FI95382C - 6,7-dihydro-5H-pyrazolo / 1,2-a // 1,2,4 / triazolium-6-yl disulfide, process for preparing it using pyrazolidin-4-yld disulfide and pyrazolidin-4-yld disulfide - Google Patents

Description

95382 6.7-Dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulfide, process for its preparation using pyrazolidin-4-yl disulfide and pyrazolin-4-yl disulfide

Divided by application 933,245 (Patent 5,93962)

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] triazolium of formula (I) -6-10 yl)] thio-6 - [(R) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate

HTH

CHa ^ i 1-1 II

is HJ -— N \ Θ ® (I) 0 C00 or a pharmacologically acceptable salt thereof. The invention also relates to the preparation of intermediates.

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-yl] -1-methylcarbapenem-3-carboxylate as a carbapenem antibiotic, see U.S. Patent 4,866,171, 4,925,836 and 4,925,935, this compound being carba-2-penem-3- a carboxylic acid compound in which the β-methyl group is attached at the 1-position and a 6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-ylthio group represented by the following formula 2 95382 -s- <T> Φ 5 is attached to the 2-position in the carbapenem backbone Due to these certain substitutions, the compound of formula (I) has a better antibacterial activity and is chemically and physicochemically very stable in the body and stable against dehydropept-10 dase (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] -1-methylcarbapenem-3-carboxylate 15 Kumagain et al. in the publication, occurs from the imino intermediate. According to Eli Kumagai et al., A compound of formula (I) can be prepared by reacting a compound of formula (II): CH 2 OH (S)

CHa ΰ I I

ς / N XttOR1 25 wherein R1 is a carboxyl protecting group and R * is an acyl group, to react with a mercaptor reagent of formula (VII): / —N-Rb HS— (| (VII) λ-N-Rb wherein Rb is an amino protecting group, and thus a compound of formula (VIII): 35 3 95382 H? H - f ”“ N-Rb ^ COOR 'wherein R1: IIS and Rb have the same meanings as above, and by carrying out protecting groups for a compound of formula (VIII), Removal of R1 and Rb gives a carbapenem compound of formula (IX)

Hw H H SH3 / —NH

U C00H

and then, by reacting the resulting compound of formula (IX) with a formimidic 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: 30

H ° H H 2 H 3 N-CH = NH

CH ^ *

H I N-CH = NH

J-N: \

35 0 C00H

4,9382 This imino intermediate is a non-stable compound and undergoes a cyclization reaction to form a compound of formula (I).

One method of (1R, 5S, 6S) -2 - [(6,7-dihydro-5H-pyraz-5-solo [1,2-a] [1,2,4] triazolium-6-yl)] thio-6 - for the preparation of [(R) -1-hydroxyethyl] -1-methylcarbapenem-3-carboxylate is described in Merck EP Patent Application 168,707 (1986), which is described in detail in Kumagain et al., Patent 4,866,171, column 1, pages 22- 49. EP-10 discloses a drawing of 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium halide, 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-15 halide and does not mention the method for the use of 6,7-dihydro-6 -merkap-to-5H-pyrazolo [1,2-a] [1,2,4] triatsoliumhalogenidinsynte-tisoimiseksi.

The invention relates to a 6,7-dihydro-5H-pyrazolo [1,2-a] -20 [1,2,4] triazolium-6-yl disulphide of formula (V): - (s-CpN) 2 2X ° < v) 25 wherein X 'is the anionic charge.

The invention also relates to a process for the preparation of said 6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulphide (V) by carrying out the pyrazolidine of formula (VI). 4-yl disulfide - (- a), <1 1H «» * · II * **> 5 95382 or a salt thereof to react with a formimidic acid ester to give a compound of formula (V).

The invention further relates to said pyrazolidin-4-yl disulfide (VI).

Reduction of said compound of formula (V) gives a 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] - [1,2,4] triazolium derivative having the formula 10 ΧΘ (III) 0

Q

where X is the anionic charge.

This compound of formula (III) is a key mercaptor reagent which serves as a starting material for the preparation of therapeutically valuable end products of formula (I).

(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-hydroxy-ethyl] -1-methylcarbapenem-3-carboxylate 20

Hw H H -Ha I N (i)

CH / n-f | T

h ji-y ® 0 xoo ° 25 or a pharmacologically acceptable salt thereof is prepared in such a way that the compound of formula (II) Π0 CH3 f H H w „

30 A ^ 1JJK / 0R

ch3 - I if (II): ς / N xoor1 35 wherein R1 is a carboxyl protecting group and R1 is an acyl group.

6 95382 is reacted with a 6,7-dihydro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] triazolium derivative of the above formula (III) 5 ^ N ^ K1 v © HS IN 1 x (m) 0 0 wherein X is an anionic charge to give a compound of formula (IV) H »HH SHs /" N ^ X® A <1 isK ^ s- <i> (IV) CH j ^ =] —F | f VN ^ 15 "J — N— ^ ® u C00R1 wherein R1 and X6 have the same meanings as above, and then the resulting compound of formula (IV) is subjected to deprotection of the carboxyl protecting group to give the formula (IV). I) carbapenem compound.

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-30 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 similar groups; and a lower aliphatic acyloxymethyl ester residue such as Example 5, xixacetoxymethyl ester, propionyloxymethyl ester, 11 μl II »:» m tl! M I fr »1> t 7 95382 n- or iso-butyryloxymethyl ester, pivaloyloxymethyl ester or similar groups.

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 (halo) 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 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, acyloyl pivaloyl, haloacetyl (e.g. monochloroacetyl, monobromoacetyl, dichloroacetyl), trichloro; kamforsulfonyyli; methanesulfonyl; esterified carboxyl groups such as ethoxycarbonyl, tert-butyloxycycarbonyl, 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" means a counter anion charge for a quaternary ammonium cation charge, and such may include, for example, a 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 comprise an acidic anionic moiety obtainable by removing a hydrogen atom from an organic acid, for example a lower ras-waxic 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; (halogen) lower alkylsulfonic acid such as methanesulfonic acid-20 Posta or trifluoromethanesulfonic acid; a substituted or unsubstituted arylsulfonic acid such as benzenesulfonic 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.

: Mercaptor reagent of formula (III), 6,7-dihydro-

The ro-6-mercapto-5H-pyrazolo [1,2-a] [1,2,4] tritrate solium derivative used as a starting compound in the process for preparing the carbapenem compound of formula (I) can be prepared according to the following reaction scheme A

in accordance with. Compounds of formulas (V) and (VI). use as intermediates is shown in this reaction scheme.

9 95382

Reaction Scheme A

/ ^ N-R2 HS— (I o \ (a) K V-n-r3 <x). ~ fs ^ CLi3) 2 77 *, Λ * - »2. <XII) ~ V ^ N-r3 10 (XI)

/ x R ^ 0CH = NH / X

“<-C £), ^ Φ") * 2, e (VI) (V) 20 -> HS-Z ^^^ N xe (e) 25 Φ

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).

10 95382

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. 25 sulfur powder, hydrogen sulphide, sodium sulphide or. corresponding compound.

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

· The 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 tetramil nmites 11,93838 in hydrofuran-water, tetrahydrofuran-ethanol-water, dioxane-water, dioxane-ethanol-water, n-butanol in water or similar mixtures 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 1-4 atmospheres, as a hydrogenation catalyst, , in the presence of palladium-activated carbon or 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] triazolium-6-yl disulfide.

The reaction can 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 or tetrahydrofuran.

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.

95382

The amount of the formimidic 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 a compound of formula (V) the 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 the 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 triethyl borohydride. 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. Usually the reaction temperature is ra-id i- »lii m i and. . .

13 95382 at 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] triacat solium derivative of the formula (III) can be obtained as a quaternary amine of the formula (III). as the magnesium salt 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-10H, methanesulfonate or p-toluenesulfonate can be obtained by the above reaction.

In the following examples, Examples 1 to 9 and 11 illustrate the preparation of starting materials for a compound of formula (VI), Examples 10 and 12 illustrate the preparation of a compound of formula (VI) in salt form, Example 13 illustrates the preparation of compound (V) from compound (VI), Example 14 (III) for the preparation of compound (V), Example 15 for the preparation of compound (IV) for compound (III) and Example 16 for the preparation of a therapeutically valuable end product of formula (I) from compound (IV).

The symbols used in the examples have the following meanings:

Ac: acetyl group Z: benzyloxycarbonyl group 25 PNB: p-nitrobenzyl group Example 1

NH 2 NH 3 -H 2 O -> NH 2 NHCHO

. (1) (2) 30 * / CH 3 -> 0HCNHN = ((3) XCH '35 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. , and the reaction mixture was stirred for 30 minutes at the same temperature and for 14 hours at 14 ° C. At this time, the reaction mixture was added to 1011 ml of acetone over 30 minutes, and the resulting mixture was stirred for 30 minutes at room temperature. 721 g (96%) of compound (3) were obtained 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 10 CH3 0HCNHN = \ -> 0HCNN = \ XCH3 xch3 (3) C4) 15 To a solution of 1 g of compound (3) in 10 ml of methanol , 2.12 g of a 28% sodium methoxide-methanol solution was added 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 under 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), 5.66-5.91 (m, 1H), 7.93-8.07 (m, 1H) Example 3: I / CH

0HCNN = \ -> OHCNNH2 * HCO2H

xch3 35 C4) C5) 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 1 Br ohcnnh2-hco2h -> ohcnnh2 10 (5). (6)

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 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 NR / "~ nh: · OHCNNH. -> BrVJ-CH0

(6) W

5.5 g of K 2 CO 3 was added to an ice-cooled solution of 5.2 g of compound (6) in 26 ml of acetonitrile, 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 5

Br ~ O-CH0 * CH ^ C0S “OiI-CH0 <7> <»

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)

Example 7

/ 'N H v / "- NH

„Ch3cosA ^ n-cho - ^ hs-VÄ-cho 25) (8) (9)

To a solution of 2.5 g of compound (8) in 10 ml of dichloromethane: methanol (4: 1) was added 2.9 g of a 28% sodium methoxide-methanol solution on ice; under cooling, and the mixture was stirred for five 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 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), 3, 9-4.1 (m, 1H), 8.45 (s, 1H) il: UL »Iti l.iitl i 95382 17

Example 8 (HCl-fsCLol (9) OO) 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 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 (m, 1H), 8.0- 8.3 (m, 2H)

Example 9 HS-Y ^ i1 "0110 ->" f S- (? ~ CH °) 20 \ ^ N-CH0 \ V ^ N-CHO / 2 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 and 1.37 g of Fe (II) C 12 H 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 over 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 (compound of formula (VI), HCl salt) / N-CHO. 2HCl 1 S 2 VN-CHO / 2 V V-NH / 2 (12) ¢ 13) 18 95382 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 was stirred for six hours at room temperature. After the reaction, 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) V-N-Z \ V-N-Z / 2 (14) (15) 15

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 (compound of formula (vi), HBr salt)), - - (* <3 \ - (15) 06) 35 95382

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 (Compound of formula (V)) / - "NH \ _> i -2Cl ({s ^ vjJ2'4HBr ^ 06) 15 -> {s-CÖ) * Θ 20 852.2 mg KHCO 3 was added to an ice-cooled solution of 568 mg of compound (16) in 30 ml of water to adjust the pH of the solution to 7.10 To this solution was added 2.112 g of formimidate hydrochloride, and the reaction mixture was stirred for 10 minutes under the same conditions. adjusted 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 30. After precipitation was removed by filtration, the solvent was removed and the resulting residue was purified by SP-207 column chromatography. lyophilized to give compound (17).

20 9 5 3 8 2

The compound (17) obtained in the previous step was dissolved in 10 measures 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 (compound of formula (III))

io {s <X>) 2 · θοθ - * hs-CD

e φ (18) CFaCOO® (19) 15

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); »: I 95382 21

Example 15 (Compound of Formula (IV)) HO „u ch3 m _ f H H w II / ^ N ^ \ Θ

Aj = As / 0P (0Ph), HSH I, N * CF3C00 5 CH3 1 | f + (T-'N ^ COOPNB ® (20) U9) 10 Jj iX / S-TpN · CF.C00®

- * CHi ^ xxX

cr N ^ 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 five 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. After removal of methanol under reduced pressure, 95382 22 134.8 mg (67%) of compound (21) were obtained 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 (final product of formula (I))

Hw H H * Hs θ

At I> · CF3COO

10 en 3 I — T 11 J — N-®

0 ^ COOPNB

(21) 15 H2 H H 2 Hj - »« ,, ΛΰΧ.

* (22) To a solution of 84.9 mg of compound (21) in 2.5 ml of 0.1 M acetate buffer and 2.5 ml of n-butanol was added 25 mg of 10% palladium on carbon, and the catalytic hydrogenation was performed at room temperature, with a hydrogenation time of one hour, at a pressure of 4.0 atm. After removal of the catalyst, it was washed with 20 ml of water. The solvent was combined and the pH of the solution was adjusted to 4.8-5.6 by adding 1 N NaOH solution, and washed with 20 ml of n-butanol. 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; 30 mg (60%) of compound (21) were obtained 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)

23 9 5 3 8 2 1. 6,7-Dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazol-6-yl disulfide of formula (V): - {s -CPn) 2 2X ° 10 v ® where X 'is the anion charge. A process for the preparation of 6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulphide (V) according to claim 1, which is preferred for pyrazole 4-general disulfide, in the form of formula (VI), - (5_ΟΓη) 2, (vi) 20 or that the site is treated with a formimidic acid ester, the color being different from the formulation (V). Process for the preparation of 6,7-dihydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulphide (V) according to Claim 1, characterized in that ) pyrazolidin-4-yl disulfide Is -CT 2 20 V ^ · ΝΗ Λ or a salt thereof to react with a formimidic acid ester to give a compound of formula (V). A pyrazolidin-4-yl disulfide of formula (VI): -fs-cr) V \ nh y2 30 or a salt thereof. «24 9538? 1,6,7-d ± hydro-5H-pyrazolo [1,2-a] [1,2,4] triazolium-6-yl disulphide with formula (V): 5 - (φ)) θ, v> 10 for X ' is enionized. 3. Pyrazolidin-4-yl disulfide, having the formula (VI): 25 f / ^ m Λ -fS- (I <VI> y V — So y2 eller ett sait därav.