JPH0533716B2 - - Google Patents

Description

Detailed Description of the Invention <Industrial Field of Application> The present invention relates to a 4-oxo-2-azetidinecarboxylic acid derivative represented by the following general formula [] and a salt thereof, which is useful as a medicine. (In the formula, R ¹ is an azide group, an amino group, a lower acylamino group, a mercapto group, or a lower alkylthio group, and R ² is the formula [Formula] or [Formula]) In the formula, R ³ is a hydrogen atom or a lower means an alkyl group. An imidazolyl group represented by 〓, Y means a hydroxyl group, a lower alkoxy group, an amino group, or a mono- or di-lower alkylamino group.) <Specific Description of the Invention> The object compound of the present invention [ ] will be further explained as follows. "Lower" in "lower acylamino group" and "lower alkylthio group" meant by R ¹ and "lower alkyl group" meant by R ³ has 1 to 5 carbon atoms.
straight or branched carbon chain. Therefore, lower acylamino groups include acetylamino groups, propionylamino groups, butyrylamino groups,
Examples of lower alkylthio groups include methylthio group, ethylthio group, propylthio group, isopropylthio group, butylthio group, and pentylthio group. group, ethyl group, propyl group, butyl group, isopropyl group, sec-butyl group, etc. The lower alkoxy group represented by Y is a straight chain or branched group having 1 to 5 carbon atoms such as methoxy group, ethoxy group, propoxy group, isopropoxy group, butoxy group, tert-butoxy group, pentyloxy group, etc. It is a lower alkoxy group. The object compound of the present invention [ ] has at least three asymmetric carbon atoms, and stereoisomers based on this exist. The target compounds of the present invention include separated and mixtures of these isomers. Further, the object compound of the present invention [] forms a salt with an acid or a base. Salts encompassed by this invention include salts with non-toxic acids (e.g., inorganic acid salts such as hydrochlorides, sulfates, and organic hydrochloric acids such as citrates, acetates, tartrates); Examples include salts with bases (for example, salts with inorganic bases such as sodium salts and potassium salts, and salts with organic bases such as ammonium salts and trimethylamine salts). The object compound of the present invention [] can be prepared as it is or mixed with appropriate pharmacologically acceptable carriers, excipients, and diluents to form powders, granules, tablets, capsules, etc.
It can be administered orally or parenterally in the form of injections (intravenously, subcutaneously, intramuscularly), suppositories, and the like. <Effects and uses of the invention> The dosage of the compound [ ] of the present invention varies depending on the type of compound [ ], age, body weight, symptoms, route of administration, etc., but for example, in the case of injection, approximately
0.001-10mg preferably 0.01-0.1mg (single dose)
and about 0.05 to 500 mg for oral administration, preferably 0.1
~10 mg (single dose). Compounds related to the object compound [ ] of the present invention are also known as "thyrotropin-releasing hormone".
(TRH) L-pyroglutamyl-L-histidyl-L-prolinamide (pGlu-His-Pro
−NH ₂ ) is known. The existence of TRH has been known since the 1960s, but its structure was confirmed in 1970 [Endocrinology, 86 , 1143 (1970)]. TRH is
It was originally thought to be a hormone that regulates the release of thyrotropin (TSH) in the pituitary gland of mammals. However, subsequent studies have shown that the biological functions of this tripeptide TRH are not limited to the regulation of TSH release, but also to the central nervous system (CNS).
This discovery opened a new field of research [Science,
178, 417 (1972), Lancet, 2 , 999 (1972)]. Thus, in addition to its TSH-releasing activity, TRH also plays a role in reducing sleep duration induced by barbiturates and alcohol, suppressing hypothermia symptoms stimulated by various drugs, increasing locomotor activity, and inhibiting haloperidol-induced sleep duration. It is known to have effects on the CNS, such as preventing generalized tonicity caused by this disease, enhancing memory, improving symptoms of schizophrenia, and antidepressant effects. Furthermore, TRH has been found to be an ameliorating and therapeutic agent for disorders of consciousness caused by functional or organic disorders in the brain, such as head trauma, brain surgery, cerebrovascular disorders, brain tumors, etc., especially acute or subacute disorders of consciousness. (Unexamined Japanese Patent Publication No. 51-118841)
issue). In terms of clinical treatment, TSH release activity is weaker than that of TRH, or shows almost no activity, and moreover, as mentioned above,
There has been a demand for a TRH derivative that has an effect on the CNS equal to or greater than that of TRH. Various TRH derivatives have been synthesized for such purposes, and their effects on the CNS have also been further expanded. Compounds synthesized for this purpose include, for example, their TSH release activity is weaker than that of TRH, but they have anesthetic antagonism, increase locomotor activity, or dopamine-like effects, leading to sleep drug addiction, consciousness disorders, hyperactive children, and schizophrenia in humans. , TRH derivatives that are said to be useful for improving treatment of depression and Parkinson's disease (Japanese Patent Publication No.
No. 52-116465), has an effect on consciousness disorder after head trauma, and has an effect on reducing sleep duration due to hexobarbital, and is effective for patients with consciousness disorder caused by organic or functional disorders in the brain, aging or mental fatigue. TRH derivatives (Japanese Patent Application Laid-Open No. 1983-1989) are said to be useful for treating patients with symptoms such as depression and depression.
No. 59714) is known. The object compound of the present invention is a TRH derivative in which the pyroglutamyl (pGlu) structural part of TRH is converted into a 4-oxo-2-azetidinyl carbonyl structure (β-lactam structure), which has not been used at all in the past. It has chemical structural characteristics, and in terms of pharmacological action, it has a significantly stronger ONS action than the above-mentioned TRH and conventionally known TRH derivatives, and is useful as a medicine. For example, schizophrenia, depression,
An agent for improving consciousness disorders or loss of motivation in cerebrovascular accident sequelae, head trauma, senile dementia, epilepsy, etc.
It is useful as an ameliorating agent for depression, memory loss, etc. The pharmacological effects of the compounds of the present invention were measured as follows. The results are shown together with the measurement method. Experimental Example 1 (Effect on hypothermia caused by pentobarbital) Tests are carried out on groups of 9 male mice each weighing between 18 g and 22 g for each dose of test compound. Pentobarbital sodium salt 55mg/Kg
is administered intraperitoneally, and 10 minutes after administration, TRH or the test compound is administered intravenously to the animal. The rectal temperature of the animals was recorded 30 minutes after administration of the test compound, and the results were compared with a control group that received intravenous saline alone.
Expressed as the dose that increases rectal temperature by 1.5°C (ED 1.5°C). The results are shown in Table 1. [Table] <Production method> According to the present invention, the target compound is produced by the following route. (In the formula, R ¹ , R ² , R ³ and Y have the above-mentioned meanings. Y' means a hydroxyl group, an amino group, or a mono- or di-lower alkylamino group.) That is, according to the present invention, the target compound [] is (a) reacting a compound [] with a compound [] to make a compound [], and then this compound []
and the compound [], or (b) reacting the compound [] with the compound [] to form the compound [], and then reacting the obtained compound [] with the compound []. It can be manufactured by folding. Further, the thus obtained target compound [ ₁ ] can also be led to another target compound [ ₂ ] by converting the substituent Y. Compound employed in (a) or (b) above [ ₁ ]
The production reaction is a peptide synthesis reaction, and a method known per se is used. Commonly used methods include a method using dicyclohexylcarbodiimide as a condensing agent, an azide method, an acid chloride method, an acid anhydride method, and an active ester method. To carry out these methods, prior to the peptide formation reaction in each step, functional groups such as amino groups, imino groups, carboxy groups, etc., which do not participate in the reaction of the raw material compound are protected, and The amino group, imino group or carboxyl group involved in the peptide formation reaction is activated if necessary. A compound in which an amino group, an imino group, or a carboxyl group is activated, such as an active ester, may be once isolated and then subjected to a peptide synthesis reaction, or may be subjected to a peptide synthesis reaction without being isolated. Examples of protecting groups for amino groups include benzyloxycarbonyl group, t-butyloxycarbonyl group, p-methoxybenzyloxycarbonyl group, phthaloyl group, and trifluoroacetyl group. Examples of protecting groups for imino groups include, for example. tosyl group, benzyloxycarbonyl group, p-methoxybenzyloxycarbonyl group, benzyl group,
Examples include 2,4-dinitrophenyl group. As protective groups for carboxyl groups, methyl ester, ethyl ester, benzyl ester, p-
Ester forms such as nitrobenzyl ester and t-butyl ester are used. The groups involved in the reaction can be activated by using the phosphazo method using phosphorus trichloride, the isocyanate method using phosgene, or the phosphite method when the group is an amino group or an imino group, or the activation of a carboxyl group. When , active ester (2,4-dinitrophenol ester, N
-hydroxysuccinimide ester, etc.), azide, carboxylic acid anhydride, etc. Among these, the azide method or the method using dicyclohexylcarbodiimide as a condensing agent is preferred for synthesizing compounds [] and []. It is also possible to use a method of directly preparing a peptide using an N-carboxylic anhydride of an amino acid without using a protecting group. Next, the peptide-forming reaction is carried out in a conventional manner in an inert solvent by cooling or heating. Suitable solvents include dimethylformamide (DMF), ethyl acetate, dichloromethane (methylene chloride), and tetrahydrofuran. When it is necessary to remove a protecting group from a reaction product, for example, in the case of a benzyl ester, by catalytic reduction, and in the case of a p-toluenesulfonyl group as a protecting group for an imino group, anhydrous hydrogen fluoride,
By using HOBT or hydrogen fluoride-pyridine complex, etc., when the protecting group is an alkyl ester, it is hydrolyzed, and when the protecting group is benzyloxycarboxinyl or p-methoxybenzyloxycarbonyl, it is catalytic reduction or hydrogen bromide. By acid-acetic acid treatment, when the protecting group is a t-butyloxycarbonyl group, it can be easily removed by acid decomposition. Next, for the reaction of converting the substituent Y of the target compound [ ] of the present invention to lead to another target compound, appropriate reaction conditions are adopted depending on the properties of the compounds involved in the reaction. <Examples> The present invention will be further described below with reference to Examples. In addition, the manufacturing method of the raw material compound used in an Example is shown in a reference example. The abbreviations used in Examples and Reference Examples have the following meanings. NMR Nuclear magnetic resonance spectrum IR Infrared absorption spectrum Mass Mass spectrometry spectrum mp Melting point His Histidine Pro Proline DMF Dimethylformamide HOBT 1-Hydroxy-1,2,3
-Benzotriazole TEF Tetrahydrofuran DCC Dicyclohexylcarbodiimide Reference Example 1 (Raw material of Example 1) Dry 1 to 3.4 g of (S)-1-t-butyldimethylsilyl-4-oxo-2-azetidinecarboxylic acid.
It is dissolved in 30 ml of THF and added to a solution of 30.6 mmol of lithium diisopropylamide in 22 ml of dry THF at 0°C. After removing the cooling bath and stirring for 35 minutes, the mixture was cooled to -70°C, and a solution of 3.5 g of p-toluenesulfonylazide dissolved in 18 ml of THF was added dropwise. This is stirred at -50°C for 1 hour and cooled again to -70°C. After adding 4.81g of trimethylsilyl chloride
Warm to 40°C and stir for 6 hours. The reaction mixture was concentrated under reduced pressure, and 3.7 g of sodium hydrogen carbonate was added to the residue.
Add a solution dissolved in 100 ml of water, and adjust the pH to 3 with a 10% citric acid aqueous solution. This aqueous solution is extracted three times with ether, and the ether layers are combined and dried over anhydrous sodium sulfate. After filtration, concentrate the solution in 3.2
g of oil is obtained. Add this to silica gel (100g)
(2S,3R)-1-t-butyldimethylsilyl-3-azido-4 was obtained by column chromatography and elution with n-hexane-ethyl acetate (4:1) and (2:1). 2 to 1.6 g of -oxo-2-azetidinecarboxylic acid are obtained as a colorless solid. (i) NMR (CDCl ₃ ) δppm: 0.18 (3H, s,
CH ₃ ), 0.32 (3H, s, CH ₃ ), 0.98 (9H, s,
t _Bu ), 4.00 (1H, d, J=3Hz), 4.71 (1H, d,
J=3Hz) (ii) IR (KBr) cm ^-1 : 2095, 1740, 1700 Dissolve 1.78 g of compound 2 in 39 ml of methanol,
Add 9.85 ml of 1N hydrochloric acid to this and stir at room temperature for 1.75 hours. The reaction mixture was cooled to 0°C, 9.85 ml of 1N sodium hydroxide was added, and the mixture was concentrated to dryness under reduced pressure to obtain an oil. Dissolve this in 10ml of DMF and
After adding a molecular sieve and allowing it to stand overnight, the solvent was distilled off under reduced pressure and the (2S,3R)-3-azide
4-oxo-2-azetidinecarboxylic acid 3- was obtained. This compound was used as it was in the following Example 1. Example 1 Compound 3~ obtained in Reference Example 1(b) was dissolved in a mixed solvent of 15 ml of DMF and 15 ml of methylene chloride HOBT1.06
Add g and cool on ice. After adding 1.62 g of DCC to this, the mixture was stirred at 0°C for 40 minutes. (Reaction solution A) On the other hand, L-histidyl-L-prolinamide 2
Dissolve 4 to 2.71 g of hydrobromide in 32 ml of DMF, -
Add 2 ml of triethylamine at 15°C. This-
The mixture was stirred at 5° C. for 20 minutes to remove the precipitated salt, yielding a DMF solution of the free base (reaction solution B). Add reaction solution B to reaction solution A and stir at 0 to 5°C for 20 hours. After removing the insoluble materials, the liquid was concentrated under reduced pressure and the resulting oil was subjected to column chromatography on silica gel (180 g). When eluted with chloroform-methanol-aqueous ammonia (80:20:2), N〓[(2S,3R)-3-
Azido-4-oxo-2-azetidinylcarbonyl]-L-histidyl-L-prolinamide 5~ was obtained and lyophilized to give 58 mg as a colorless solid. (i) NMR (d ⁶ -DMSO) δppm: 1.86 (4H, m),
2.85~3.00 (2H, m) 4.08 (1H, d, J=2
Hz), 4.6-4.8 (2H, m), 6.96 (1H, s), 7.00
(1H) 7.58 (1H, s), 8.06 (1H), 8.65 (1H,
d, J = 8Hz), 8.78 (1H) (ii) IR (KBr) cm ^-1 : 3400, 2110, 1765, 1620~
1680 (Broad) (iii) Mass (FAB) m/z: 390 (M+1), 362,
307 Reference example 2 (raw material of Example 2) Compounds 1 to 920 mg are dissolved in 8 ml of dry THF and added to a solution of 8.28 mmol of lithium diisopropylamide in 6 ml of THF at 0°C. Then, the cooling bath was removed, the mixture was stirred for 35 minutes, and then cooled to -70°C. Add 0.47ml of dimethyl disulfide to -70 to -60
Stir for 30 minutes at -50°C for 1 hour. Remove the cooling bath, continue stirring, and when the temperature reaches 0℃, add 5 ml of 10% citric acid aqueous solution, 30 ml of ice water, and ether.
Pour into 50ml of the mixture. with 10% citric acid aqueous solution
Adjust the pH to 3-4 and separate the aqueous layer and organic layer. The aqueous layer is extracted with 30 ml of ether, and the organic layers are combined and washed with water. After drying over anhydrous magnesium sulfate, the solvent was removed under reduced pressure to obtain a pale yellow oil. This is purified by column chromatography on silica gel (80 g). Elution with n-hexane-ethyl acetate (4:1) gave 6 to 272 mg of (2R,3R)-1-t-butyldimethylsilyl-3-methylthio-4-oxo-2-azetidinecarboxylic acid as a white solid. Ta. (i) NMR (CDCl ₃ ) δppm: 0.16 (3H, s,
CH ₃ ), 0.32 (3H, s, CH ₃ ), 0.97 (9H, s),
2.19 (3H, s, SCH ₃ ), 4.00 (1H, d, J=3
Hz), 4.27 (1H, d, J = 3Hz) Dissolve 6 to 569 mg of compound in 12 ml of methanol,
Add 3 ml of 1N hydrochloric acid and stir at room temperature for 1.75 hours. The reaction solution was cooled to 0°C, and 1N-sodium hydroxide 3
ml and concentrated to dryness under reduced pressure. This was freeze-dried as an aqueous solution to obtain a white powder containing (2R,3R)-3-methylthio-4-oxo-2-azetidinecarboxylic acid 7~. This was used as a raw material for the next example without purification. Example 2 The white powder containing 7~ obtained in Reference Example 2(b) was
Dissolve in a mixture of 5 ml of DMF and 5 ml of methylene chloride,
Add 324mg of HOBT, then 618mg of DCC under ice cooling.
Add and stir for 40 minutes. A DMF solution of 2 mmol of L-histidyl-L-prolinamide (free base) prepared as in Example 1 is added to the reaction mixture. After stirring overnight at 0-5°C, insoluble matter is removed. The liquid was concentrated under reduced pressure and the resulting oil was subjected to column chromatography on silica gel (100 g). When eluted with a mixture of chloroform-methanol-ammonia water (80:20:2), N〓 [(2R, 3R)
-3-Methylthio-4-oxo-2-azetidinylcarbonyl]-L-histidyl-L-prolinamide 8~ was obtained as a colorless solid after lyophilization.
Get 236mg. (i) mp142-144℃ (ii) NMR ( ^d6 -DMSO) δppm: 1.85 (4H, m),
2.06 (3H, s, SCH ₃ ), 2.84-2.96 (2H, m),
6.93 (1H, s), 6.98 (1H), 7.56 (1H, s) 8.04
(1H), 8.5 (2H, m). (iii) IR (KB _r ) cm ^-1 : 3400 (broad), 1755,
1620-1680 (broad) (iv) Mass (FABm/z: 395 (M+1), 381, 349 Reference example 3 (raw material of Example 3) (±) 9 to 1.67 g of cis-3-azido-4-oxo-2-azetidinecarboxylic acid methyl ester,
1.86g of paratoluenesulfonic acid monohydrate in DMF50
ml, add 400 mg of 10% Pd-C, and hydrogenate at room temperature and pressure. The catalyst is removed and the solvent is distilled off under reduced pressure to give a viscous oil. This was dissolved in 80 ml of methylene chloride, cooled to -10°C, and 2.89 ml of triethylamine and 0.736 ml of acetyl chloride were sequentially added.
Stir at the same temperature for 1 hour and 20 minutes. The reaction mixture was concentrated under reduced pressure and the residue was subjected to column chromatography on silica gel (100 g). (±) by elution with ethyl acetate-methanol (10:1)
Cis-3-acetylamino-4-oxo-2-azetidinecarboxylic acid methyl ester 11-683mg
is obtained as a white solid. (i) NMR (CDCl ₃ + CD ₃ OD) δppm: 1.98 (3H,
s, CH ₃ CO) 3.77 (3H, s, OCH ₃ ), 4.46
(1H, d, J=6Hz), 5.45 (1H, d, J=6
Hz) (ii) IR (KBr) cm ^-1 : 3250, 3175, 3080, 1745,
1650 Compound 11 ~ 497 mg was dissolved in 40 ml of methanol,
Add 2.7 ml of sodium hydroxide under ice cooling, stir for 25 minutes under ice cooling, then remove the cooling bath and stir for 30 minutes. Cool on ice again, add 2.7 ml of IN hydrochloric acid, and concentrate to dryness under reduced pressure. The residue was azeotropically dehydrated under reduced pressure using an acetonitrile-benzene mixture to obtain a residue containing (±)cis-3-acetylamino-4-oxo-2-azetidinecarboxylic acid 12~. This was used as it was in the next example. Example 3 Example 2 The residue containing compounds 12~ obtained in Reference Example 3(b) and L-histidyl-L-prolinamide (free base) prepared in the same manner as in Example 1 were added.
React using the same procedure as above. The resulting reaction mixture was purified by column chromatography on silica gel (150 g). Ethyl acetate-methanol-aqueous ammonia (20:
10:1) to elute the target compound, remove the solvent, and freeze-dry. N〓[(2S,3S)-3-acetylamino-4-oxo-2-azetidinylcarbonyl]-L-histidyl-L-prolinamide 13a~ and N〓-[(2R,3R)
274 mg of a mixture of -3-acetylamino-4-oxo-2-azetidinylcarbonyl]-L-histidyl-L-prolinamide 13b is obtained. (i) NMR (D ₂ O) δppm: 1.88-2.10 (7H), 2.95
~3.16 (2H), 3.5 (1H, m), 3.8 (1H, m), 4.4
(1H, m), 4.57 (1H, m), 4.7~5.04 (1H,
m), 5.35 (1H, m), 7.08 (1H), 7.88 (1H), (ii) IR (KBr) cm ^-1 : 3350 (broad), 1755,
1620-1680 (Broad) Mass (FAB) m/z: 406 (M+1), 363,
235

Claims (1)

[Claims] 1. General formula (In the formula, R ¹ is an azide group, an amino group, a lower acylamino group, a mercapto group, or a lower alkylthio group, and R ² is the formula [Formula] or [Formula]) In the formula, R ³ is a hydrogen atom or a lower means an alkyl group. 4-oxo-2-azetidinecarboxylic acid represented by an imidazolyl group represented by 〓, Y means a hydroxyl group, lower alkoxy group, amino group, or mono- or di-lower alkylamino group) Derivatives and their salts.