FI93962B - 6,7-Dihdyro-6-mercapto-5H-pyrazolo[1,2a][1,2,4]triazolium intermediates, and a process for preparing them - Google Patents

Description

93962 6,7-Dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium intermediates and process for their preparation

By division separated from the application 906 464.

5

The present invention relates to intermediates useful in the preparation of (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] - [1,2,4] of formula (I)). triazolium-6-yl)] thio-6 - [(R) -1-hydroxyethyl] -1-methylcarbapenem-10 3-carboxylate in the preparation of TtPC ^ 15 o ^ cooe or a pharmacologically acceptable salt thereof.

After the antibiotic "thienamycin" was found in nature [U.S. Patent 3,959,357; J. Am. Chem. Soc., 100 313 (1987)], many studies have been conducted to develop carbapenem antibiotics. Extensive research has resulted in the development of imipenem (INN), an almost available antibacterial agent for clinical use.

Some time ago, Kumagai et al. proposed (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6 - [(R ) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate as a carbapenem-30 antibiotic, see U.S. Patents 4,866,171, 4,925,836 and 4,925,935 and FI Patent 86,729. a penem-3-carboxylic acid compound in which the β-methyl group is attached to the 1-position and the 6,7-dihydro-5H-pyrazolo [1,2-a] - [1,2,4] triazolium-6-ylthio group described in follow the formula-35

-C/O

© 2 93962 is joined to the 2-position in the carbapenem backbone. Due to these particular substitutions, the compound of formula (I) has a better antibacterial activity and is chemically and physicochemically very stable in the living body and is very stable against dehydropeptidase (DHP) known as a renal enzyme.

To date, the only known synthesis of (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] - [1,2,4] -triazolium-6 of formula (I) -yl)] thio-6 - [(R) -1-hydroxyethyl] -10 1-methylcarbapenem-3-carboxylate

In Kumagai et al., Occurs from an imino intermediate. According to Eli Kumagai et al., A compound of formula (I) can be prepared by reacting a compound of formula (II): 15 ° H (n CH)

"/-OF

Wherein R 1 is a carboxyl protecting group and R n is an acyl group, to react with a mercaptor reagent of formula (VII) - N-R b - HS - (I (VII) 1 - N-R b wherein R b is an amino protecting group to give a compound of formula (VIII)). ):

Hr HH £ Hs / -N * Rb Ξ I - ((VIII) CHa H 1 T N-Rb (f — N- ^ U COOR1 35 3 93962 wherein R 1 and R b have the same meanings as above, and by performing removal of the protecting groups, R1 and Rb, for a compound of formula (VIII), a carbapenem compound of formula (IX) is obtained.

H * H H SHa A-NH

COOH

And then, by reacting the resulting compound of formula (IX) with a formimid acid ester, a carbapenem compound of formula (I) is obtained.

One possible reaction route for the formation of a compound of formula (I) by a reaction in which a compound of formula (IX) is reacted with a formimidic acid ester derivative is a reaction in which a compound of formula (IX) is reacted with a formimidic acid ester derivative to give the following intermediate: (1R, 5S, 6S) -6 - [(R) -1-hydroxyethyl] -1-methyl-2 - [(1,2-diiminomethyl) -4-pyrazolidinyl] thiocarbapenem-3- carboxylic acid: HO „CH_

- H H f 3 / -N-CH = NH

25 CH j

^ 3 H I || Y-CH = N-NH

J-N-k.

0 COOH

and this imino intermediate is an instantaneous compound and is subjected to a cyclization reaction to form a compound of formula (I).

The intermediate of the present invention relates to a study aimed at finding a process for the preparation of a compound of formula (I), (1R, 5S, 6S) -2 - [(6,7-dihydro-35-5H-pyrazolo [1,2- a] [1,2,4] triazolium-6-yl)] thio-6 - [(R) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate by a method other than the same as a process for the preparation of an imino intermediate, (1R, 5S, 6S) -C (6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl)] since the discovery of useful therapeutic indications for the thio-6 - [(R) -1-hydroxyethyl] -1-methylcarba-5 penem-3-carboxylate compound. Despite research efforts to find a way to (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl)] thio- To prepare 6 - [(R) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate 10 without passing through a short-term intermediate, attempts have so far failed. An obvious method is (1R, 5S, 6S) -2- (67-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl)] thio-6 - [(R) 1-Hydroxyethyl] -1-methylcarbapenem-3-carboxylate is disclosed in Merck EP Patent Application 168,707 (1986), which is described in detail in Kumagai et al., Patent 4,866,171, Column 1, pages 22. -49. The EP application discloses a drawing of a 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium halo-20 genide compound, which is a key compound for synthesis in the European application. The European application does not mention the synthesis of 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium halide and does not mention the method to be used by experts. To synthesize 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] - [1,2,4] triazolium halide.

DE-0S 2,526,358 and 3,017,875 and U.S. Patent 4,249,934 describe urazole derivatives which are structurally different from the pyrazolo [1,2-a] triazole derivatives of the present invention.

The intermediate of the present invention provides a new and improved simple process for (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] tri -azolium-6-yl)] thio-6 [(R) -1-hydroxyethyl] -1-methylcarban-35-bapenem-3-carboxylate using a key mercapto, 6,7-dihydro-6-mercapto-5H-pyrazolo - * · [1,2-a] [1,2,4] triazolium derivative.

5,9962 In this method, (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl)] thio-6 is prepared. -1 (R) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate having the formula 5

Hy H H yHs CH> 5 l \ a) 10 ° C or a pharmacologically acceptable salt thereof, by reacting a compound of formula (II) HO CHa

»M

ce> Ttpf ς / N: oor * wherein R1 is a carboxyl protecting group and R1 is an acyl group, to react 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2 , 4] with a triazolium derivative X © ”25 N (III) 0 wherein X is an anionic charge, and thus a compound of the following formula 30

Jl! Λ> - <ΤΛ "* CH3 = -Γ il Φ u C00R1

«I

35 6 93962

Q

wherein Rx and X: 11a have the same meanings as above, and then the resulting compound of formula (IV) is subjected to carboxyl deprotection to give a carbapenem compound of formula (I).

The present invention relates to a 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivative of the formula HS "CnO X® (III) θ 0 and wherein X is anionic charge.

This compound of formula (III) is a key mercaptor reagent which serves as a starting material in the synthesis described above.

The invention also relates to a process for the preparation of a compound of formula (III) by reacting a pyrazolidin-4-yl disulphide -KE of the following formula with a formimidic acid ester to give 6,7-dihydro-5H -pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulfide (Xx) wherein the X has the same meaning as above, and then the resulting formula of formula (V) is reduced. to give a compound of formula (III).

li 7 93962

The significant properties of the carbapenem compound (I) are that the 2-position substituent of the carbapenem backbone is a 6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-ylthio group and that the compound has a better antibacterial -5 n effect and resistance to DHP.

The carbapenem compound of formula (I) can be prepared according to the method described below.

The carbapenem of formula (I) can be prepared by reacting pyrazolidin-4-yl disulfide 15 - (s-CS) 2 of the following formula with a formimidic acid ester to form a 6,7-dihydro -5H-pyrazolo [1,2-a] [1,2,4] triatsolium-6-yylidisulfidi

- ( "- CDO

wherein X is an anionic charge, (b) reducing the resulting compound of formula (V) to form 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1 , 2,4] triazolium derivative HS-C 2 F> 1 X © (111) Θ 35 8 93962

Q

wherein X: 11a has the same meaning as above, (c) reacting the resulting compound of formula (III) with a compound of the following formula 5 HO γη, T 2 B? Von »CH, ^ | - (II) K».

wherein R1 is a carboxyl protecting group and Ra is an acyl group to give a compound of the following formula HO "CH3 * H H. X © uv)

h0J-L-K

u COOR1 wherein R1 and X® have the same meanings as above, (d) the resulting compound of formula (IV) is then deprotected with a carboxyl protecting group to give a carbapenem compound of formula (I). 25 • · H2 H H γΗί

Il f / S- <I> (I) CH3 = 1-1 if H J_J, _li 0 30 0 ^ ~ N Sooe 9 93962

In the present patent application, the term "lower", which defines a group of a compound, means a group or compound having 1 to 7, preferably 1 to 4, carbon atoms in this definition.

As used herein, the term "lower alkyl" refers to a straight or branched chain hydrocarbon group preferably having 1 to 6 carbon atoms, and may include, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, isobutyl, sec- butyl, tert-butyl-10, n-pentyl, isopentyl, n-hexyl, isohexyl or the like.

As used herein, the term "carboxyl protecting group" refers to any group that can protect the carboxyl group of a given compound without adversely affecting any other substituent or subsequent reactions, and such groups may include, for example, an ester residue such as a lower alkyl ester residue, including for example, methyl ester, ethyl ester, n-propyl ester, isopropyl ester, n-, iso-, sec-20 or tert-butyl ester, n-hexyl ester or the like; an aralkyl ester residue such as benzyl ester, n-nitrobenzyl ester, o-nitrobenzyl ester, p-methoxybenzyl ester or the like; and a lower aliphatic acyloxymethyl ester residue such as acetoxymethyl ester, propionyloxymethyl ester, • n- or iso-butyryloxymethyl ester, pivaloyloxymethyl ester or the like.

As used herein, the term "acyl group" means, more narrowly, a moiety obtainable by removing a hydroxyl group from the carboxyl group of an organic carboxylic acid, as well as, more broadly, any acyl group derived from an organic sulfonic acid or an organic phosphoric (5) acid. Such acyl groups may be, for example, a lower alkanoyl group such as acetyl, propionyl, butyryl or the like, a (halogen) lower alkylsulfonyl group such as methanesulfonyl, trifluoromethanesulfonyl or the like; a substituted or unsubstituted arylsulfonyl group such as benzenesulfonyl, p-nitrobenzenesulfonyl, p-bromobenzenesulfonyl, toluenesulfonyl, 2,4,6-triisopropylbenzenesulfonyl or the like; and diphenylphosphoryl.

As used herein, the term "amino protecting group" refers to groups commonly used in peptide chemistry, for example, aromatic acyl groups such as phthaloyl, benzoyl, benzoyl substituted with halogen, nitro, or lower alkyl of 1 to 4 carbon atoms (e.g., chlorobenzoyl, p-nitrobenzoyl). , p-tert-butylbenzoyl, toluoyl), naphthoyl; phenylacetyl; phenoxyacetyl; benzenesulfonyl; benzenesulfonyl substituted with lower alkyl of 1 to 4 to 20 carbon atoms (e.g., p-tert-butylbenzenesulfonyl, toluenesulfonyl); acyl derived from an aliphatic or halogenated aliphatic carboxylic acid such as acetyl, valeryl, capryl, n-decanoyl, acyl-yl, pivaloyl, haloacetyl (e.g. monochloroacetyl, monobromoacetyl, dichloroacetyl), trichloroacetyl; kamforsulfonyyli; methanesulfonyl; esterified carboxyl groups such as ethoxycarbonyl, tert-butyloxycarbonyl, isobornyloxycarbonyl, phenyloxycarbonyl, trichloroethoxycarbonyl, benzyloxycarbonyl, etc .; carbamoyl groups such as methylcarbamoyl, phenylcarbamoyl, naphthylcarbamoyl, etc. and the like thiocarbamoyl groups.

As used herein, the term "anion charge" refers to a counter anion charge for a quaternary ammonium cation charge, and such may include, for example, 11,93962 hydroxy anion; alkoxyanion such as methoxyanion, ethoxyanion; halogen anion such as chloride anion, bromide anion, iodide anion, fluoride anion; and the "acid anion residue" described below.

The term "acid anion residue" as used herein may include an acidic anionic moiety obtainable by removing a hydrogen atom from an organic acid, for example a lower fatty acid such as acetic acid, propionic acid, butyric acid or trifluoroacetic acid; a substituted or unsubstituted aryl acid such as benzoic acid, p-nitrobenzoic acid or the like; a (halo) lower alkylsulfonic acid such as methanesulfonic acid Posta or trifluoromethanesulfonic acid; a substituted or unsubstituted arylsulfonic acid such as Bent-15 sulfonic acid, p-nitrobenzenesulfonic acid, p-bromobenzenesulfonic acid, toluenesulfonic acid, 2,4,6-triisopropylbenzenesulfonic acid or the like; a substituted or unsubstituted arylphosphoric (5) acid such as diphenylphosphoric (5) acid; And inorganic acids such as nitric acid, nitric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, perchloric acid, fluorobromic acid or the like.

Mercapto reagent of formula (III), 6,7-dihydro-25-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triacat solium derivative used as a starting material in a process for preparing a carbapenem compound of formula (I) can be prepared according to the following reaction scheme A.

12 93962

Reaction Scheme A

/ ^ N-R2 s HS_V - n-r3 \ <a) (x> “fs ^ OS) 2" AN-R2 (XII) VN-r3 10 (XI) / NR ^ 0CH = NH ^ 15 -fs- C'O (xiii)> -fs-τηΛ 2xe

v ^ nh y2 (d) ^ A

W (v) 20 ;; —-> hs - / ^ TA x © (e) N-H * 5® / 25 ©

Cm) wherein R 2 and R 3 are each independently a hydrogen atom or an amino protecting group (R 2 and R 3 are not both hydrogen); R * is a lower alkyl group; and Y is an acid anion residue.

Step (a) comprises an oxidation reaction of a 4-mercapto-pyrazolidine derivative of formula (X) to give a pyrazolidin-4-yl disulfide derivative of formula (XII).

Il 13 93962

The oxidation reaction of the 4-mercaptopyrazolidine derivative of the formula (X) can be carried out in a manner known in organic chemistry as the oxidation reaction of thiol compounds. For example, the reaction may be carried out in a reaction-inert solvent such as chloroform, dichloromethane or the like in the presence of an oxidizing agent such as hydrogen peroxide, peracid, copper (II) chloride, bromine, iodine or halosuccinimide. The air oxidation reaction (in the presence of a base) can also be used for this purpose. Preferred reaction conditions are obtained by using iodine or air oxidation as an oxidant in the presence of iron powder.

Step (b) comprises a reaction in which a 4-substituted pyrazolidine derivative of formula (XI) is reacted with a sulfur compound to give a pyrazolidin-4-yl disulfide derivative of formula (XII).

The acidic anion residue represented by the symbol "Y" in the compound of formula (XI) is an acidic moiety obtainable by removing a hydrogen atom from the above acid, and preferably Y may be a halogen atom such as chlorine or bromine; or a sulfonic acid residue such as a methanesulfonyloxy residue, a toluenesulfonyloxy residue or a trifluoromethanesulfonyloxy residue.

The sulfur compound used in this reaction may be sulfur powder, hydrogen sulfide, sodium sulfide or the like.

Step (c) is a step in which the amino protecting groups, R 2 and R 3 are removed from a compound of formula (XII) obtained from step (a) or (b) above to give a pyrazolidine-4-formula of formula (VI). yylidisulfidia.

Deprotection of R2 and R3 can be carried out by a deprotection reaction known per se, such as solvolysis or hydrogenation. In a typical reaction, a compound of formula (XII) may be treated, for example, in a solvent mixture such as tetrahydrofuran-water, tetrahydrofuran-ethanol-water, dioxane-water, dioxane-ethanol-water, n-butanol-water or the like, containing morpho-Lino-propanesulfonic acid hydroxy buffer solution (pH 7), phosphate buffer solution (pH 7), dipotassium phosphate, sodium bicarbonate or the like, using hydrogen at pressures of 1 to 4 atmospheres, a hydrogenation catalyst such as palladium on carbon, or platinum oxide, in the presence of palladium hydroxide activated carbon, at a temperature range of about 0 ° C to about 50 ° C, with a reaction time of about 0.25-4 hours.

As a result, the compound of formula (VI) can be obtained as a di- or tetra-acid addition salt by treating the compound with an acid, and possible acid addition salts include: pyrazolidin-4-yl disulfide di- or -tetrahydrochloride salt, di- or tetrahydrobromide salt, di- or a tetrahydroiodide salt, a di- or tetra-trifluoroacetic acid salt, a di- or tetra-trifluoromethanesulfonic acid salt and a di- or tetra-toluenesulfonic acid salt.

Step (d) comprises the reaction between a pyrazolidin-4-yl disulfide of formula (VI) and a formimidic acid ester derivative of formula (XIII) to give a 6,7-dihydro [1,2-a] [ 1,2,4] triatsolium-6-yylidisulfidia.

The reaction may be carried out by reacting a compound of formula (VI) with a formimidic acid ester derivative such as ethyl formimidate hydrochloride, methylformimidate hydrochloride or benzylformimidate hydrochloride in an inert solvent such as water, alcohol, tetrahydrofuran or acetone. The pH of the reaction medium can be adjusted to about 6-8 by adding a base such as sodium hydrogencarbonate, potassium hydrogencarbonate, sodium carbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, sodium acetate, potassium acetate or the like base.

15 93962

The amount of the formimide acid ester derivative of the formula (XIII) is not critically accurate, and may suitably vary within a range usually of about 1 to 20 moles, preferably 4 to 12 moles, per one mole of the compound (V). The reaction temperature is not limited to certain limits, and may range from about -78 ° C to about room temperature, preferably from about -20 ° C to about 10 ° C. The reaction can be terminated under such conditions as described above, usually from about 5 minutes to about one hour.

6,7-Dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulfide of formula (V) can be obtained from the reaction mixture in crystalline form in this reaction.

Step (e) is a step for preparing a 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivative of formula 15 (III) by reduction of formula (V) compound obtained in the previous step (d).

The reduction reaction can be carried out under known reaction conditions used to cleave the sulfur-sulfur bond of a disulfide compound, for example, by reduction with a trialkylphosphine such as trimethylphosphine, triethylphosphine, tributylphosphine; triarylphosphine such as triphenylphosphine; metal reduction; or by reduction with sodium borohydride, lithium aluminum hydride or lithium triethylborohydride. In particular, the reduction with tributylphosphine can be advantageously used. The reaction is preferably carried out in an inert solvent such as water; in an alcohol such as methanol, ethanol or isopropanol; In an ether such as ethyl ether, tetrahydrofuran or dioxane; or in a mixture of these solvents in the presence of a reducing agent.

The reaction time and temperature are not limited to certain limits and may vary depending on the reducing agent used. Typically, the reaction temperature is in the range of about -20 ° C to about 50 ° C, and the reaction time may be from about 10 minutes to about 10 hours. After the reduction reaction, the 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivative of the formula (III) can be obtained as a quaternary ammonium salt of the formula (III) using known methods such as distillation, extraction, lyophilization, etc.

6,7-Dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium chloride, bromide, iodide, trifluoroacetate-10'-methanesulfonate or p-toluenesulfonate can be obtained by the above reaction.

(1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl)] thio-6 of formula (I) - [(R) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate can be prepared according to the following Reaction Scheme B.

II

17 93962

Reaction Scheme B

CH HS ^^ I ^ NX ° 5 ϊ γΗ = ^ 0 ° Ra · "cm) οη, ^ ΤΤ V -> nj — b — if) 0 XOOR1 (Π) 10 Η» HH £ Hä / ~ s ^ u X ® »WV —- * ν ^ ™, * (IV) H2 HH» Hj / "N" \ 20 ^ Αΐτγ5 <^ nrr — N \ Θ

V COO

wherein R1, R * and X® have the same meanings as above.

Step (f) comprises a reaction in which a compound of formula (II) is reacted with a mercapto reagent, a 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] - [1,2,4] triazolium derivative , of formula (III), to give a compound of formula (IV).

The reaction of a compound of formula (II) with a mercaptor reagent of formula (III) may be carried out, for example, by reacting a compound of formula (II) with a mercaptor reagent of formula (III) in excess of about an equimolar amount of 93962 18 to about 1.5- in a suitable molar amount, in a suitable solvent such as tetrahydrofuran, dichloromethane, dioxane, dimethylformamide, dimethyl sulfoxide; acetone in triil, hexamethylene phosphoramide or the like, preferably in the presence of a base such as sodium hydrogen carbonate, potassium carbonate, triethylamine, diisopropylethylamine or the like at a temperature of about -40 ° C to about 25 ° C. with a time of about 30 minutes to about 24 hours.

The reaction described above gives a carbapenem compound of formula (IV) in which the carboxyl group in its 3-position is protected by a carboxyl protecting group R1. The deprotection, R1, can be carried out by a deprotection reaction known per se, such as solvolysis or hydrogenation. In a typical reaction, the compound of formula (IV) may be treated, for example, in a solvent mixture such as tetrahydrofuran-water, tetrahydrofuran-ethanol-water, dioxane-water, dioxane-ethanol-water, n-butanol-water or the like in a solvent mixture containing morpholinopropanesulfonic acid sodium hydroxide buffer solution (pH 7), phosphate buffer solution (pH 7), dipotassium phosphate, sodium bicarbonate or the like, using hydrogen at pressures of 1 to 4 atmospheres, a hydrogenation catalyst such as palladium on carbon, platinum oxide, or platinum oxide. in the presence of carbon-activated carbon, at temperatures of about 0 ° C to about 50 ° C, with a reaction time of about 0.25 hours to 4 hours.

The result is the preparation of 30 (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl of formula (I) )] thio-6 - [(R) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate.

The carbapenem compound of formula (I) is highly stable against dehydropeptidase (DHP) and has a better antibacterial effect.

»» Li 19 93962

The preparation of the intermediates (III) of the present invention and their use in the preparation of the carbapenem compounds (I) are described in more detail by the following examples.

5 The symbols used in the following description have special meanings.

Ac: acetyl group Z: benzyloxycarbonyl group PNB: p-nitrobenzyl group 10 Example 1 nh2nh2-h2o-> NH2NHCHO

Cl) (2) 15 / CH3 -> OHCNHN = \ C3) XcH

To a solution of 377 g of hydrazine monohydrate in 760 ml of ethanol was added dropwise 726 ml of ethyl formate at 0 ° C over 1 hour, and the reaction mixture was stirred for 30 minutes at the same temperature and 14 hours at room temperature. The reaction mixture was then added over 30 minutes to 1011 ml of acetone, and the resulting mixture ·; The mixture was stirred for 30 minutes at room temperature. After the reaction, the solvent was removed under reduced pressure to give 721 g (96%) of compound (3) as a white solid.

NMR (CDCl 3) δ: 1.92 (s, 3H), 2.00 (s, 3H), 8.65 (d, 1H, J = 9.9 Hz) Example 2 / CH 3 OHCNHN = (-> OHCNN = (XCH3 xch3 35 C3) (4) «20 93962

To a solution of 1 g of compound (3) in 10 ml of methanol was added 2.12 g of a 28% sodium methoxide-methanol solution at room temperature, and the mixture was stirred for one hour and then heated to reflux for 30 minutes. Then, 1 ml of allyl bromide was added to the reaction mixture at room temperature, and the mixture was stirred for one hour at the same temperature, then heated to reflux for 30 minutes. After the reaction, the mixture was cooled to room temperature and neutralized by adding formic acid. After the reaction solvent was removed under reduced pressure, the resulting residue was purified by column chromatography to give 933 mg (67%) of compound (4) as a colorless oil.

NMR (CDCl 3) δ: 1.86 (s, 3H), 2.13 (s, 3H), 4.08-4.24 (m, 2H), 5.17-5.37 (m, 2H) Δ 5.66-5.91 (m, 1 H), 7.93-8.07 (m, 1 H)

Example 3

OHCNN - χ -3 »0HCNNH 2 · HC0 2 H

XCH '(4) (5) 3.7 g of compound (4) was dissolved in 37 ml of formic acid, and the solution was stirred for six hours at room temperature. After the reaction, the solvent was removed under reduced pressure, and the residue was purified by column chromatography to give 3.1 g (80%) of compound (5) as a pale yellow oil.

NMR (CDCl 3) δ: 1.79 (brs, 1H), 4.1-4.3 (m, 2H), 5.2-5.5 (m, 2H), δ 5.6-6.0 ( m, 1 H), 7.9-8.4 (m, 3 H)

Example 4

Br 35 ohcnnh2-hco2h -> ohcnnh2 (5). (6) k <\

II

21 93962

To a solution of 3.1 g of compound (5) in 15 ml of dichloromethane was added a solution of 2.2 g of sodium bromide monohydrate in 6 ml of methanol. To this mixture was added dropwise over 30 minutes under ice-cooling 2.16 ml of bromine in 5 ml of dichloromethane, and the resulting mixture was stirred for 30 minutes at room temperature. After the reaction, 6 ml of an aqueous solution of 7.1 g of NaHCO 3 and 5 ml of saturated aqueous sodium sulfite solution were added to the reaction mixture, and it was stirred for 10 ml to 10 minutes under ice-cooling. The mixture was extracted with ethyl acetate and the organic layer was dried over Na 2 SO 4. After the solvent was removed under reduced pressure, the resulting residue was purified by column chromatography to give 5.24 g (95%) of compound (6) as a colorless oil. NMR (CDCl 3) δ: 1.96 (brs, 1H), 3.4-4.5 (m, 5H), 8.0-8.4 (m, 2H) Example 5 Br

'v n / ^ NH

OHCNNH, -> Br \ N-CHO (6) (7) 5.5 g of K2CO3 were added to an ice-cooled solution of 5.2 g of compound (6) in 26 ml of acetone-25 rill, and the mixture was stirred for one hour. at room temperature. After the precipitate was removed, the solvent was removed under reduced pressure. The residue was purified by column chromatography to give 3.2 g (90%) of compound (7) as a pale yellow oil.

NMR (CDCl 3) δ: 1.68 (brs, 1H), 3.8-4.6 (m, 5H), 8.50 (s, 1H) Example 6

Br "O-cHO CH3cos ~ OiHCHo 35".

™ (8) 22 93962

To a solution of 3.0 g of compound (7) in 15 ml of acetonitrile was added 2.4 g of potassium thioacetate, and the mixture was stirred for 5 hours at room temperature. After the precipitate was removed by filtration, the solvent was removed under reduced pressure. The resulting residue was purified by column chromatography to give 2.65 g (91%) of compound (8) as a pale yellow oil.

NMR (CDCl 3) δ: 1.61 (brs, 1H), 2.37 (m, 3H), 3.4-4.2 (m, 5H), 8.24 (s, 1H). »Scr® 15 (8) (9)

To a solution of 2.5 g of compound (8) in 10 ml of dichloromethane: methanol (4: 1). 2.9 g of a 28% sodium methoxide-methanol solution were added under ice-cooling, and the mixture was stirred for 5 hours at the same temperature. Formic acid was then added to the reaction mixture for neutralization, and the precipitate was removed by filtration. The solvent was removed and the resulting residue was purified by column chromatography to give 1.7: 25 g (90%) of Compound (8) as a pale yellow oil.

NMR (CDCl 3) δ: 1.83 (d, 1H, J = 5.3 Hz), 2.8-3.0 (m, 2H), 3.2-3.4- (m, 2H), δ .9-4.1 (m, 1H); 8.45 (s, 1H)

Example 8: 30 hs-OTcho) \ V-N-CHO / 2 C9) (10) 35

II

23 93962 1.7 g of compound (9) was dissolved in a mixed solution of 5.1 ml of methanol: water (2: 1). To this solution were added 2.6 g of KHCO 3 and 340 mg of Fe (II) C 12.6H 2 O, and the mixture was stirred overnight at room temperature. After the precipitate 5 was removed, the solvent was removed under reduced pressure, and the residue was purified by column chromatography to give 1.5 g (90%) of compound (10) as a colorless oil.

NMR (CDCl 3) δ: 4.0-4.3 (m, 2H), 5.2-5.5 (m, 2H), 5.6-5.9-10 (m, 1H), 8.0 -8.3 (m, 2H)

Example 9 / N-CHO. // "N-CH0 \ \ __ N-CH0 \ \ __ N-CH0 / 2 15 CIO (12)

To a solution of 8.12 g of compound (11) in 75 mL of methanol: water (2: 1) was added 10.2 g of KHCO 3 20 and 1.37 g of Fe (II) C 12 · 6h 2 O, and the mixture was stirred for 8 hours at room temperature. After the reaction, the precipitate was removed by filtration, and the solvent was concentrated under reduced pressure. The residue was dissolved in chloroform and dried: sodium sulfate. The solvent was removed under reduced pressure to give 7.25 g (90%) of compound (12) as a colorless oil.

NMR (CDCl 3) δ: 3.10-4.00 (m, 8H), 4.55-4.75 (m, 2H), 8.42 (s, 4H) Example 10 i3 ° 4sY "TCHO) -> ) -2HCl V VN-CHO / 2 \ v-NH / 2 (12) 03) 24 93962 7.25 g of compound (12) was dissolved in a mixed solution of 11.4 ml of concentrated HCl and 103 ml of methanol, and the mixture After stirring for 6 hours at room temperature, the precipitate was collected by filtration and dried in vacuo to give 5.08 g (85%) of compound (13) as a white solid.

NMR (CDCl 3) δ: 3.39 (dd, 4H, J = 3.8 Hz, 13.0 Hz), 3.59 (dd, 4H, J = 6.76 Hz, 13.0 Hz), 3, 94 to 4.02 (m, 2H)

Example 11 10 HS- (TZ -> -fS ") V - N-Z \ V

To a solution of 11.4 g of compound (14) in 20 ml of dichloromethane were added 0.56 ml of triethylamine and 508 mg of iodine, and the reaction mixture was stirred for 10 minutes at room temperature. After the reaction, the mixture was washed with sodium thiosulfite solution and saturated sodium chloride solution, and then dried over sodium sulfate. After the solvent was removed under reduced pressure, the resulting residue was purified by silica gel column chromatography to give 939 mg (85.2%) of compound (15) as a pale yellow solid.

NMR (CDCl 3) δ: 3.25 (m, 2H), 3.40 (m, 2H), 3.70 (m, 2H), 4.10- (m, 4H), 5.16 (s, 8H ), 7.30 (2.20H)

Example 12 -f), - -K2 i »(15) (16) 35 • · 25 93962

A mixture of 742 mg of compound (15) in 3.1 g of a hydrobromic acid-acetic acid solution (25%) was stirred for 4 hours at room temperature. The precipitate formed after the reaction was collected by filtration and washed with 10 ml of ethyl acetate. This precipitate was treated with methanol to give 420 mg (79%) of compound (16) as a pale brownish solid.

NMR (D 2 O) δ: 3.50 (4H, m), 3.53-3.85 (6H, m)

Example 13 10 {) 2 -4HBr i s-CD X · 2cie l © 06) 07) 15 - * {s “CCn) 2 · 2οΡϊεοοθ Θ • * 0 (18) 852.2 mg of KHCO 3 was added to an ice-cooled solution. , containing 568 mg of compound (16) in 30 ml of water, to adjust the pH of the solution to 7.10. To this solution was then added 2.112 g of formimidate hydrochloride, and the reaction mixture was stirred for 10 minutes under the same conditions. After adjusting the pH of the reaction mixture to 5.5 with 1 N HCl, the mixture was washed with 50 mL of ethyl acetate. The aqueous solution was concentrated to dryness under reduced pressure, and 30 ml of methanol was added to the residue. After the precipitate was removed by filtration, the solvent was removed, and the resulting residue was purified by SP-207 column chromatography, and the eluent was lyophilized to give compound (17).

• · 26 93962

The compound (17) obtained in the above lie was dissolved in 10 ml of methanol and treated with 1 ml of trifluoroacetic acid. The solvent was removed under reduced pressure to give 317.2 mg (58.3%) of compound 5 (18) as white crystals.

NMR (D 2 O) δ: 4.80-5.00 (4H, m), 5.00-5.16 (6H, m), 9.05 (4H, s)

Example 14 10 (s-CD) * 2cf3co ° 0 - ^ hs ^ j! 3n (18) CF3C00 © 15 (19)

To a solution of compound (18) in 6 ml of water-tetrahydrofuran (1: 1) was added 0.082 ml of tri-n-butylphosphine under ice-cooling, and the mixture was stirred for one hour at the same temperature. After the reaction, 10 ml of water was added to the reaction mixture, and the mixture was washed with dichloromethane and ethyl acetate. The aqueous layer was lyophilized to give 102.2 g (90.9%) of compound (19) as a white solid.

NMR (D 2 O) δ: 4.50-4.70 (2H, m), 5.00-5.20 (3H, m), 9.00 (2H, s) 27 93962

Example 15 5 HO H H CH 2 ° C [μl] 0P (0Ph) 2 HS— 'CF = C0 ° θ cr n ^ COOPNB φ (20) (19) 10 HO It u CH3 J — N- ^ φ

O ^ COOPNB

(21) To an ice-cooled solution of 85.5 mg of compound (19) and 199.6 mg of compound (20) in 3 ml of anhydrous acetonitrile was added dropwise 0.06 ml of diisopropylethylamine, and the mixture was stirred for one hour at the same temperature. After the reaction, the solvent was removed under reduced pressure, and the resulting residue was dissolved in 20 ml of ethyl acetate, and then the solution was centrifuged at 3000 rpm for 25 minutes. After centrifugation, the supernatant was decanted off. Centrifugation and decantation were repeated three times, and each time the liquid above the precipitate was collected and concentrated under reduced pressure. The resulting residue was dissolved in 20 ml of chloroform, and the mixture was centrifuged, and the supernatant was decanted off. Each residue obtained in the above centrifugation was collected and then dissolved in methanol. Removal of methanol under reduced pressure gave • 28 93962 134.8 mg (67%) of compound (21) as a pale yellow oil.

NMR (CD 3 OD) δ: 1.32 (d, 6H, J = 6.0 Hz), 3.35 (m, 1H), 3.65 (m, 1H), 4.20 (m, 1H), δ .42 (m, 1H), 4.60-4.90 (m, 2H), 5.1-5.3 (m, 3H), δ 5.36 (Abq, 2H, J = 13.7 Hz) , 7.67 (d, 2H, J = 8.5 Hz), 8.21 (d, 2H, J = 8.5 Hz), 9.07 (s, 1H), 9.08 (s, 1H)

Example 16 jjjx ^ s-cry · 10 cH = yxx ® 0 "N '" COOPNB (21) 15 "rhh tHi M * (22) 20 To a solution of 84.9 mg of compound (21) in 2.5 ml of 0, In 1 M acetate buffer and 2.5 ml of n-butanol, 25 mg of 10% palladium-on-carbon was added, and the catalytic hydrogenation was performed at room temperature for one hour at a pressure of 4.0 atm. removed, washed with 20 mL of water, the solvent was combined, and the pH of the solution was adjusted to 4.8-5.6 by the addition of 1 N NaOH, and washed with 20 mL of n-butanol, and the solvent was concentrated to 5 mL under reduced pressure. , and the resulting residue was purified by SP-207 column chromatography and recrystallized from ethanol to give 30 mg (60%) of compound (21) as white crystals.

NMR (D 2 O) δ: 1.29 (d, 3H, J = 7.3 Hz), 1.33 (d, 3H, J = 6.3 Hz), 3.44 (dq, 1H, J = 7, 3, 9.5 Hz), 3.56 (dd, 1H, J = 2.9, 6.2 Hz), 35 4.30 (quintet, 1H, J = 6.2 Hz), 4.34 (dd , 1H, J = 2.9, 9.5 Hz), 4.75-4.84 (m, 2H), 5.08-5.17 (m, 2H), 4.98-5.04 (m , 1H), 9.06 (s, 1H), 9.07 (s, 1H)

Claims (3)

29 93962 1. 6,7-Dihydro-6-mercapto-5H-pyrazolo [1,2-a] - [1,2,4] triazolium derivative of the formula (III): 5 HS-^ O ^ (III> Φ 10 where X- is the anlon charge. Process for the preparation of 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivative (III) according to Claim 1, characterized in that 6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulfide of formula (V)? 2X Θ V ν-'Ν-Ξ- / J (V) 20. y2 wherein X "is an anlonic charge to give a compound of formula (III). The method according to claim 1, 6,7-! For the preparation of dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivative (III), characterized in that a pyrazolidin-4-yl disulfide of formula (VI) is obtained. 93962 or a salt thereof to react with a formimidic acid ester to give 6,7-dihydro-5H-pyrazo-Solo [1,2-a] [1,2,4] triazolium-6-yl disulfide of formula (V) x 0 wherein X1 is an anionic charge, and then the resulting compound of formula (V) is reduced to form a compound of formula (lii). % »II · 31 93962 1. 6,7-Dihydro-6-mercapto-5H-pyrazolo [1,2-a] - [1.2.4] triazolium derivatives of formula (III): 5 hs-ΟΓΛ X® (1II> ® 10 for X 'is enionated. A process for the preparation of 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivatives (III) according to claim 1, which is incorporated herein by reference. reducing agents of 6,7-dihydro-5H-pyrazolo [1,2-a] - [1.2.4] triazolium-6-yl disulphide with the formula (V) 20 - (S “CpN) 2 2X @ by X” , varvid erhälls en förening med formeln (III). 25 3. A process for the preparation of 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivatives (III) according to claim 1, which is described in the invention. pyrazolidin-4-yl disulfide with formyl (VI) 30 '35 32 93962 or with salt of formimide syrup, 6,7-dihydro-5H-pyrazolo [1,2-a] [1,2 , 4] triazolium-6-yl disulfide with formula (V) ^ -03 ") 2, and 10 with X 'is an anionized, and then reduced with the compounds of formula (V), colors with compounds of formula (III) .li