose, calcium sulfate dihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate and stearic acid may be used as a solid carrier, and syrup, Peanut oil, olive oil, saline and water can be used. As a carrier or diluent, for example, glyceryl monostearate or glyceryl distearate alone or certain prolonged release materials used in combination with wax may be included. If liquid carriers are used, they are formulated in the form of syrups, exciters, emulsions, soft gelatin capsules, sterile injectable solutions (such as solutions), non-aqueous suspensions or aqueous suspensions.

Pharmaceutical formulations are by conventional techniques well known to pharmaceutical chemists, i.e. by mixing, granulating and pressing components during manufacture for tablets, or by mixing, filling and dissolving the components. . In this case, the content of the effective ingredient is adjusted according to the dosage form, for example, oral, parenteral, topical, intravenous, intranasal, intrabronchial, intraocular, intraoral and rectal administration. do.

The pharmaceutical composition of the present invention may contain one or two or more other compounds having antitumor activity. Another compound having antitumor activity may include a mitotic inhibitor, such as vinblastine; Alkylating agents; Dihydrofolate reductase inhibitors or TS inhibitors; Antimetabolites such as 5-fluorouracil, cytosinearabinoside, intercalating antibiotics such as adriamycin, bleomycin; Enzymes such as asparaginase; Topical isomerase inhibitors such as etoposide; Biological response modifiers, for example interferon. In addition, the compounds of the present invention can be used together with one or two or more commonly known anti-proliferative or GARFT inhibitors, for example, International Patent Publication No. WO 94/13295 (published on June 23, 1994) or international patent There are compounds described in publication WO 92/05153 (published April 4, 1992).

In addition, the pharmaceutical compositions of the present invention are antibacterial agents (antibacterial agents), antifugal agents (antifugal agents), antiparasitic agents, antiviral agents (antiviral agents), antipsoriatic agents and anticoccidial agents It may also contain another compound alone or more selected from.

Antibacterial agents include sulfonamides such as sulfamethoxazole, sulfadiazine, sulfameter or sulfadoxine; Dihydrofolic reductase inhibitors such as trimethoprim, bromodiaprim or trimmetrexate; Penicillins; Cephalosporins; Quinolones carboxylic acids and their conjugated isothiazol analogs (fused isothiazolo analogos).

In addition, the present invention includes a therapeutic process of inhibiting cell growth and proliferation of higher organisms or microorganisms by administering an effective amount of a compound according to the present invention to a host, and in particular, the compound of the present invention is a human host. It is useful for the treatment of mammalian hosts such as) and avian hosts. A particularly preferred course of treatment is the administration of an effective amount of a compound of the invention to a host for GARFT inhibition.

Accordingly, many proliferative inhibitors or pharmaceutically acceptable salts thereof can be applied in the treatment of the present invention, and these compounds can be applied to pharmaceutically acceptable pharmaceutical compositions consisting of the diluents or carriers set forth above.

Preferred dosages of the compounds are dosage units consisting of an effective amount of the active compound. The term "effective amount" is an amount sufficient to inhibit folate metabolism, which is sufficient to obtain a therapeutic effect by administering one or more drug dosage units.

The daily dosage applicable to a vertebrate host is at least 1 g of active compound per kg of host, preferably at least 0.5 g, more preferably at 100 mg, and most preferably at most about 50 mg per kg of host weight. . Drugs whose dosage has been determined are administered to a warm-blooded animal or mammal, for example a human patient who needs to treat a disease mediated by inhibition of plate metabolic pathways It depends on the method of treatment. Conventional treatments include topical treatments, such as ointments or creams; Oral therapy; Rectal therapy, eg suppositories; Parenteral by injection; There is a continuous intravenous, intranasal, bronchial, intraocular or intraocular injection method.

The compound according to the present invention may have one or more effects such as anti-proliferative effect, antibacterial effect, antiparasitic effect, antiviral effect, anti-psoriasis effect, antiprotozoal effect, anti-worm effect, anti-inflammatory effect, immune effect or antimicrobial effect. Indicates. In particular, the compounds of the present invention are useful for treating antitumor effects of spinal hosts whose tumors are dormant.

Referring to the present invention in more detail as follows.

In particular, the present invention relates to a compound represented by the following formula (I).

[Formula I]

In the above formula:

A represents an oxygen atom, a sulfur atom or Se;

X is substituted or unsubstituted C ₁ to C ₃ alkyl group, substituted or unsubstituted C ₂ to C ₃ alkenyl group, substituted or unsubstituted C ₂ to C ₃ alkynyl group, substituted or unsubstituted amino group, sulfur atom or oxygen Represents an atom;

Y represents an oxygen atom, a sulfur atom or NH;

B represents a hydrogen atom or a halogen atom (F, Cl, Br or I);

C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ to C ₆ alkyl group; And

R ₁ and R ₂ each independently represent a moiety, for example, a C ₁ to C ₆ alkyl group, which is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group.

The present invention also relates to pharmaceutically acceptable salts of the compounds represented by formula (I).

In addition, the compound represented by the formula (I) according to the present invention is a 4-oxo type, which is explained by the detailed description of the invention. The oxo group is in an autotomic equilibrium with the 4-hydroxy group. Thus, the "compound represented by Formula I" may be described as structurally including 4-oxo and automeric 4-hydroxy forms. Accordingly, the present invention relates to pharmaceutically acceptable salts of 4-hydroxytautomers of the compounds represented by the above formula (I).

Preferably X or C is C ₁ to C ₆ alkyl, C ₂ to C ₆ alkenyl, C ₂ to C ₆ alkynyl, acyl, halogen, amino, hydroxy, nitro, mercapto, monocyclic carbocycle , Monocyclic heterocycles, nonfused polycyclic carbocycles, nonfused polycyclic heterocycles, hydroxy C ₁ to C ₆ alkyl, and C ₁ to C ₆ alkoxy C ₁ to C ₆ is substituted with a substituent selected from alkyl.

Preferably X is CH ₂ , CH ₂ CH ₃ , NH, oxygen atom, sulfur atom, CH (CH ₂ OH) or NCH ₃ . Y is preferably a sulfur atom or an oxygen atom. Preferably, B is a hydrogen atom. Preferably, C is a hydrogen atom or CH ₃ . Preferably, R ₁ and R ₂ are each independently a hydrogen atom, C ₁ to C ₆ alkyl, hydroxyalkyl, alkylaryl or aralkyl, most preferably a hydrogen atom or C ₁ to C ₂ alkyl to be. Especially preferred are A and Y each being a sulfur atom and X is CH ₂ .

Compounds represented by Formula I are useful as GARFT inhibitors. Among the compounds represented by the above formula (I), R ₁ and R ₂ are each hydrogen atoms, and have excellent activity as an antitumor or antiproliferative agent. Among the compounds represented by Formula I, R ₁ and R ₂ each have a moiety (preferably an ethyl group) that forms a hydrolysable ester group together with a carboxyl group are useful for preparing a compound of free glutamic acid form. It is a novel intermediate compound, which is hydrolyzed in vivo to act as drug prodrugs.

The present invention also includes pharmaceutically acceptable salts, for example alkali metals, alkali metals, other non-toxic metals of the glutamic acid compounds according to the invention, ammonium salts of glutamic acid and substituted ammonium. Specifically, it is sodium, potassium, lithium, calcium, magnesium, pyridinium and substituted pyridinium salts of free acid compounds according to the present invention.

Hereinafter, the preparation process of the compound represented by Chemical Formula I according to the present invention will be described in detail.

In the preparation of the compound represented by the formula (I) wherein X is CH ₂ , the compound represented by the following formula (II) is useful as a starting material:

[Formula II]

Wherein: B, C and Y are each as defined in formula (I) above; D represents Cl, Br or I; And R ₆ represents a moiety that is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group. Preferably, when D is bromo or iodo. Preferably a hydrogen atom, C ₁ ~ C ₆ alkyl, hydroxy, and when the alkyl, aryl or aralkyl, in the case of more preferably a hydrogen atom or C ₁ ~ C ₂ alkyl.

The compound represented by Formula II is reacted with the compound represented by Formula III:

[Formula III]

In the formula: R ₃ represents a substituted or unsubstituted C ₁ to C ₆ alkyl group or a trisubstituted silyl group. Preferably, R ₃ is CH ₂ OH or trimethylsilyl.

The compounds represented by formulas (II) and (III) may partially dissolve at least one of the reactants in the presence of a suitable transition metal catalyst (preferably palladium or nickel), a non-nucleophilic cobase (preferably substituted amine). Reaction in a solvent produces a compound represented by the following formula (IV):

[Formula IV]

Wherein: B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above; And R ₃ is as defined in Formula III above.

When R ₃ is a trimethylsilyl group, the silyl group may be prepared using a nucleophilic base (eg methanolic or ethanol potassium carbonate) or a fluoride salt (eg potassium fluoride, cesium fluoride or tetrabutylammonium fluoride) The reactants are reacted in a solvent capable of partially dissolving at least one, for example methanol, dimethylformamide, ethanol, dimethylacetamide, dimethylsulfoxide or isopropanol, to prepare a compound represented by the following formula (V):

[Formula Ⅴ]

Wherein: B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above.

The compound represented by Formula V reacts with an electrophile (preferably N-protected glycinal, more preferably Nt-butoxycarbonylglycine or bis-Nt-butoxycarbonylglycine) Which is low temperature (preferably under basic conditions with a non-nucleophilic base (preferably lithium bis-trimethylsilylamide, potassium bis-trimethylsilylamide, sodium bis-trimethylsilylamide or lithium diisopropylamide) −90 to 25 ° C.), and reacting under suitable solvents (preferably tetrahydrofuran, diethyl ether or dioxane) which can dissolve at least one of the reactants to form a compound of formula VI:

[Formula VI]

Wherein: B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above; R ₄ and R ₅ each independently represent a hydrogen atom or a nitrogen-containing protecting group that can be removed quickly. Preferred are the R ₄ and R ₅ hydrogen atoms, t-butoxycarbonyl, benzyloxycarbonyl or benzyl.

The compound represented by the formula (IV) is preferably reduced by hydrogen gas under a suitable metal catalyst to prepare a compound represented by the following formula (X);

[Formula Ⅶ]

Wherein: B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above; R ₄ and R ₅ are the same as defined in Chemical Formula VI.

The compound represented by the formula (VIII) may be prepared in the presence of a non-nucleophilic base (preferably triethylamine or diisopropylethylamine) and an acylating agent or sulfonylating agent in a solvent capable of at least partially dissolving the reactants. Preferably with methinesulfonyl chloride or p-toluenesulfonyl chloride) to activate the hydroxy group. The activated hydroxy group is substituted with a suitable nucleophilic material (preferably thio acid salt, more preferably potassium thioacetate) to produce a compound represented by the formula

[Formula Ⅷ]

Wherein: A, B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above; R ₄ and R ₅ are as defined in formula VI, Ac represents an acyl group, preferably an acetyl group.

In addition, the compound represented by the formula (VIII) is chemically treated with triphenylphosphine, diethyl azadicarboxylate or dimethyl azadicarboxylate, and an acidic nucleophilic substance (preferably thioacetic acid) in a suitable solvent. It can also be converted into a compound represented by the above formula (VII).

The compound represented by the formula (V) is a nucleophilic base (preferably potassium carbonate, in the presence of an alcoholic solvent (preferably methanol, ethanol or isopropanol), an alkylating agent (preferably dimethyl chloromalonate, diethyl chloromalonate) Sodium carbonate, sodium hydroxide or potassium hydroxide) to prepare a compound of formula

[Formula Ⅸ]

Wherein: A, B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above; R ₄ and R ₅ are the same as defined in Chemical Formula VI.

The compound represented by the formula (VII) removes the protecting groups R ₄ and R ₅ simultaneously or under one of the appropriate conditions to prepare a compound represented by the following formula (VII):

[Formula Ⅹ]

Wherein: A, B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above.

When t-butoxycarbonyl (t-BOC) is used as the protecting group, a compound represented by the above formula (VII) is prepared by appropriate treatment with trifluoroacetic acid and then neutralization.

The compound represented by the formula (VII) is reacted with an alkylating agent (preferably trimethyloxonium tetrafluoroborate or triethyloxonium tetrafluoroborate) in a suitable solvent (preferably dichloromethane) to give lactim ether. Manufacture. The lactim ether intermediate compound is reacted with guanidine in an alcoholic solvent (preferably methanol, ethanol or isopropanol) to prepare a compound of formula XI:

Formula XI

Wherein: A, B, C and Y are each as defined in formula (I) above; R ₆ is as defined in Formula II above.

In addition, the compound represented by the formula (VII) is a thiolating agent (preferably P ₂ S ₅ or 2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphane-2 , 4-disulfide) can be converted to the compound represented by the above-mentioned formula (XI) which is a thiolactam intermediate. The intermediate compound is then alkylated using an alkylating agent (preferably methane iodide, trimethyloxonium tetrafluoroborate or triethyloxonium tetrafluoroborate), and an alcoholic solvent (preferably methanol, ethanol or isopropanol) To react with guanidine to prepare a compound represented by Formula XI.

The compound represented by Chemical Formula XI is hydrolyzed under basic conditions to prepare a compound represented by the following Chemical Formula XII:

[Formula XII]

In the formula: A, B, C and Y are as defined in formula (I), respectively.

In the case of the compound represented by Formula XI, wherein R ₆ is a hydrogen atom, the hydrolysis reaction may be omitted, and the compound represented by Formula XI is peptide-bonded as described below.

The compound represented by Formula XI (or the compound represented by Formula XI wherein R ₆ is a hydrogen atom) is present in free carboxylic acid form and forms a peptide bond with glutamic acid diester hydrogen chloride by a well-known method. Obtain the compound shown:

[Formula XIII]

Wherein: A, B, C and Y are each as defined in formula (I) above; R ₁ and R ₂ each independently represent, for example, C ₁ to C ₆ alkyl, hydroxyalkyl, alkylaryl or arylalkyl as ester groups which are bonded to CO ₂ and rapidly hydrolyzed.

Finally, when preparing a compound in free acid form, the compound represented by Chemical Formula XIII is hydrolyzed to obtain a compound represented by Chemical Formula I wherein R ₁ and R ₂ are each hydrogen atoms.

For compounds represented by formula (I) in which X is not CH ₂ , it may be prepared using an olefin represented by formula (XIV):

[Formula XIV]

Wherein: B, C and Y are each as defined in formula (I) above; X is as defined in the compound represented by the formula (I) except for CH ₂ ; R ₆ is as defined in Formula II above.

When X is a sulfur atom, an oxygen atom or a substituted or unsubstituted amino group, the compound represented by the formula (XIV) is prepared by alkylating the compound represented by the following formula (XV):

[Formula XV]

Wherein: B, C and Y are each as defined in formula (I) above; X represents a sulfur atom, an oxygen atom or a substituted or unsubstituted amino group; R ₆ is as defined in Formula II above.

The alkylation reaction is reacted with an alkyl halide (preferably allyl bromide) in the presence of a non-nucleophilic base (preferably triethylamine or diisopropylethylamine) to produce the compound represented by formula (XIV).

Wherein X is a substituted or unsubstituted C ₁ to C ₂ alkyl group except CH ₂ , a substituted or unsubstituted C ₂ to C ₃ alkenyl group, or a substituted or unsubstituted C ₂ to C ₃ alkynyl group, The compound is prepared by olefination with an aldehyde compound represented by the following formula (XVI):

[Formula XVI]

Wherein: B, C and Y are each as defined in formula (I) above; Z represents a substituted or unsubstituted C ₁ -C ₂ alkyl group except for CH ₂ , a substituted or unsubstituted C ₂ -C ₃ alkenyl group, or a substituted or unsubstituted C ₂ -C ₃ alkynyl group.

Aldehyde compounds represented by the above formula (XVI) are analogous methods [Chuan Shih et al., Journal of Medicinal Chemistry , vol. 35 (1992), 1109-116. Olefinization of the aldehyde is possible by using a methylene transition reagent (preferably methylene triphenylphosphoran).

The compound represented by the formula (XIV) is reacted with a dihydroxylating agent (preferably osmium tetroxide) in the presence of a suitable oxidizing agent (preferably N-methylmorpholine-N-oxide) to obtain a compound represented by the following formula (XVII) Manufactures:

Formula XVII]

Wherein: B, C and Y are each as defined in formula (I) above; X is as defined in formula (I) except for CH ₂ ; R ₆ is as defined in Formula II above.

The compound represented by the formula (XVII) is reacted with a sulfonylating agent (preferably methanesulfonyl chloride or p-toluenesulfonyl chloride) in the presence of a non-nucleophilic base (preferably triethylamine or diisopropylethylamine). Prepare mono-sulfonated intermediate compounds. This intermediate compound is reacted with a strong base (preferably sodium hydride) to produce a compound represented by the following formula (XVIII):

[Formula XVIII]

In the formula: B, C, Y and R ₆ are as defined in formula (XVII).

The epoxy compound represented by the formula (XVIII) is reacted with a nitrogenous nucleophilic material (preferably sodium azide) under a mild Lewis acid catalyst (preferably lithium perchlorate or magnesium perchlorate) to prepare an alcohol azide intermediate compound. This intermediate compound is reduced with hydrogen gas in the presence of a metal catalyst and subsequently protected with a suitable nitrogen-protecting group (preferably t-butoxycarbonyl, benzyloxycarbonyl or benzyl), represented by the following formula (VII ') Prepare the compound:

Formula VII ']

Wherein: B, C, X, Y and R ₆ are as defined for Formula XVII above; R ₄ and R 5 are each as defined in Formula VI above.

The compound represented by the above formula (VII ') is an acylating agent or sulfonylating agent in the presence of a non-nucleophilic base (preferably triethylamine or diisopropylethylamine) and in a solvent capable of partially dissolving at least one of the reactants. (Preferably methanesulfonyl chloride, p-toluenesulfonyl chloride) to activate the hydroxy group. The activated hydroxy group is substituted with a suitable nucleophilic material (preferably thio acid salt, more preferably potassium thioacetate) to produce a compound represented by the following formula (VIII '):

Formula VIII '

Wherein: A is as defined in formula (I) above; B, C, X, Y and R ₆ are as defined for Formula XVII above; R ₄ and R ₅ are each as defined in Formula VI above; And Ac represents an acyl group, preferably an acetyl group.

In addition, the compound represented by the above formula (VII ') may be chemically prepared using triphenylphosphine, diethyl azadicarboxylate or dimethyl azadicarboxylate, and an acidic nucleophilic substance (preferably thioacetic acid) in a suitable solvent. It can also be converted into the compound represented by the general formula (VIII ') by treatment.

The compound represented by the formula (VIII ') is a nucleophilic base (preferably potassium carbonate) in the presence of an alcoholic solvent (preferably methanol, ethanol or isopropanol), an alkylating agent (preferably dimethyl chloromalonate, diethyl chloromalonate) , Sodium carbonate, sodium hydroxide or potassium hydroxide) to prepare a compound represented by the following formula (IX '):

[Formula IX ']

Wherein: A is as defined in formula (I) above; B, C, X, Y and R ₆ are as defined for Formula XVII above; R ₄ and R ₅ are each as defined in Formula VI above.

The compound represented by the formula (IX ') is prepared by removing the protecting groups R ₄ and R ₅ at the same time or under one of the appropriate conditions under the appropriate conditions to prepare the compound represented by the following formula (X'):

Formula X '

Wherein: A is as defined in formula (I) above; B, C, X, Y and R ₆ are as defined in formula (XVII) above.

When t-butoxycarbonyl is used as a protecting group, the protecting group is treated with trifluoroacetic acid and then neutralized to prepare a compound represented by the above formula (X ').

The compound represented by the formula (X ') is reacted with an alkylating agent (preferably trimethyloxonium tetrafluoroborate or triethyloxonium tetrafluoroborate) in a suitable solvent (preferably dichloromethane) to lactim ether To prepare. The lactim ether intermediate compound is reacted with guanidine in an alcoholic solvent (preferably methanol, ethanol or isopropanol) to produce a compound of formula XI ':

Formula XI '

In the above formula; A is as defined in Formula I above: B, C, X, Y and R ₆ are as defined in Formula XVII above.

In addition, the compound represented by the formula (X ') is a thiolating agent (preferably P ₂ S ₅ or 2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphane- 2,4-disulfide) is used to react with the compound represented by the formula (X ') to convert to the compound represented by the formula (XI'), which is a thiolactam intermediate. The intermediate compound is then alkylated using an alkylating agent (preferably methane iodide, trimethyloxonium tetrafluoroborate or triethyloxonium tetrafluoroborate), and an alcoholic solvent (preferably methanol, ethanol or isopropanol) To react with guanidine to prepare a compound represented by Formula XI ′.

The compound represented by Formula XI 'is hydrolyzed in the presence of a base to prepare a compound represented by Formula XII':

Formula XII '

In the above formula; A is as defined in Formula I above, and B, C, X and Y are as defined in Formula XVII above.

In the case of the compound represented by the formula (XI ') wherein R ₆ is a hydrogen atom, the hydrolysis process is not an essential process, and the compound represented by the formula (XI') forms a peptide bond as shown below.

The compound represented by Chemical Formula XII '(or a compound represented by Chemical Formula XI' wherein R ₆ is a hydrogen atom) is present in free carboxylic acid form and forms a peptide bond with glutamic acid diester hydrogen chloride by a well-known method. Obtain the compound represented by XIII ':

Formula XIII '

In the above formula; A is as defined in formula (I) above; B, C, X, Y and R ₆ are as defined for Formula XVII above; R ₁ and R ₂ each independently represent, for example, C ₁ to C ₆ alkyl, hydroxyalkyl, alkylaryl or arylalkyl as ester groups which are bonded to CO ₂ and rapidly hydrolyzed.

Finally, when preparing a compound in free acid form, the compound represented by Chemical Formula XIII ′ is hydrolyzed to obtain a compound represented by Chemical Formula I wherein R ₁ and R ₂ are each hydrogen atoms.

Examples of preferred compounds among the compounds represented by the above formula (I) are as follows:

(4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl) -Ethyl] -2,5-thienoylamino-L-glutamic acid) diethyl ester;

4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Ethyl] -2,5-thienoylamino-L-glutamic acid;

4- [3- (2-Amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Propyl] -2,5-thienoylamino-L-glutamic acid;

4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Ethyl] -3-methyl-2,5-thienoylamino-L-glutamic acid; And

4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Ethyl] -2,5-furanoylamino-L-glutamic acid.

The compound represented by Formula (I) has one or more chiral centers. The present invention includes racemic mixtures and diastereomeric mixtures, in which the optically active compounds are separated by conventional methods.

The preparation of the compound represented by the formula (I) of the present invention will be described in more detail based on the following examples.

EXAMPLE

Example 1

4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Ethyl] -2,5-thienoylamino-L-glutamic acid, (compound 1)

Preparation of Methyl-5-bromo-4-methylthiophenecarboxylate 2:

Compound 2 (CAS Reg. No. [62224-19-5]] was prepared by S. Gronowitz, Ark. Kemi 8, 1955, 87, and SO Lawesson, Ark. Kemi 11, 1957, 337 . .

Preparation of 5-ethynyl-2-carbomethoxy thiophene 3:

Bromide compound 3 (5 g, 22.7 mmol), tris-triphenylphosphinepalladium (II) chloride (160 mg, 0.23 mmol) and copper iodide (65 mg, 0.34 mmol) were dissolved in diethylamine (75 mL), and then Trimethylsilyl acetylene (4.8 mL, 34 mmol) was added thereto. The reaction solution was stirred at 25 ° C. After 18 hours, the reaction solvent was removed under reduced pressure, and the residue was dissolved in diethyl ether (Et ₂ O; 300 mL), and extracted with 1N HCl (100 mL) and saturated bicarbonate solution (100 mL). The organic layer was dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography, and yellow solid silyl-protected compound (4.4 g, yield 81%) was obtained using 5% Et ₂ O / hexane as the eluting solvent. This compound was used directly in the next reaction.

The silyl-protected acetylene (4.4 g, 18.5 mmol) prepared above was dissolved in dimethylformamide (DMF; 50 mL), followed by addition of potassium fluoride monohydrate (3.3 g, 35.2 mmol). After stirring at 25 ° C. for 15 minutes, Et ₂ O (500 mL) was poured into the reaction mixture, and 100 mL of water was extracted four times (4 × 100 mL). The organic solution was dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography, and the target compound 3 (2.65 g, yield 86%) was obtained as an orange solid using 10% Et ₂ O / hexane as the eluting solvent.

NMR (CDCl ₃ ) δ: 3.45 (s, 1H), 3.88 (s, 3H), 7.22 (d, 1H, J = 3.5 Hz), 7.65 (d, 1H, J = 3.5 Hz).

Elemental Analysis for C ₈ H ₆ SO ₂ :

Theoretic value: C, 57.81; H, 3. 64; S, 19.29.

Found: C, 57.91; 3.71; S, 19.18.

The process of removing the silyl group is as follows.

Anhydrous K ₂ CO ₃ (3.7 g, 0.02 mmol) was suspended in methanol (700 mL), followed by addition of the silly-protected acetylene solid (59.7 g) prepared above. The suspension was stirred at 35 ° C. for 2 hours and then methanol was distilled off under reduced pressure. Water (50 mL) was added to the residue, followed by extraction with Et ₂ O (3 × 150 mL). The organic layer was dried over Na ₂ SO ₄ and distilled under reduced pressure. The residue was purified by silica gel fresh chromatography, and the target compound 3 (38.72 g, yield 98%) was obtained using 7% Et ₂ O / hexane as the eluting solvent.

Preparation of N- (t-butoxycarbonyl) -glycinal 4:

N- (t-butoxycarbonyl) -2-hydroxy-ethylamine (manufactured by Sigma Chemical; 2 g, 12.4 mmol) was dissolved in CH ₂ Cl ₂ (80 mL), followed by dimethyl sulfoxide ( DMSO: 1.3 mL, 18.6 mmol) and oxalyl chloride (1.2 mL, 13.7 mmol) were added. After 5 minutes, triethylamine (5.4 ml, 38.4 mmol) was added, and the temperature was raised to 25 ° C. Et ₂ O (200 mL) was poured into the reaction mixture and extracted with water (100 mL), 0.5N HCl (100 mL) and saturated bicarbonate solution (100 mL). The organic solution was dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure. The aldehyde residue was taken up in benzene (100 mL) and the solvent was removed under reduced pressure. This residue was an unstable compound, which was immediately used as an intermediate for the next reaction.

Preparation of 4-N- (t-butoxycarbonyl) -3-hydroxy-1- (2-carbomethoxy-5-thiophene) -butyn 5:

Acetylene compound 3 (1.7 g, 10.2 mmol) was dissolved in anhydrous tetrahydrofuran (THF; 10 mL), and then a solution containing 1M lithium bis-trimethylsilylamide (14.3 mL, 14.0 mmol) in THF was dissolved at -78 ° C. Was added. After 10 minutes, the aldehyde residue 4 prepared above was added dropwise to anhydrous THF (4 mL). The solution was stirred at −78 ° C. for 10 minutes and then heated to 0 ° C. over 20 minutes. Saturated ammonium chloride solution (5 mL) was added. Et ₂ O (200 mL) was poured into the reaction mixture, which was washed sequentially with water (100 mL) and saturated NaCl solution (100 mL). The organic solution was dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure. The residue was purified by silica gel flash chromatography, and the title compound (10.40% EtOAc / hexane) was used to obtain the desired alcohol compound 5 (0.96 g, 29% yield) as a light yellow oil.

NMR (CDCl ₃ ) δ: 1.46 (s, 9H), 3.38 and 3.58 (AB, 2H, J = 3, 7Hz), 3.88 (s, 3H), 4.71 (dt, 1H, J = 2.9, 5.2Hz), 5.05 (brs, 1 H), 7.15 (d, 1 H, J = 3.9 Hz), 7.64 (d, 1H, J = 3.9 Hz).

IR (neat): 2978.3, 1729.5, 1685, 1523, 1452, 1368, 1286, 1167, 1098, 959, 822, 750.4 cm ^-1

High performance mass spectrum results for C ₁₅ H ₁₉ NO ₅ S, M ⁺ Cs ⁺ :

Calculation: 458.0038

Measure: 458.0051

Preparation of Methyl-5-[(4-N- (t-butoxycarbonyl) -amino-3-hydroxy) -butyl] -thienyl-2-carboxylate 6:

The acetylene compound 5 (940 mg, 2.89 mmol) prepared above was dissolved in ethyl acetate (EtOAc, 50 mL), and 5% Pd / C (320 mg) was added thereto. The reaction mixture was stirred at 40 psi hydrogen at 25 ° C. for 17 hours. The reaction mixture was filtered through celite (diatomaceous earth) and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography, and the target alcohol compound (800 mg, yield 84%) was obtained as yellow oil using 20% EtOAc / CH ₂ Cl ₂ as the eluting solvent.

NMR (CDCl ₃ ) δ: 1.44 (s, 9H), 1.81 (m, 2H), 2.28 (brs, 1H), 2.99 (m, 2H), 3.10 and 3.29 (AB, 2H, J = 3, 6.9Hz) , 3.74 (m, 1H), 3.86 (s, 3H), 4.89 (brs, 1H), 6.82 (d, 1H, J = 3.7 Hz), 7.63 (d, 1H, J = 3.7 Hz)

Elemental Analysis for C ₁₅ H ₂₃ NO ₅ S:

Theoretic value: C, 54.69; H, 7.04: N, 4.25; S, 9.73.

Found: C, 54 79; H, 7.02; N, 4.29; S, 9.63.

In addition, alcohol compound 6 was prepared by the following preparation method using bis-t-butoxycarbonyl allylamine 14.

Preparation of bis-t-butoxycarbonyl allylamine 14:

Allylamine (57.10 g, 1.0 mmol) and dimethylaminopyridine (DMAP; 1.2 g, 0.01 mmol) were dissolved in acetonitrile (500 mL), followed by (t-BOC) ₂ O (220 g, 1 mol) to acetonitrile ( 100 ml) was added and stirred for 6 hours. The reaction mixture was diluted with toluene (100 mL) and the solvent was distilled under reduced pressure at 60 ° C. As a result, the oil was dissolved in acetonitrile (400 mL) again, DMAP (1.2 g, 0.01 mol) was added, and then (t-BOC) ₂ O (220 g, 1 mol) was added to acetonitrile (100 mL). The dissolved solution was added slowly. The reaction mixture was stirred at 60 ° C. for 12 hours. NaHCO ₃ (100 mL) was added to the reaction solution, and the mixture was extracted with CH ₂ Cl ₂ (3 × 150 mL). The organic layer was washed with brine, dried over Na ₂ SO ₄ , and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography, and the title compound 14 (156.9 g, 63% yield) of a clear crystalline solid was obtained using 5-20% EtOAc / hexane as an eluting solvent.

Melting Point: 43 ~ 44 ℃

¹ H NMR (CDCl ₃ ) δ (ppm): 1.5 (s, 18H), 4.18 (dd, 2H, J = 15Hz, J = 1Hz), 5.14 (ddd, 2H, J = 15Hz, J = 10Hz, J = 1 Hz), 5.85 (ddt, 2H, J = 10 Hz, J = 5 Hz, J = 1 Hz).

IR (KBr): 2978, 2935, 2860, 1724, 1689, 1342, 1130 cm ^-1 .

Elemental Analysis for C ₁₃ H ₂₃ NO ₄ :

Theoretic value: C, 60.68; H, 9.0 1; N, 5.44.

Found: C, 60 78; H, 9.0 4; N, 5.50.

Preparation of 1- (bis-t-butoxycarbonyl amino) -2-ethanal 15:

Bis-t-butoxycarbonyl allylamine 14 (0.60 g, 2.34 mmol) was dissolved in CH ₂ Cl ₂ (20 mL), and then ionized (40 volts, 500 amps, 1.0 until blue light at -78 ° C). L / min.O ₂ @ 3 psi) and dimethyl sulfoxide (10 mL) was added. The reaction mixture was diluted with brine and extracted with CH ₂ Cl ₂ (3 × 50 mL). The organic layer was dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography to give the title compound 15 (603 mg, 99% yield) as a clear crystalline solid.

Melting Point: 37 ~ 39 ℃

¹ H NMR (CDCl ₃ ) δ (ppm): 1.50 (s, 18H), 4.38 (s, 2H), 9.55 (s, 1H).

IR (KBr): 2984, 2935, 2724, 1792, 1734, 1699, 1362, 1153 cm ^-1 .

Elemental Analysis for C ₁₂ H ₂₁ NO ₅ :

Theoretic value: C, 55.58; H, 8, 16; N, 5.40.

Found: C, 55.20; H, 8. 19; N, 5.19.

Preparation of Methyl-5-[(4-N- (t-butoxycarbonyl) -amino-3-hydroxy) -butyl] -thienyl-2-carboxylate 6:

Dissolve 5-ethynyl-2-carbomethoxy thiophene 3 (9.64 g, 59 mmol) in THF (250 mL), then add lithium hexamethyl disilazide (LiHMDS; 0.9M, 65.6 mL, 60 mmol). It was.

The reaction mixture was stirred at -78 ° C for 2 hours. 1- (bis-t-butoxycarbonyl amino) -2-ethanal 15 (15.6 g, 60 mmol) was dissolved in THF (40 mL) and then injected into the reaction solution using a syringe (cannula). The reaction mixture was stirred at −78 ° C. for 8 hours, quenched by addition of acetic acid solution (3.4 mL, 60 mmol) dissolved in methanol (10 mL), and the reaction mixture was heated up to 10 ° C. for 10 minutes. Then water (60 mL) was added. Extract with EtOAc (3 × 100 mL), organic extract layer was washed with dilute NaHCO ₃ , dried over Na ₂ SO ₄ and solvent removed under reduced pressure until the solution volume was approximately 50 mL. EtOAc (125 mL) was added to the residue, which was charged to a Parr bottle containing 5% Pd / C containing 7.38 g of 30% acetylene starting material, followed by hydrogenation for 24 hours under a flow of 55 psi H ₂ . . The hydrogenated residue was filtered through celite and the filtrate was washed with a mixture of mitanol (100 mL) and EtOAc (100 mL).

The solvent was distilled under reduced pressure, and the residue was purified by silica gel fresh chromatography, and 50% EtOAc / hexane was used as the eluting solvent, and the solvent was distilled under reduced pressure. The residue is dissolved in methanol (30 mL) azeotropic with benzene and redissolved with anhydrous methanol (20 mL). The mixture was poured into 2M prop methoxide / methanol (60 mL), stirred for 45 minutes, then diluted HCl (0.1 M, 50 mL) was added and extracted with EtOAc (3 × 75 mL). The organic layer was washed with pH7 buffer (1M), dried over Na ₂ SO ₄ and distilled under reduced pressure. The residue was purified by silica gel fresh chromatography, and the target compound 6 (9.32 g, yield 48%) was obtained using 40% EtOAc / hexane as an eluting solvent.

Preparation of Methyl-5-[(4-N- (t-butoxycarbonyl) -amino-3-acetylthio) -butyl] -2-thiophenecarboxylate 7:

Alcohol compound 6 (9.26 g, 28.1 mmol) was dissolved in THF (100 mL), and triethylamine (TEA; 5.9 mL, 42 mmol) and methanesulfonyl chloride (2.4 mL, 31 mmol) were added. After 20 minutes, DMF (100 mL) and potassium thioacetate (12.8 g, 110 mmol) were added, and the temperature was raised to 25 ° C. After 3 days water (500 mL) was poured into the reaction mixture and extracted with Et ₂ O (800 mL). The organic layer was washed with water (200 mL), 1N HCl (200 mL), saturated bicarbonate solution (200 mL) and saturated NaCl solution (100 mL). The organic solution was dried over MgSO ₄ and the solvent was removed under reduced pressure. The residue was purified by silica gel flash chromatography, and the title compound was used as 25% EtOAc / hexane to afford thioacetate compound 7 (10.3 g, yield 95%) as a yellow oil.

NMR (CDCl ₃ ) δ: 1.44 (s, 9H), 1.91 and 2.04 (ABm, 2H), 2.37 (s, 3H), 2.96 (m, 2H), 3.36 (m, 2H), 3.61 (m, 1H) , 3.86 (s, 3H), 4.74 (brs, 1H), 6.79 (d, 1H, J = 3.6 Hz), 7.62 (d, 1H, J = 3.6 Hz).

IR (neat): 3366, 2976.4, 1713, 1520, 1462.1, 1366, 1290.5, 1267.3, 1169, 1098, 752, 631 cm ^-1

High performance mass spectrum analysis for C ₁₇ H ₂₅ NO ₅ S ₂ , M ⁺ Cs ⁺ :

Theoretic value: 520.0229.

Found: 520.0240.

Preparation of Methyl-5-[(4-N- (t-butoxycarbonyl) -amino-3- (dimethylmalonyl) thio) -butyl] -2-thiophenecarboxylate 8:

The thioacetate 7 prepared above (10.2 g, 26 mmol) was dissolved in anhydrous methanol, and then K ₂ CO ₃ (7.2 g, 52 mmol) and dimethyl chloromalonate (3.7 mL, 29 mmol) were added at 0 ° C. . After 3 hours, water (500 mL) was poured into the reaction mixture and extracted with Et ₂ O (3 × 500 mL). The organic layer was washed sequentially with water (500 mL) and saturated NaCl solution (100 mL), dried over MgSO ₄ , and the solvent was removed under reduced pressure. The residue was purified by silica gel flash chromatography, and the thioether compound 8 (11.46 g, 93% yield) was obtained as a light yellow oil using 30% EtOAc / hexane as an eluting solvent.

NMR (CDCl ₃ ) δ: 1.44 (s, 9H), 1.85 and 2.00 (ABm, 2H), 3.03 (m, 3H), 3.30 (m, 2H), 3.80 (s, 6H), 3.86 (s, 3H) , 5.10 (brs, 1 H), 6.82 (d, 1 H, J = 3.8 Hz), 7.63 (d, 1H, J = 3.8 Hz).

IR (neat): 3442, 2974, 2995, 1734, 1713, 1512, 1460, 1435, 1366, 1291, 1267, 1167, 1098, 1022, 752 cm ^-1

High performance mass spectrum analysis for C ₂₀ H ₂₉ NO ₈ S ₂ , M ⁺ Cs ⁺ :

Theoretical value: 608.0389.

Found: 608.0370.

Of 6-[(5-carbomethoxythien-2-yl) -ethyl] -2-carbomethoxy-3-oxo-3,4,5,6-tetrahydro- [1,4] -thiazine 9 Produce :

The thioether 8 (470 mg, 0.99 mmol) prepared above was dissolved in CH ₂ Cl ₂ and then trifluoroacetic acid (TFA; 4 mL) was added at 0 ° C. After 1.5 h, saturated bicarbonate solution (50 mL) was poured into the reaction mixture and extracted with CH ₂ Cl ₂ (4 × 100 mL). The solvent was removed under reduced pressure, and the residue was dissolved in methanol (10 mL). After stirring at 25 ° C. for 1.5 hours, the solvent was removed under reduced pressure and the residue was purified by silica gel flash chromatography, and the lactam compound as a colorless oil using 40% EtOAc / CH ₂ Cl ₂ as the eluting solvent. 9 (298 mg, yield 88%) was obtained.

NHR (CDCl ₃ ) δ: 1.94 (m, 2H), 3.01 (m, 2H), 3.4 to 3.7 (m, 4H), 3.80 and 3.82 (s, 3H), 3.87 (s, 3H), 6.25 (brs, 1H), 6.83 (d, 1H, J = 3.7 Hz), 7.64 (d, 1H, J = 3.7 Hz).

IR (KBr): 2951, 1732, 1669, 1462, 1294, 1267, 1194, 1157, 1098, 1005, 752 cm ^-1

Elemental Analysis for C ₁₄ H ₁₇ NO ₅ S ₂ :

Theoretic value: C, 48.96; H, 4.99; N, 4.08, S, 18.67.

Found: C, 49.06; H, 4.93; N, 4.09; S, 18.60.

Methyl- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] thiazin-6-yl) -ethyl ] -2,5-Tenoate 10 Preparation:

The lactam 9 (1.9 g, 5.5 mmol) prepared above was dissolved in anhydrous CH ₂ Cl ₂ (150 mL), and trimethyloxonium tetrafluoroborate (1.06 g, 7.2 mmol) was added all at once. The reaction solution was stirred at 25 ° C. for 6 hours, then added with 10% K ₂ CO ₃ aqueous solution (50 mL), and extracted with CH ₂ Cl ₂ (3 × 200 mL). The organic layer was dried over MgSO ₄ and the solvent was removed under reduced pressure. And immediately used for the next reaction.

Anhydrous guanidine hydrogen chloride (1.58 g, 16.6 mmol) was added to the fractionation flask, and anhydrous methanol (600 mL) was added thereto. After passing anhydrous argon gas in the solution for 10 minutes, anhydrous sodium methoxide (926 mg, 17.1 mmol) was added. This mixture was added to a solution in which the lactam ether prepared above was dissolved in methanol (20 mL) using a cannula. It was refluxed for 20 hours under an argon stream. Adjust to pH 4 with 1N HCI and remove solvent under reduced pressure. The residue was taken up in CHCl ₃ (500 mL) and washed with water (2 × 100 mL). The organic layer was dried over MgSO ₄ and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography, and the target compound (413 mg) was obtained as a white solid using 5-10% methanol / CH ₂ Cl ₂ as the eluting solvent. It was then dissolved in hot methanol (25 mL) and cooled slowly to 4 ° C. for 18 h. The desired pyrimidinone 10 (180 mg, yield 9%) produced as a light yellow solid was collected by filtration.

NMR (d ₆ -DMSO) δ: 1.72 (m, 1H), 1.88 (m, 1H), 2.8-3,22 (m, 4H), 3.50 (dt, 1H, J = 2.8 Hz), 5.99 (brs, 2H), 6.64 (brs, 1H), 6.98 (d, 1H, J = 3.8 Hz), 7.62 (d, 1H, J = 3.8 Hz), 10.02 (brs, 1H).

Elemental Analysis for C ₁₄ H ₁₆ N ₄ O ₃ S ₂ , M ⁺ Na ⁺ :

Theoretical value: 375.0562.

Found: 375.0550.

In addition, pyrimidinone 10 can also be manufactured by the following method.

Lactam 9 (500 mg, 1.46 mmol) was dissolved in anhydrous THF (20 mL) and then Lawesson's reagent (2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4- Diphosphetane-2,4-disulfide; 648 mg, 1.60 mmol) was added. Stir at 25 ° C. for 20 h, then add saturated bicarbonate solution (50 mL) and extract with EtOAc (2 × 200 mL).

The organic layer was washed with saturated NaCl solution (50 mL), dried over MgSO ₄ , and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography, and thiollactam 11 (525 mg) was obtained using 8% EtOAc / CH ₂ Cl ₂ as the eluting solvent.

The compound was immediately used for the next reaction.

Thiolactam 11 (525 mg) prepared above was dissolved in anhydrous THF (10 mL), and then 1N NaOH (1.6 mL) was added thereto. Iodomethane (0.1 mL, 1.6 mmol) was added thereto to obtain methylated thiolactam, which was used for the next reaction without purification. Anhydrous guanidine hydrogen chloride (1.39 g, 14.6 mmol) was added to the fractionation flask, and anhydrous methanol (300 mL) was added thereto. After passing anhydrous argon gas into the solution for 10 minutes, anhydrous sodium methoxide (796 mg, 14.7 mmol) was added. This mixture was added to the methylated thiollactam (20 mL) prepared above using a cannula. It was refluxed for 48 hours under argon stream. Adjust to pH 4 with 1N HCl and remove solvent under reduced pressure. The residue was taken up in CHCl ₃ (500 mL) and washed with water (2 × 100 mL). The organic layer was dried over MgSO ₄ and the solvent was removed under reduced pressure. The residue was purified by silica gel fresh chromatography, and 5 to 10% methanol / CH ₂ Cl ₂ was used as the eluting solvent. It was then dissolved in hot methanol (20 mL) and slowly cooled to 4 ° C. for 18 h. The target pyrimidinone 10 (131 mg, yield 9%) was collected by filtration.

[2- (2-Amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] thiazin-6-yl) -ethyl]- Preparation of 2,5-thienoic acid 12:

Pyrimidinone 10 (180 mg, 0,51 mmol) was added to a 1N HaOH aqueous solution (5 mL) and stirred at 25 ° C for 20 hours. After cooling to 0 ° C., the pH of the solution was adjusted to 2 with 1N HCl. The brown solid was filtered, washed with water and dried under reduced pressure to give the desired light brown acid compound 12 (111 mg, yield 64%).

NMR (d ₆ -DMSO) δ: 1.72 (m, 1H), 1.92 (m, 1H), 2.78-3.68 (m), 6.07 (brs, 2H), 6.68 (brs, 1H), 6.92 (d, 1H, J = 3.7 Hz), 7.52 (d, 1H, J = 3.7 Hz), 10.12 (brs, 1H), 12.89 (brs, 1H).

High performance mass spectrum analysis for C ₁₃ H ₁₄ N ₄ O ₃ S ₂ , M ⁺ :

Theoretic value: 338.0507.

Found: 338.0517.

4- [2- (2-Amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] thiazin-6-yl) -ethyl ] -2,5-thienoylamino-L-glutamic acid diethyl ester 13

The acid compound 12 (110 mg, 0.33 mmol) prepared above was dissolved in anhydrous DMF (5 mL), and then 1-hydroxy-benzotriazole hydrate (48 mg, 0.36 mmol) and diisopropylethylamine (62 μl) were added thereto. , 0.36 mmol), glutamic acid diethyl ester hydrate (86 mg, 0.36 mmol) and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide methoxide (106 mg, 0.36 mmol) were added. The reaction solution was stirred at 25 ° C. under argon for 20 hours, poured water (50 mL) into the reaction mixture, and extracted with EtOAc (2 × 150 mL). The organic layer was extracted with water (3 × 50 mL) and saturated NaCl solution, dried over MgSO ₄ , and the solvent was removed under reduced pressure. The residue was purified by silica gel flash chromatography, and the title compound 13 (120 mg, yield 71%) was obtained as a light yellow amorphous solid using 0-10% methanol / CH ₂ Cl ₂ as the elution solvent.

NMR (d ₆ -Acetone) δ: 1.20 (m, 6H), 2.2 (m, 1H), 2.46 (t, 1H, J = 7.6 Hz), 2.78 (s, 1H), 2.82 (s, 1H), 2.92 -3.1 (m, 2H), 3.41 (m, 1H), 3.72 (dt, 1H, J = 3.0, 12.7 Hz), 4.02-4.2 (m, 4H), 4.58 (m, 1H), 5.90 (brs, 1H ), 6.05 (brs, 1H), 6.90 (d, 1H, J = 3.8 Hz), 7.58 (d, 1H, J = 3.8 Hz), 7.78 (d, 1H, J = 8 Hz).

High performance mass spectrum analysis for C ₂₂ H ₂₉ N ₅ O ₆ O ₂ , M ⁺ H ⁺ :

Theoretic Value: 524.1638.

Found: 524.1650.

4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Ethyl] -2,5-thienoylamino-L-glutamic acid, Preparation of (Compound 1):

Diethyl ester 13 (115 mg, 0.22 mmol) prepared above was dissolved in 1N aqueous NaOH solution (3 mL), and then stirred at 25 ° C. for 14 hours. After cooling to 0 ° C., the pH of the reaction solution was adjusted to 3.5 using an aqueous hydrochloric acid solution. The resulting solid was filtered, washed with water and dried under reduced pressure to give the desired acid compound (82 mg, yield 80%) as a white solid.

NMR (d ₆ -DMSO) δ: 1.62 to 2.02 (m, 4H), 2.27 (m, 2H), 2.78 to 3.00 (m, 3H), 4.27 (dd, 1H, J = 6, 6.8 Hz), 6.00 ( brs, 2H), 6.63 (brs, 1H), 6.88 (d, 1H, J = 3.7 Hz), 7.63 (d, 1H, J = 3.7 Hz), 8.37 (d, 1H, J = 7.4 Hz), 10.05 ( brs, 1 H), 12.85 (brs, 2H).

IR (KBr): 3371, 1700, 1643, 1543, 1345 cm ^-1

High performance mass spectrum analysis for C ₁₈ H ₂₁ N ₅ O ₆ S ₂ , M ⁺ H ⁺ :

Theoretic value: 468.1012.

Found: 468.1025.

Elemental analysis for C ₁₈ H ₂₁ N ₅ O ₆ S ₂ .1.5H ₂ O:

Theoretic value: C, 43.71; H, 4.89; N, 14. 16; S, 12.97.

Found: C, 43.50; H, 4.67; N, 14.07; S, 12.72.

Biological and Biochemical Assessment

Determination of Inhibitory Activity of GAR Transferrillase:

GAR transsomerase (GARFT) analysis was measured by the following method using an analysis method by Young et al. [ Biochemistry 23, 3979-3986 (1984)]. The reaction mixture was catalyzed by human GARFT, 0-250 mM test compound, 20 μΜ glycineamide ribonucleotide (GAR), 10 or 20 μΜ N ¹⁰ -formyl-5,8-dydeazafolate (FDDF), 50 mM HEPES- KOH (pH 7.5) and 50 mM KCl. The reaction was initiated by enzyme addition and monitored for an increase in absorbance at a wavelength of 294 nm at 20 ° C. with addition to 11 nM concentration (e ₂₉₄ = 18.9 mM ⁻¹ Cm ⁻¹ ).

The GARFT inhibition constant (K _i ) depends on the extent to which the catalyst rate at steady state depends on the inhibitor and substrate concentration. Then, the actual form of the observed inhibition is dependent on the FDDF concentration, which is expressed as K _{i, app} is expressed by a formula expressed as _{K i, app = K i +} (K i / K m) [FDDF]. The Michaelis constant (K _m ) for FDDF was determined independently by the catalyst rate, which depends on the concentration of FDDF. K _m and K _i values are obtained by plotting the nonlinear method in the Michaelis equation or the competitive inhibition microphoneless equation. The data obtained from the tight-binding inhibition are analyzed and shown in Morrison's tight-binding equation (Morrison, Biochem Biophys Acta 185 (1969), 269-286) to determine the K _i value.

Cell line:

The types of cell lines used and their sources are shown in Table 1 below. The growth conditions and required media of each cell line are summarized in Table 2 below. All cell lines were incubated at 37 ° C., 5% carbon dioxide conditions in a humidified-incubator.

Growth inhibition in in vitro test:

The mother liquor of the inhibitor was prepared in 10 mM aqueous sodium hydrogen carbonate solution and stored in 1 ml aliquots at -20 ° C during cell culture experiments. Cell growth inhibition was measured by modifying the method proposed by Mosmann ( J. Immunol, Methods 65 (1983), 55-63). In each cell line, cells in the mid-log phase were placed in fresh RPMI growth medium (Mediatech, Washington, DC) with dialysis placental serum (Hyclone Laboratories Inc., Logan, UT). cells / ml), then aliquot into columns 2 to 12 on 96-well microtiter plates. Column 1 was filled with 135 mL fresh medium without adding cells for use as a blank. The plates are then placed in an incubator at 37 ° C., 5% carbon dioxide conditions. After 1 hour to 4 hours, the plates were removed from the incubator and 10 mL of the test substance, secondary dilution 15 mL / well, was added in 12 to 14 columns. For the opposite experiment, hypoxanthine (HX, 1.75 mM), or AICA (1.75 mM) was included in all drug solutions (final concentration 175 mM). Four wells containing each concentration of test compound were made on each plate. 15 ml of medium without test compound was added to the wells of column 1 on the plate. The cells were then placed back into the incubator and incubated at steady state. On day 3 for L1210 and L1210 / CI920 cells, or on day 5 for CCRF-CEM cells, tissue culture medium (0.8 mg / ml MTT 50 ml; (4,5-dimethylthiazole- 2-yl) -2,5-diphenyl tetrazolium bromide; Sigma catalog no.M2128) was added to each well on all plates. After 4 hours, all plates were removed from the incubator and centrifuged at 1200 rpm for 7 minutes. Media was absorbed and removed and 150 ml of DMSO was added to each well on all plates. The plates were then mixed at low speed with a Vortex mixer for 1 hour at room temperature, dark. The metabolic degree of MTT was measured at 540 nm by spectroscopic method using a spectrometer (Molecular devices Vmax kinetic microplate reader). The drug concentration required to reduce cell growth as measured by MTT metabolism was calculated by summing the value just above the OD (minus blank) and the value just below the 50% control OD (minus blank).

In vivo anticancer activity test:

On initiation day, 2 mm 2 of 6C3HED lymphoma segments were implanted subcutaneously under the armpits of 6 C3H / He mice. On the first day of the test, the test compound was administered intraperitoneally with 40% Encapsin, which was administered daily for 9 days. Control animals were not administered test compounds. The percent inhibition is shown in Table 4, which was determined by comparing the mass of the control tumor (only diluent) with the animals that received the test compound for 11 days.

On initiation day, 2 mm2 of C3H / BA breast cancer segments were implanted subcutaneously under the armpits of 6 C3H / He mice. On the first day of the test, the test compound was administered intraperitoneally with 40% Encapsin, which was administered daily for 9 days. Control animals were not administered test compounds. The percent inhibition is shown in Table 5 below, which was determined by comparing the mass of the control tumor (only diluent) with the animals that received the test compound for 14 days.

The embodiments described above can be further diversified within the scope of the invention. Appropriate applications are possible to those skilled in the art, which are also included in the scope of the present invention.

Within the scope of possible chemistry, the chemical groups exemplified in the present invention may be substituted. In some cases, this possibility is evident by enumeration (eg, substituted or unsubstituted C ₁ -C ₃ alkyl groups).

When at least one R ₆ is exemplified in any formula, each R ₆ is independently selected within a given possibility.

Claims (45)

A compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof. [Formula I]

In the above formula; A represents an oxygen atom, a sulfur atom or Se; X is a substituted or unsubstituted C ₁ -C ₃ alkyl group, a substituted or unsubstituted C ₂ -C ₃ alkenyl group, a substituted or unsubstituted C ₂ -C ₃ alkynyl group, a substituted or unsubstituted amino group, an antigen or An oxygen atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; And R ₁ and R ₂ each independently represent a moiety that is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group. The compound or salt according to claim 1, wherein X is CH ₂ , CH ₂ CH ₃ , NH, oxygen atom, sulfur atom, CH (CH ₂ OH) or NCH ₃ . The compound or salt according to claim 1, wherein Y is a sulfur atom or an oxygen atom, B is a hydrogen atom, and C is a hydrogen atom or CH ₃ . The compound or salt according to claim 1, wherein A is a sulfur atom, Y is a sulfur atom, and R ₁ and R ₂ are each hydrogen atoms. The compound or salt according to claim 1, wherein X is CH ₂ . The compound or salt according to claim ₁ , wherein R ₁ and R ₂ are each a hydrogen atom, a C _{1 to} C ₆ alkyl group, a hydroxyalkyl group, an alkylaryl group and an arylalkyl group. The compound or salt according to claim 6, wherein R ₁ and R ₂ are each independently a hydrogen atom or a C ₁ -C ₂ alkyl group. The compound or salt according to claim 7, wherein A is a sulfur atom, Y is a sulfur atom, and X is CH ₂ . The compound or salt according to claim 8, wherein B is a hydrogen atom, C is a hydrogen atom, and R ₁ and R ₂ are each independently a hydrogen atom. The compound or salt according to claim 1, wherein the compound or salt (4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl) -Ethyl] -2,5-thienoylamino-L-glutamic acid) diethyl ester; 4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Ethyl] -2,5-thienoylamino-L-glutamic acid; 4- [3- (2-Amino-4-oxo-4,6.7, 8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl) -propyl] -2,5-thienoylamino-L-glutamic acid; 4- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl)- Ethyl] -3-methyl-2,5-thienoylamino-L-glutamic acid; And 4- [2- (2-amino-4-oxo-4,6.7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4] -thiazin-6-yl) -ethyl] Selected from -2,5-furanoylamino-L-glutamic acid. (i) a compound represented by formula (I) or a pharmaceutically acceptable salt thereof; And (ii) A pharmaceutical composition effective as an anti-malignant tumor agent, characterized in that it consists of a pharmaceutical carrier. [Formula I]

In the above formula; A represents an oxygen atom, a sulfur atom or Se; X is substituted or unsubstituted C ₁ -C ₃ alkyl group, substituted or unsubstituted C ₂ -C ₃ alkenyl group, substituted or unsubstituted C ₂ -C ₃ alkynyl group, substituted or unsubstituted amino group, sulfur atom or oxygen Represents an atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; And R ₁ and R ₂ each independently represent a moiety that is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group. In the pharmaceutical composition according to claim 11, wherein X is CH ₂ , CH ₂ CH ₃ , NH, oxygen atom, sulfur atom, CH (CH ₂ OH) or NCH ₃ . The pharmaceutical composition according to claim 11, wherein Y is a sulfur atom or an oxygen atom, B is a hydrogen atom, and C is a hydrogen atom or CH ₃ . The pharmaceutical composition according to claim 13, wherein A is a sulfur atom, Y is a sulfur atom, and X is CH ₂ . In the pharmaceutical composition according to claim 14, wherein C is a hydrogen atom, and R ₁ and R ₂ are each independently a hydrogen atom. A method of inhibiting the growth or proliferation of cells of microorganisms or higher organisms by administering an effective amount of a compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof to mammals or birds except humans. [Formula I]

Wherein: A represents an oxygen atom, a sulfur atom or Se; X is substituted or unsubstituted C ₁ -C ₃ alkyl group, substituted or unsubstituted C ₂ -C ₃ alkenyl group, substituted or unsubstituted C ₂ -C ₃ alkynyl group, substituted or unsubstituted amino group, sulfur atom or oxygen Represents an atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; And R ₁ and R ₂ each independently represent a moiety that is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group. The method according to claim 16, wherein X is CH ₂ , CH ₂ CH ₃ , NH, oxygen atom, sulfur atom, CH (CH ₂ OH) or NCH ₃ . The method according to claim 16, wherein Y is a sulfur atom or an oxygen atom, B is a hydrogen atom, and C is a hydrogen atom or CH ₃ . The method according to claim 18, wherein A is a sulfur atom, Y is a sulfur atom, and X is CH ₂ . 20. The method according to claim 19, wherein C is a hydrogen atom and R ₁ and R ₂ are each independently a hydrogen atom. The compound represented by the following formula (VI). [Formula VI]

In the formula: Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; R ₆ represents a moiety that is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group, and R ₄ and R ₅ each independently represent a group capable of rapidly removing a hydrogen atom or a nitrogen protecting group. The compound of claim 21, wherein Y is a sulfur atom; B is a hydrogen atom; C is a hydrogen atom; R ₆ is a hydrogen atom, a C ₁ -C ₆ alkyl group, a hydroxyalkyl group, an alkylaryl group or an arylalkyl group; And R ₄ and R ₅ are each independently a hydrogen atom, a t-butoxycarbonyl group, a benzyloxycarbonyl group or a benzyl group. The compound according to claim 21, wherein the compound is 4-N- (t-butoxycarbonyl) -3-hydroxy-1- (2-carbomethoxy-5-thiophene) -butyne. The compound represented by following formula (VII). [Formula VII]]

Wherein X 'is a substituted or unsubstituted C ₁ -C ₃ alkyl group, a substituted or unsubstituted C ₂ -C ₃ alkenyl group, a substituted or unsubstituted C ₂ -C ₃ alkynyl group, substituted or unsubstituted Amino group, sulfur atom or oxygen atom; A represents an oxygen atom, a sulfur atom or Se; Y represents an oxygen atom, a sulfur atom or NH, and B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; R ₆ represents a moiety which is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group; And R ₄ and R ₅ each independently represent a group capable of removing a hydrogen atom or a nitrogen protecting group. 25. A compound according to claim 24, wherein X 'is CH ₂ ; Y is a sulfur atom: B is a hydrogen atom; C is a hydrogen atom; R ₆ is a hydrogen atom, a C ₁ -C ₆ alkyl group, a hydroxyalkyl group, an alkylaryl group or an arylalkyl group; And R ₄ and R ₅ are each independently a hydrogen atom, a t-butoxycarbonyl group, a benzyloxycarbonyl group or a benzyl group. The compound according to claim 24, wherein the compound is methyl-5-[(4-N- (t-butoxycarbonyl) -amino-3-hydroxy) -butyl] -thienyl-2-carboxylate Will. The compound represented by following formula (VII). [Formula VII]]

Wherein X 'is a substituted or unsubstituted C ₁ -C ₃ alkyl group, a substituted or unsubstituted C ₂ -C ₃ alkenyl group, a substituted or unsubstituted C ₂ -C ₃ alkynyl group, substituted or unsubstituted Amino group, sulfur atom or oxygen atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; R ₆ represents a moiety which is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group; R ₄ and R ₅ each independently represent a group capable of removing a hydrogen atom or a nitrogen protecting group; And Ac represents an acyl group. 28. A compound according to claim 27, wherein X 'is CH ₂ ; Y is a sulfur atom; B is a hydrogen atom; R ₆ is a hydrogen atom, a C ₁ -C ₆ alkyl group, a hydroxyalkyl group, an alkylaryl group or an arylalkyl group; And R ₄ and R ₅ are each independently a hydrogen atom, a t-butoxycarbonyl group, a benzyloxycarbonyl group or a benzyl group. 28. The compound of claim 27, wherein the compound is methyl-5-[(4-N- (t-butoxycarbonyl) -amino-3-acetylthio) -butyl] -2-thiophenecarboxylate . The compound represented by following formula (VII). [Formula VII]]

Wherein: A is an oxygen atom, a sulfur atom or a selenium atom; X 'is a substituted or unsubstituted C ₁ -C ₃ alkyl group, a substituted or unsubstituted C ₂ -C ₃ alkenyl group, a substituted or unsubstituted C ₂ -C ₃ alkynyl group, a substituted or unsubstituted amino group, a sulfur atom or An oxygen atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; R ₆ represents a moiety which is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group; And R ₄ and R ₅ each independently represent a group capable of removing a hydrogen atom or a nitrogen protecting group. A compound according to claim 30, wherein A is a sulfur atom; X 'is CH ₂ ; Y is a sulfur atom; B is a hydrogen atom; C is a hydrogen atom; R ₆ is a hydrogen atom, a C ₁ -C ₆ alkyl group, a hydroxyalkyl group, an alkylaryl group, or an arylalkyl group; And R ₄ and R ₅ are each independently a hydrogen atom, a t-butoxycarbonyl group, a benzyloxycarbonyl group or a benzyl group. 31. A compound according to claim 30, wherein the compound is methyl-5-[(4-N- (t-butoxycarbonyl) -amino-3-dimethylmalonyl) thio) -butyl] -2-thiophencarboxyl It is rate. The compound represented by following formula (VII). [Formula VII]]

Wherein: A is an oxygen atom, a sulfur atom or a selenium atom; X 'is a substituted or unsubstituted C ₁ -C ₃ alkyl group, a substituted or unsubstituted C ₂ -C ₃ alkenyl group, a substituted or unsubstituted C ₂ -C ₃ alkynyl group, a substituted or unsubstituted amino group, a sulfur atom or An oxygen atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; And R ₆ represents a moiety which is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group. 34. A compound according to claim 33, wherein A is a sulfur atom; X 'is CH ₂ ; Y is a sulfur atom; B is a hydrogen atom; C is a hydrogen atom; And R ₆ is a hydrogen atom, a C ₁ -C ₆ alkyl group, a hydroxyalkyl group, an alkylaryl group or an arylalkyl group. 34. The compound of claim 33, wherein the sulfur compound is 6-[(5-carbomethoxythien-2-yl) -ethyl] -2-carbomethoxy-3-oxo-3,4,5,6- Tetrahydro- [1,4] -thiazine. The compound represented by the following formula (XI "). Formula XI "

Wherein: A is an oxygen atom, a sulfur atom or a selenium atom; X 'is optionally substituted C ₁ ~C ₃ alkyl group, a substituted or unsubstituted C ₂ ~ C ₃ alkenyl group, a substituted or unsubstituted C ₂ ~C ₃ alkynyl group, a substituted or unsubstituted amino group, a sulfur atom, or An oxygen atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom; C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C ₁ -C ₆ alkyl group; And R ₆ represents a moiety which is bonded to a hydrogen atom or CO ₂ to form a hydrolyzable ester group. 37. A compound according to claim 36, wherein A is a sulfur atom; X 'is CH ₂ ; Y is a sulfur atom; B is a hydrogen atom; C is a hydrogen atom; And R ₆ is a hydrogen atom, a C ₁ -C ₆ alkyl group, a hydroxyalkyl group, an alkylaryl group or an arylalkyl group. 37. The compound of claim 36, wherein the compound is methyl- [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1 , 4] thiazin-6-yl) -ethyl] -2,5-thienoate. A compound represented by the following formula (XII). Formula XII "

Wherein: A is an oxygen atom, a sulfur atom or a selenium atom: X 'is a substituted or unsubstituted C ₁ -C ₃ alkyl group, a substituted or unsubstituted C ₂ -C ₃ alkenyl group, a substituted or unsubstituted C ₂ A -C ₃ alkynyl group, a substituted or unsubstituted amino group, a sulfur atom or an oxygen atom; Y represents an oxygen atom, a sulfur atom or NH; B represents a hydrogen atom or a halogen atom, and C represents a hydrogen atom, a halogen atom or a substituted or unsubstituted C _{1 to} C ₆ alkyl group. 40. The compound of claim 39, wherein A is a sulfur atom; X 'is CH ₂ ; And Y is a sulfur atom. 40. The compound of claim 39, wherein the compound is [2- (2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimido [5,4-6] [1,4 ] Thiazin-6-yl) -ethyl] -2,5-thienoic acid. A compound represented by the following formula.

The compound according to claim 42 is optically active. The compound according to claim 43 is represented by the following formula.

The compound according to claim 43 is represented by the following formula.