JPH05271240A - New 1-thia-4,6-diazaspiro (4,4)nonane derivative and its production - Google Patents

Abstract

PURPOSE:To obtain a new 1-thia-4,6-diazaspiro[4.4]nonane derivative useful as a preventing and treating agent for diseases caused by limited supply of the compound. CONSTITUTION:The objective compound of formula I (X is H, COOH, R<1> or COOR<1>; Y is OH, 0R<1>, A or AOR<1>; R<1> is 1-8C linear or branched hydrocarbon group; A is amino acid residue; AOR<1> is amino acid ester residue) or its pharmaceutically permissible salt, e.g. 7-carboxy-3-{[carboxymethyl)amino]carbonyl-}-1- thia-4,6-diazaspiro[4.4]nonane. This compound can be produced by subjecting a compound of formula II to III to cyclization reaction in a weakly acidic, neutral or alkaline solution and then, as necessary, converting the resultant product into its pharmaceutically permissible salt. Furthermore, the compound of formula I is useful as a preventing or treating agent for above-mentioned diseases, e.g. diabetes mellitus, arrhythmia, ulcer, angina, thrombotic diseases, carcinoma, etc.

Description

Detailed Description of the Invention

[0001]

The present invention relates to the formula [I]

[0002]

[Chemical 4]

The present invention relates to 1-thia-4,6-diazaspiro [4.4] nonane represented by: and pharmaceutically acceptable salts thereof, a process for producing these compounds and uses of these compounds.

[0004]

2. Description of the Related Art Conventionally, the formula [I '] is used.

[0005]

[Chemical 5]

The compound represented by the formula [I
I ′]

[0007]

[Chemical 6]

A compound represented by the formula [IV ']

[0009]

[Chemical 7]

It has been argued that it may exist as an intermediate during the interconversion with the compound represented by (Calvin,
M., in "Glutathione", edited by SP Colowick at a
l., Academic Press, New York, 1954, 3-3
0). Further, it has recently been argued that the radical of the compound represented by the above formula [I ′] may be generated as an intermediate when the aqueous alkaline solution of the compound represented by the above formula [IV ′] is irradiated ( Eriksen, T.
E. and Fransson, G., Radiat. Phys. Chem. 32 , 1
63-167 (1988); J. Chem. Soc. Perkin Tra.
ns. II 1117-1122 (1988)).

However, in any of these papers, no proof that the compound represented by the above formula [I '] exists certainly is given, and the compound represented by the above formula [I'] is simply identified. Nothing is said about the separation or its properties. Rather,
For the mother nucleus of the compound represented by the above formula [I '] deduced in the above Eriksen et al. Article, American Chemical Soci
ety registered with the registration number (RN. No. 1004544-83-
The addition of 9) means that the nucleus itself is new,
It shows that such a compound has not been provided yet.

In view of the problems of the prior art as described above, the present inventor has developed a novel 1-thia-4,6-diazaspiro [4.4].
The present invention has been achieved as a result of intensive research to provide a nonane derivative and a method for producing the same.

[0013]

That is, the present invention provides the formula [I]

[0014]

[Chemical 8]

[Wherein X is H, COOH, R¹, Or C
OOR¹And Y is OH, OR ¹, A, AOR¹To
Represent (where R¹Is independently C₁~ C₈Straight chain, min
Substituted by a branched or cyclic hydrocarbon group, or an aromatic group
C₁~ C_FourRepresents a linear or branched hydrocarbon group of
, A represents an amino acid residue, and AOR¹Is the amino acid
Represents a stell residue. )]
Pharmaceutically acceptable salts of
Of the formula [I] consisting of effective amounts of these compounds
Diseases caused by limited supply of compounds
Is a preventive or therapeutic agent.

It is readily understood that the compound of formula [I] has several chiral centers and therefore exists in multiple isomers. The invention extends to each of these isomers, and mixtures thereof.

In the above formula [I], X is H, COO.
H, R¹Or COOR¹(Where R ¹Is independently C₁~
C₈A linear, branched or cyclic hydrocarbon group of, or
C substituted with an aromatic group₁~ C_FourLinear or branched
Represents a hydrocarbon group). Such R¹C₁~ C
₈Examples of the linear, branched or cyclic hydrocarbon group of
For example, methyl group, ethyl group, propyl group (n-propy
Group, i-propyl), butyl group (n-butyl, i-butyl)
, S-butyl, or t-butyl), cyclohexyl
C substituted with a group or an aromatic group₁~ C_FourStraight chain
Examples of the branched hydrocarbon group include a benzyl group and a phenyl group.
Examples thereof include a nylethyl group.

Y is OH, OR ¹ , A or AOR ¹
(Wherein R ¹ independently represents a group selected from the groups defined above, A represents an amino acid residue, and AOR ¹ represents an amino acid ester residue). Examples of such OR ¹ include an alkoxy group composed of the alkyl group represented by R ¹ and an oxy group, such as a methoxy group, an ethoxy group, a propoxy group, a butoxy group, a benzyloxy group.

A is an amino acid residue such as glycine (Gly), alanine (Ala), leucine (L).
eu), threonine (Thr), aspartic acid (As
p), asparagine (Asn), methionine (Me)
t), phenylalanine (Phe), histidine (Hi
Examples thereof include those having D and L optical isomers such as s) and racemates, or β- and γ-aminocarboxylic acids. Furthermore, examples of AOR ¹ include amino acid ester residues consisting of the amino acid residue represented by A and the alkyl group represented by R ¹ .

The compound represented by the formula [I] of the present invention is
Among the above, as X, for example, COOH, C
OOR¹(Where R¹Is C₁~ C₆Straight and branched
Preferably represents a cyclic hydrocarbon group), and Y and
Then OH, OR¹(Where R¹Is C₁~ C₆Straight chain of
A branched, branched or cyclic hydrocarbon group), represented by A
A, AOR when the amino acid residue to be given is Gly
¹(Where R¹Is C₁~ C₆Linear, branched or
(Representing a cyclic hydrocarbon group) is preferable, and among them, X is
For example, COOH, COOR¹Is preferred, and Y
Is, for example, the amino acid residue represented by A is Gly
A, AOR when¹(Where R¹Is C₁~ C_Fourof
Represents an alkyl group), OH, and OR¹(Where R¹
Is C₁~ C _FourRepresents an alkyl group of
Can be listed.

The pharmaceutically acceptable salt of the compound of the present invention is, when an acidic group is produced by the X, Y substituent,
Examples thereof include addition salts with an inorganic base or an organic base, and when the compound of the formula [I] becomes basic by the X, Y substituent, addition salts with an inorganic acid or an organic acid can be mentioned.

The compound of the formula [I] of the present invention can be obtained by the following processes.

As the first manufacturing method (manufacturing method 1), the formula [II] is used.

[0024]

[Chemical 9]

[The definitions of X and Y are the same as those of the formula [I]]
The compound of formula [I] is formed by subjecting the compound of formula [I] to a ring closure reaction in a weakly acidic, neutral or alkaline solution. In some cases, the compound of formula [I] is pharmaceutically acceptable. A process for producing a compound represented by the above formula [I] or a pharmaceutically acceptable salt thereof, which comprises obtaining a salt.

As the second manufacturing method (manufacturing method 2), the formula [III] is used.

[0027]

[Chemical 10]

[The definitions of X and Y are the same as those of the formula [I]]
The compound represented by the formula (I) is subjected to a ring closure reaction in a weakly acidic, neutral or alkaline solution to form a compound represented by the above formula [I], and optionally a pharmaceutically acceptable salt of the compound of the formula [I]. A process for producing a compound represented by the above formula [I] or a pharmaceutically acceptable salt thereof.

Represented by the formulas [II] and [III],
The compound for producing the formula [I] is described in JP-A-2-121.
It can be obtained by the method described in No. 965.

In these reactions, for example, first, a strong acid salt of the compound represented by the formula [II] and the formula [III] is produced,
These compounds are then weakly acidic, neutral,
Alternatively, it is carried out by subjecting it to a ring closure reaction in an alkaline solution. As the strong acid forming the strong acid salt of the compound of the formula [II] and the formula [III], hydrochloric acid is generally used, but mineral acids such as sulfuric acid, nitric acid, perchloric acid and hydrobromic acid, benzenesulfonic acid, Organic acids such as paratoluene sulfonic acid can also be used. Further, as a base for reducing the acidity of the solution of the strong acid salt of the compound of the formula [II] and the formula [III], an inorganic base such as caustic soda, caustic potash, aqueous ammonia, an organic base such as an aliphatic amine, an aromatic amine, etc. Can be used.

The reaction conditions for such a reaction are not particularly limited, but water, a polar organic solvent or the like is used as a reaction solvent,
The reaction temperature is 0 to 100 ° C., and the reaction time is 1 minute to 24 hours.

It is also possible to prepare the compound of the formula [III] in the free form and to give the compound of the formula [I] under the above reaction conditions in a weakly acidic, neutral or alkaline solution.

Further, the formula [IV] described in JP-A-2-121965 is used.

[0034]

[Chemical 11]

[The definitions of X and Y are the same as in the case of the formula [I]]
The compound of formula [I] can be produced by isolating the compound of formula [II] or formula [III] by controlling the above reaction conditions using the compound of formula as a starting material. ..

In the compound represented by the formula [I], when the X and Y substituents give rise to acidic groups, these acidic groups can be converted into a salt form. Pharmaceutically acceptable salts include
Some are derived from inorganic bases such as sodium, potassium, calcium, magnesium, barium, lithium, ammonium and others are derived from organic bases such as diethylamine, isopropylamine, ethanolamine, choline, piperidine, arginine, lysine. .. Common salts include sodium, calcium, magnesium, ethanolamine, arginine, lysine and choline. Preferably the base addition salts are sodium and magnesium salts.

Further, the compound represented by the formula [I] has a substituted amino group having basicity. Examples of the pharmaceutically acceptable acid used when derivatizing these amino groups into salts include inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid,
Sulfuric acid and organic acids such as acetic acid, tartaric acid, maleic acid,
Citric acid, succinic acid, lactic acid, α-ketoglutaric acid, α-
Glycerophosphate and glucose-1-phosphate are included. Generally, it is an acid addition salt with an acid such as hydrochloric acid, phosphoric acid, sulfuric acid, citric acid, tartaric acid, lactic acid and acetic acid.

The compounds of formula [I] and their pharmaceutically acceptable salts are also capable of forming pharmaceutically acceptable solvates such as hydrates, which solvates also A compound of formula [I] or a salt thereof is always included when referred to herein.

The compound represented by the formula [I] of the present invention thus obtained is a compound (glutathione) wherein X is COOH and Y is HN—CH ₂ —COOH in the formula [IV] which is a natural physiologically active substance. Since it is a novel substance including a compound derived as a starting material, it is easily presumed that it exerts an important function for the living body. Therefore, as a prophylactic or therapeutic agent for various diseases caused by the fact that the supplement of the compound represented by the formula [I] is not smoothly carried out in vivo, the compound represented by the formula [I] or its pharmaceutically Acceptable salts can be used. Examples of such diseases include diabetes, arrhythmia, ulcer, angina, thrombotic disease, and cancer,
The pharmaceutical use of the compound represented by the formula [I] is not limited to these.

[0040]

EXAMPLES Next, the present invention will be specifically described with reference to examples.

[0041]

Example 1 Production of 7-carboxy-3-{[(carboxymethyl) amino] carbonyl} -1-thia-4,6-diazaspiro [4.4] nonane (Production Method 2) 9 g of glutathione in 45 ml of concentrated hydrochloric acid. After melting and heating at 100 ° C. for 3 minutes, it was immediately cooled on an ice bath. To this, 900 ml of cold ethanol was added, followed by cold ethyl acetate 4,
050 ml was added with stirring and left in the cold room for 20 minutes.
The white precipitate was collected by decanting most of the supernatant and centrifuging the rest. The collected precipitates were dried under reduced pressure over granular sodium hydroxide in a desiccator for 3 days, and further dried over powdered phosphorus pentoxide for 3 days under reduced pressure to obtain N- [2- (3-amino-3-carboxy-1). -
Propyl) -4-Δ ² -thiazolinylcarbonyl] -glycine hydrochloride (glutathione / thiazoline hydrochloride) 6.
96 g were obtained. Next, 6.41 g of glutathione / thiazoline hydrochloride obtained was cooled with 0.2 N NaOH solution 20.
The pH of the solution was quickly adjusted to 6.50 by dissolving it in 0 ml and further adding 56 ml of the same solution, and then freeze-drying was performed to obtain N- [2- (5-carboxy-2-pyrrolidinylideneamino). ) -3-Mercaptopropionyl]-
7.90 g of a crude dry product of sodium salt of glycine (glutathione / pyrrolidine) was obtained. The lyophilized glutathione-pyrrolidine was extracted with 50 ml of methanol and centrifuged to separate an insoluble fraction and a methanol solution.

The insoluble matter was extracted again with 10 ml of methanol,
After mixing with the first extract and concentrating to dryness under reduced pressure, the residue was dried under reduced pressure in a desiccator overnight to give 6.34 g.
To give a solid. Subsequently, 6.13 g of the dried solid was dissolved in 212 ml of methanol, and 141 ml of ethyl acetate was added with stirring.
Was dripped. 350 ml of ethyl acetate was similarly added to the supernatant obtained by centrifuging and removing the formed precipitate, and the formed precipitate was collected and dried over phosphorus pentoxide for 1 week under reduced pressure in a desiccator, and 3.33 g of a white solid. Got

400 mg of the glutathione / pyrrolidine Na salt solid thus obtained was dissolved in 3 ml of distilled water in a hydrolysis tube manufactured by Pierce, 7 ml of ethanol was added, and nitrogen substitution was carried out under cooling in an ice bath. 40 at 100 ° C
Heated for minutes. After the reaction, the oily substance deposited on the bottom of the tube when the container was cooled was dissolved by adding 4 ml of distilled water. This operation was repeated twice more, and a total of 1,200 mg of glutathione-pyrrolidine salt was treated to make one solution.

Next, this solution was placed in a low temperature room and QAE-Se
phadexA-25 HCO ₃ ^- type of column (2 × 32cm)
Upon elution, ammonium bicarbonate in 50% ethanol was eluted with a linear gradient of 500 ml each of 0.08M and 0.4M solutions and 16 ml fractions were collected. Each fraction was applied to a high-performance liquid chromatographic column of Inertosyl ODS-2 manufactured by GL Science Co., Ltd. and eluted with 2.0 mM potassium phosphate buffer (pH 6.6). Fractions 43 to 50 containing the desired substance were collected and concentrated to dryness under reduced pressure at 40 ° C. until a film-like substance was obtained. The residue is methanol 2
After dissolving in 0 ml, 10 ml of ethanol was added and reconcentrated.
This operation was repeated to give a white solid, which was dried under reduced pressure in a desiccator.

Of the solid thus obtained, 152 mg was dissolved in 54 ml of methanol, 300 mg of dithiothreitol was added, the mixture was replaced with nitrogen, and the mixture was allowed to stand at room temperature overnight. Was reduced. 54 ml of ethyl acetate was added thereto and the resulting precipitate was collected by centrifugation at 15,000 xg for 30 minutes. The precipitate was dissolved again in 50 ml of methanol, 74 ml of ethyl acetate was added thereto, and the precipitate was collected by centrifuging for 60 minutes and dried under reduced pressure to obtain 75 mg of 7-carboxy-3-
{[Carboxymethyl) amino] carbonyl} -1-thia-4,6-diazaspiro [4.4] nonane was obtained (S
IMS: 290 (M + H) ⁺ ).

44 mg of this purified product was dissolved in 3 ml of distilled water, neutralized with 0.1N NaOH, and freeze-dried. ¹ H-NMR of this salt measured in heavy water is shown in FIG.
In addition, using GL Science's Inertosyl ODS-2 column, 2.0 mM phosphate buffer (pH 6.6) was used as the eluent, and the high performance liquid chromatography was performed under the conditions of an outflow rate of 1.0 ml / min and detection at 210 nm. When carried out, the substance was eluted at a retention time of 5.0 minutes. When the prepared aqueous salt solution was examined by the DTNB method, the presence of free SH groups was not observed.

[0047]

Example 2 Production of 7-carboxy-3-{[(carboxymethyl) amino] carbonyl} -1-thia-4,6-diazaspiro [4.4] nonane (Production Method 1) Method described in Example 1 Glutathione-pyrrolidine sodium salt crude dry product 7.87g was manufactured in the same manner as in. This was extracted with 50 ml of methanol to obtain 5.70 g of a dried extract.

[0048] then dissolved to a total volume of 50% ethanol 368ml, QAE-SephadexA-25 HCO 3 at room temperature ^-
Column (3 × 31 cm) and ammonium bicarbonate in 50% ethanol was eluted with a linear gradient of 1,000 ml each of 0.08 M and 0.4 M solutions, collecting 17.8 ml fractions. It was Glutathione-pyrrolidine was eluted in the fractions 55 to 64. When this eluate was left at room temperature, the same behavior as the substance finally purified in Example 1 on high performance liquid chromatography was obtained. Shown generated.

When this material had grown to about 25% of the total (based on absorption at 210 nm), fractions 55 to 60 were combined and concentrated at 40 ° C. under reduced pressure to dryness. Dissolve the oily residue in 30 ml of methanol, then add 5 ml of ethanol,
Dried again. By repeating this dissolution and concentration operation, a white solid was obtained and dried over phosphorus pentoxide in a desiccator under reduced pressure for 3 days.

2.70 g of the dry solid thus obtained was dissolved in 93.5 ml of methanol, insoluble materials were removed by centrifugation, and 62 ml of ethyl acetate was added to the supernatant with stirring, and the resulting precipitate was centrifuged. Collected, dried similarly, 1.45g
To give a solid. 1.24 g of this solid was added to methanol 311
It was dissolved in ml, ethyl acetate (467 ml) was added with stirring, and the precipitate was dried to obtain 830 mg of a solid. This whole amount was again dissolved in 415 ml of methanol, 8.3 g of dithiothreitol was added, and the mixture was left standing overnight under a nitrogen stream, and then methylene chloride 1,0 was added.
After adding 37 ml and centrifuging at 12,000 xg for 30 minutes, the obtained precipitate was dried to obtain 444 mg of a solid.
417 mg of this solid was dissolved in 305 ml of methanol, 280 ml of methylene chloride was added with stirring, and the mixture was centrifuged in the same manner to remove the precipitate and obtain a supernatant. Methylene chloride 60 was added to this supernatant.
The precipitate formed when 0 ml was added was collected, dried and purified 7
-Carboxy-3-{[(carboxymethyl) amino]
Carbonyl} -1-thia-4,6-diazaspiro [4.
4] 197 mg of nonane was obtained (SIMS: 290 (M +
H) ⁺ ). Of this, 89 mg was converted to the sodium salt as in Example 1.

The ¹ H- ¹ H COZY NMR measured in heavy water is shown in FIG. This purified product is obtained in Example 1.
Under the same conditions as the high performance liquid chromatography described in
It was eluted at a retention time of 5.0 minutes. In addition, the presence of free SH groups was not recognized in the examination by the DTNB method.

[Brief description of drawings]

FIG. 1 is a diagram showing the synthesis of Example 1 according to a production method 2.
¹ H NMR of purified 7-carboxy-3-{[(carboxymethyl) amino] carbonyl} -1-thia-4,6-diazaspiro [4.4] nonane in heavy aqueous solution.
The spectrum is shown.

FIG. 2 is a diagram showing the synthesis of Example 2 according to the production method 1,
Purified 7-carboxy-3 - {[(carboxymethyl) amino] carbonyl} -1-thia-4,6-diazaspiro [4.4] ¹ H- ¹ H in a heavy water solution of nonane
The NMR spectrum by COZY is shown.

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Claims (6)

[Claims] 1. The formula [I]: [Wherein X represents H, COOH, R ¹ or COOR ¹ ,
Y represents OH, OR ¹ , A, AOR ¹ (here, R ¹
Each independently represents a C _{1 to} C ₈ linear, branched or cyclic hydrocarbon group, or a C _{1 to} C ₄ linear or branched hydrocarbon group substituted with an aromatic group, and A Represents an amino acid residue, and AOR ¹ represents an amino acid ester residue. )] Or a pharmaceutically acceptable salt thereof. 2. The group represented by X in the formula [I] is COOH, COOR ¹ (wherein R ¹ is C _{1 to} C _6).
Which represents a hydrocarbon group of), or a pharmaceutically acceptable salt thereof. 3. The group represented by Y in the formula [I] is OH, OR ¹ (wherein R ¹ represents a C _{1 to} C ₆ hydrocarbon group), Gly (wherein Gly represents a glycine residue). And Gly.OR ¹ (wherein Gly and R ¹ represent a glycine residue and a C _{1 to} C ₆ hydrocarbon group, respectively, which means a glycine ester residue as a whole). A compound or a pharmaceutically acceptable salt thereof. 4. The formula [II]: [Wherein X and Y are the same as in the case of the above formula [I]] to form a compound of the above formula [I] by subjecting the compound to a ring closure reaction in a weakly acidic, neutral or alkaline solution. And optionally obtaining a pharmaceutically acceptable salt of the compound of the formula [I], a process for producing the compound of the formula [I] or a pharmaceutically acceptable salt thereof. 5. The formula [III]: [Wherein X and Y are the same as in the case of the above formula [I]] to form a compound of the above formula [I] by subjecting the compound to a ring closure reaction in a weakly acidic, neutral or alkaline solution. And optionally obtaining a pharmaceutically acceptable salt of the compound of the above formula [I], a process for producing the compound of the above formula [I] or a pharmaceutically acceptable salt thereof. 6. Caused by limited replenishment of a compound of formula [I] comprising an effective amount of a compound of formula [I] or a pharmaceutically acceptable salt thereof. A preventive or therapeutic agent for diseases.