JPH0473425B2 - - Google Patents

Description

[Detailed description of the invention]

INDUSTRIAL APPLICATION FIELD OF THE INVENTION The present invention relates to glutamic acid and aspartic acid derivatives having antagonistic activity against biologically active polypeptides, and a method for producing the same. These derivatives are particularly useful as pain killers in the treatment of diseases of the digestive system and central nervous system, as well as anorexia and all diseases in which exogenous or endogenous biologically active polypeptides are involved, e.g. Tumors). Structure and Effects of the Invention The novel D,L-glutamic acid derivative and D,L-aspartic acid derivative according to the present invention are represented by the following general formula [], and also include pharmaceutically acceptable salts thereof. [In the formula, R and R′ are different from each other and are OH or R ₂ ,
n, R ₁ and R ₂ have the same meanings as described below.] That is, the compound of the above formula [] is specifically classified into the compounds of the following formulas [A] and [B]. [In the formula, n is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen, or cyano group or tri-substituted phenyl) (substituted with a fluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight chain, branched or cyclic alkyl group having 1 to 8 carbon atoms)] of the present invention It is recognized that the compounds to be protected have interesting pharmacological properties with respect to mammals. One of these properties is that it can enhance the analgesic activity of morphine and other analgesics. These properties are understood to be based, at least in part, on strong antagonistic activity toward cholecystokinin (CCK) or other biological regulatory peptides, which is exhibited by many of the compounds in question. be done. The compounds according to the invention can therefore be advantageously used in the treatment of various human diseases, such as diseases of the digestive system, for example in the treatment of colitis or biliary diskinesia, or in the treatment of etiology and severity. It can be used to treat pain. Furthermore, based on the pharmacological properties, the compound of the present invention has
The use of CCK or other biological polypeptides in the treatment of psychological disorders due to imbalances in physiological neuron levels, as well as in the treatment of anorexia, weight gain in agricultural animals, can be envisaged. Use in the treatment of diseases (eg, perhaps tumors) in which pathological cell development occurs through the promotion of or biologically active peptides is also envisaged. As mentioned above, the compound of the present invention has strong anti-inflammatory properties both in vitro and in vivo in various experimental models.
Has CCK activity. Accordingly, the compounds of the invention reduce CCK-induced contractions of the guinea pig gallbladder both in vitro and in vivo, inhibit induced contractions of the rabbit colon, and increase bile secretion in rats. It is also interesting that the compounds of the invention have an enhancing effect on the analgesic activity of analgesic anesthetics and analgesic non-anesthetics. This potentiating effect in fact makes it possible to considerably reduce the dose of anesthetic agent in the first stage, limiting a number of well-known undesirable side effects without significantly reducing the therapeutic index. The compounds of the invention can also be used to re-stabilize their analgesic activity when their pharmacological efficacy has decreased due to the well-known tolerance phenomenon in anesthetics, in which case the therapeutic dose can be increased. do not have to. Such advantageous therapeutic properties therefore also serve to gradually detoxify patients who have become addicted to the long-term use of anesthetics. Non-narcotic analgesics are extremely useful not only in their ability to increase analgesic activity but also in their ability to protect the gastric mucosa, which is generally damaged by such drugs. In particular, the activity-enhancing effect of analgesics is due to enkephalins, which have strong analgesic activity.
The ability of the compounds of the invention to block the hydrolysis of (endogenous physiological peptides) is relevant. This confers a greater half-life and greater activity than the enkephalins themselves. Pharmaceutical forms of the compounds of the invention can be prepared in a conventional manner, for example in tablets, capsules, suspensions, solutions and suppositories, and can be administered orally, parenterally or rectally. can.
The active ingredient is administered to the patient in an amount of, for example, 0.1 to 10 mg/kg body weight. For parenteral administration, it is preferred to use a water-soluble salt of the compound, such as the sodium salt or other non-toxic pharmaceutically acceptable salt. In addition, as inactive ingredients, substances commonly used for pharmaceutical purposes such as excipients, binders, fragrances, dispersants, colorants, and wetting agents are used. The method for producing glutamic acid derivatives and aspartic acid derivatives of the present invention is as follows: (a) Formula: [In the formula, n and R ₁ have the same meanings as above] An intramolecular anhydride represented by the formula: R ₂ H [In the formula, R ₂
has the same meaning as above], and 1 to 5
The above formulas [A] and [B] are obtained from the reaction solution at a molar ratio of -20°C to 30°C.
The method is characterized in that it includes a step of collecting the compounds and separating them. Note that the reaction temperature is preferably -10°C to 10°C. The intramolecular anhydride of the above formula [] is a novel compound that has never been produced before. Such intramolecular anhydride [ ] is obtained by (b) converting glutamic acid or aspartic acid under Schotten-Bauman conditions in an equimolar amount of the formula: R ₁ -CO-Cl (wherein R ₁ is as defined above). The formula: Then, (c) the compound [ ] is mixed with acetic anhydride in a molar ratio of 1 to 10 in a molar ratio of 1 to 10 in a compatible inert solvent.
It is produced by a process of reaction and dehydration at temperatures ranging from °C to reflux. The entire series of steps of the production method according to the present invention is described in the reaction steps below. The above acylation step b is carried out at a temperature of 0 to 15°C, 1 to
Preferably it is carried out under conditions of 24 hours, with a temperature of about 5° C. and a time of 12 hours recommended. In step c, the reaction time is, for example, about 30 minutes to 12
The time period is preferably about 3 hours, and the amount of acetic anhydride is preferably 3 mol per 1 mol of the compound []. In the amidation step a, the amine of formula: R ₂ H is preferably added in a molar ratio of 2.5 to 1 with the intramolecular anhydride [], and the reaction is carried out for about 30 minutes to 12 hours, preferably 3 hours. The relative proportions of compounds [A] and [B] are:
It varies depending on the type of R ₁ and R ₂ substituents used. Isomers A and B can be separated by fractional crystallization (using the solvents shown in Tables C and D below) or by extraction in basic media, but on average the more acid is compound [B] It is. Next, the present invention will be explained in more detail with reference to Examples. Example 1 Preparation of 3,4-dichloro-N-benzoylglutamic acid (compound A-4): - A solution of 14.7 g (0.1 mol) of L-glutamic acid in 200 ml of 1N sodium carbonate was cooled to 5°C,
To this, with stirring and cooling, 100 ml of 1N sodium carbonate and 21 g (0.1 mol) of 3,4-chlorobenzoyl chloride are added simultaneously over about 30 minutes. The mixture is left to react for 12 hours. This is acidified with concentrated HCl to a Congo red color and the precipitate that forms is filtered off. The residue is recrystallized from _H2O . Melting point 141-145℃, TLC
(See table below), Rf=0.46. Yield 24.5g (yield 76.4
%). All compounds of formula [] (see previous reaction steps) are prepared in the same manner as above. The resulting compounds are shown in Table A below (characteristic values for identifying them, yields, and solvents used for crystallization are also listed).

【table】

[Table] Example 2 Preparation of 3,4-dichlorobenzoylglutamic anhydride (compound B-4 in Table B): - 30.6 g (0.3 mol) acetic anhydride and 60 ml isopropyl ether to 32.0 g (0.1 mol) 3,
Add to 4-dichlorobenzoylglutamic acid.
The reaction solution is heated under reflux (73-77°C) for 2 hours. It is cooled, filtered, washed with a little ether to remove residual acetic anhydride, and dried. 27.0 g are thus obtained (yield 89.3%). Melting point 188~
190℃. All of the compounds of formula [] (see previous reaction steps) are prepared in the same manner as above. A number of specific examples of these compounds are shown in Table B below (characteristic values and yields for identifying them are also listed).

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[Table] Example 3 Preparation of D,L-4-(3,4-dichlorobenzoylamino)-5-(di-n-butylamino)-5-oxopentanoic acid (compound C-6 in Table C): - Charge 30.2 g (0.1 mol) of 3,4-dichlorobenzoylglutamic anhydride to the reactor and
ml of water. The reaction solution was cooled to about 5°C,
32.2 g (0.25 mol) of di-n-butylamino
Remove for several minutes. The mixture was left to react at this temperature for 3 hours,
Acidify with glacial acetic acid. This is filtered, washed with water until neutral, and dried. In this way 16.4g
(yield 38%). Melting point: 81-83°C (crystallized from isopropyl ether). TLC, Rf = 0.92. Expressions [A] and [B] in the same manner as above
(see previous reaction steps). A number of specific examples of these compounds are shown in Tables C and D below (characteristic values and yields identifying them are also listed).

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Table: The analgesic activity exhibited by the compounds protected by the present invention has been exemplified by a series of pharmacological tests demonstrating both the potentiating effect on the analgesic activity of anesthetic agents and the mechanism by which this potentiating effect is achieved. Ru. Experiment No. 1: Tail flick test (Tail
Increased analgesia of analgesic anesthetics in rats by Flick Test. Such a method is described by Harris, J. Pharmacol.
The method described in "Ther." ( 143 , pp. 141-148, 1964). Male rats weighing about 150-200 g without fasting are used. Select one point on the tip, irradiate it with a heat source (75°C), and measure the time (in seconds) that the rat stays still without moving the tip. A maximum time of 8 seconds under the heat source is chosen, after which the rats are removed in each case to avoid tissue damage. Measurements are taken before (control) and after drug treatment. Drug of the present invention (10mg/
Kg) was administered before the administration of morphine (2 mg/Kg).
Do it abdominally and vaginally for 10 minutes and just before. The rate of change for each rat is calculated using the following formula. Rate of change (%) = Time after treatment (seconds) - Administration time (seconds) / 8 - Administration time (seconds) x 100 Measurements were taken at 10, 20, 30, 45, 60 from the analgesic treatment.
and after 90 minutes. The results obtained are shown in Table 1. The table shows the treated group and dose, the mean rate of latent change in pain sensation (calculated for a group of 5 animals), the mean value (±SE) calculated from 1 to 90 minutes, and morphine alone or in combination with the compound of the invention. Record the administered potency ratio. From the data in Table 1, the test dose (10mg/Kgi.p.)
It has been found that most active products potentiate the activity of morphine, increasing its activity to about three times that of morphine alone.

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[Table] Experiment No. 2: Hot plate test (Hot-
plate test) Such a method is described by Edei et al. in J.Pharmac.
Exp. Ther.'' ( 107 , p. 385, 1953). A group of 5 non-fasted male rats weighing approximately 150 g are used. Metal plate at the bottom of a transparent cylindrical container [55± with azeotrope (acetone/ethyl formate = 1:1)
Heat to 1℃] and place the rat on top. The reaction time is the time interval from when the rat is placed on the hot plate until the rat licks its paw or attempts to jump out of the container. Control reaction times were 10 and 5 minutes before drug administration and 10, 20, 30, 45,
Measure after 60 and 90 minutes. Leave the rat on the plate for a maximum of 30 seconds. A response to product administration is considered positive if it is at least twice the normal reaction time. The results obtained are shown in Tables 2a and 2b. The table lists the treated group, dose and time spent on the plate (expressed as number of positive responses/number of treatments).
Record.

[Table] From the results in the table above, compound C-20 is 3mg/Kg
Even at ip doses, twice the analgesic activity of propoxyphene is observed. The actual maximum effect is obtained at a dose of 10 mg/Kgi.p.

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[Table] From the results in Table 2b, it can be seen that the compound of the present invention
mg/Kgi.p. is at least twice the analgesic activity of propoxyphen or methadone. In order to increase the analgesic activity of non-narcotic drugs, it is necessary to increase the dose of the drug of the present invention, and generally the larger the dose, the more significant the activity. Experiment No. 3: Effect of the compounds of the present invention on the analgesic activity of endogenous anesthetics released by transdermal ticks in rats (measured by tail flick test). .Neurosc.)” ( 1 , 358 pages, 1961)
This is the method described in . Male rats (weighing about 200 g) that are not fasted are used. Rats are stressed by applying a 60 Hz - 2.5 mA current to their paws with 1 second duration pulses every 5 seconds for 20 minutes. This stress regulation induces release of endogenous anesthetics. Immediately after electrical stimulation, rats were
Subject to tail flick test for the times indicated. The compound is given IV (intravenously) just before the electric shock.
(Dosages are shown in Table 3).

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[Table] From the data in Table 3, the compound of the present invention is 1 mg/Kg.
It has been observed that even doses of 100% can increase the analgesic activity of endogenous enkephalins to a very important degree. This increase is dose dependent, and the effect of this increase is dose dependent in nature, both in intensity and duration. Experiment No. 4: Enhancement of the analgesic activity of enkephalins induced by the compounds of the present invention In order to check one of the mechanisms of action of the compounds of the present invention, that is, the inhibitory effect on enzymatic degradation of endogenous enkephalins, the following procedure was performed. do an experiment. “Life Sciences” by Noble et al.
Male rats (used in groups of 5 rats) weighing 150-200 g were used to enable intracerebroventricular (icv) administration of the drug according to the method described in "Science" ( 6 , pp. 281-191, 1970). Insert the cannula into the right side of the chamber. Rats were then injected (icv) with the relevant compound at the doses shown in Table 4, and immediately after, 3 μg of D
- Treatment with ara-methionine-enkephalinamide (DALA). Analgesia is tested using the tail flick test at the times shown in Table 4. From the data in Table 4, it can be seen that the test component enkephalinamide (DALA) has an enhancing effect (both in intensity and duration) on the analgesic effect. This activity, which is extremely important even at a dose of 0.01 μg/Kg, is thought to be related to the inhibitory activity of enkephalins against enzyme(s) that responds to the metabolism of substances.

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[Table] Experiment No. 5: Anti-effect of the chemical of the present invention against the development of tolerance induced by repeated administration of morphine/HCl to rats. To measure ability;
The compounds of the present invention are tested. Weight approximately 200-250g
A group of 6 male rats are used. Each rat (excluding the physiologically treated control group) was given 10 doses of morphine hydrochloride ip at 5 mg/Kg at 24 hour intervals from the first treatment (time 0).
mg/Kgi.p. of the compound (except for the group treated with morphine alone). Pain thresholds are measured by tail flick test 15, 30, 45 and 60 minutes after treatment. The data shown in Table 5 is the average value of four measurements and shows the rate of change in latency time (pain appearance) before and after drug treatment. Also shown in Table 5 are Student's t values measured at various times comparing the drug treated group to the control group and the morphine treated group.

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[Table] Calculated regression line:
Group B: = activity = -0.327 x time + 48.41 (correlation coefficient = 0.95)
Group C: = activity = -0.118 x time + 45.08 (correlation coefficient = 0.76)
Group D: = activity = -0.320 x time + 67.97 (correlation coefficient = 0.93)
Group E: = activity = -0.251 x time + 66.47 (correlation coefficient = 0.89)
Group F: = activity = -0.314 x time + 70.38 (r = 0.9
6)
Group G: = activity = -0.245 x time + 66.06 (r = 0.7
9)
According to the data in Table 5 and the calculated regression line,
From the third treatment to the end of the experiment, groups C to G
It was observed that the activity was significantly greater than that of group B treated with morphine alone. Furthermore, after the fifth treatment, the morphine group differed significantly from the control group, while groups C-G
It is observed that superior activity is maintained compared to the control until the final treatment of 168 hours. Also, from the calculated regression line, the activity of the morphine group decreased to 0 on the 6th day of treatment, whereas
Groups C-G are seen to reach a level of inactivity between days 9 and 16. Next, the anti-CCK activity, anticonvulsant activity and choleretic activity of the compounds of the present invention will be illustrated. Anti-CCK activity in guinea pig gall in vitro Longitudinal sections of guinea pig gallbladder were incubated with oxygen/CO ₂ (95:5, V/
V) Temperature 32℃ while constantly oxidizing the mixture
Place in a diaphragm bath. Isometric contractions are detected and recorded with a force transducer. CCK-8 at a concentration of 10 μg/ml was used to contract the gallbladder, and the antagonistic activity of the compound of the present invention against the contractile effect of CCK was measured at different concentrations,
The ED50 value (ie, the concentration of compound capable of antagonizing 50% of the contractile effect of CCK, μg/ml) is determined. The results obtained are listed in the table below. The table shows the test compounds and the ED values (calculated by regression method from at least three experimental tests for each compound).

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[Table] From the data in Table 6, the compound of the present invention antagonizes CCK-8 activity by 50% at a certain concentration in the case of a highly active compound (for example, compound C-7), which is about 10 times that of a specific antagonist. , and exhibits excellent activity specificity. To corroborate the in vitro studies, some of the compounds of interest will be tested in vivo on guinea pig gallbladders in the field. Instructions for use are from Ljungberg's Svensk.Farm.Tidskr., 68 , pp. 351-354, 1964.
year). Guinea pigs weighing approximately 400 g are used, which are anesthetized with urethane. The test substance is injected into the jugular vein. The response of the gallbladder to the test substance is detected with a force transducer and a microdynamometer.
Record with . Optimal contraction dose of 10ng/Kg CCK-
Select as 8. The antagonist compound to be tested is administered in increasing doses to allow calculation of ED50, which is 50% of the contractile effect of CCK-8 at 10 ng/Kgi.v.
This is the dose (in mg/Kgi.v.) that can suppress the The results obtained are shown in Table 7. The table shows the usage amount and effect (expressed as inhibition rate of contraction effect of CCK-8).
Also state the ED50.

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[Table] These results essentially confirm what was found in the previous in vitro experiments. That is, the compound of the present invention is an extremely strong CCK antagonist, and the compound C-
Concentrations as low as 0.1 mg/Kg in the case of CCK-6 and C-7 can block gallbladder contraction induced by CCK-8 (even at concentrations clearly higher than physiological concentrations). In addition, the anticonvulsant activity exerted by the compounds of the present invention on the entire digestive system is also significantly observed. This activity was measured in a mouse vegetable carbon test (speed of gastrointestinal transit).
The results are shown in the table below.

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TABLE A more specific anticonvulsant activity more closely related to the physiological situation is illustrated in the following experiments. Make an incision in the abdomen of an anesthetized rabbit to reveal the transverse colon. A small ball filled with water is inserted into a fixed point and used as a pressure transducer.
Connect with a polyethylene cannula filled with water. The optimal sensitivity is fixed with respect to physiological conditions and the product is administered into the femoral vein. Administration of 100 ng/Kg CCK induces contractions. Table 9 shows the activity of the compounds of the present invention.

[Table] According to such data, the test compound of the present invention:
As previously shown in the case of the gallbladder, CCK is also shown to have an antagonistic effect on the intestinal contraction induced by administration at a high dose (100 ng/Kg). When using the best compounds, anticonvulsant activity is shown at very low doses of 1-3 mg/Kg. Another interesting feature of these compounds is that they significantly increase bile flow rate. Perform the experiment shown below. A cannula is inserted into the bile duct of a rat anesthetized with urethane, together with a small needle connected to a polyethylene tube, and bile is collected. Collections are made 1 hour before and 2 hours after intravenous administration of test compound, and collected samples are weighed at 30 minute intervals. To prevent dehydration of the rats, administer 0.5 ml of physiological solution every 30 minutes for a total of no more than 3 ml. The results obtained for some of the compounds of the invention are shown in the following table. It is expressed as ED50, which is the amount of substance in ml/Kgi.v. that can cause a 50% increase in bile circulation after drug treatment compared to the control value (average value measured during one hour of collection before drug treatment). (average value measured over 2 hours). From such data, it is inferred that the compound has a strong choleretic activity. The average ED50 of the test compounds is 5-25 mg/Kg, with remarkable agreement between dose and pharmacological response (actual correlation coefficients are greater than 0.90 in all cases).

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[Table] The anti-inflammatory properties exhibited by most of the compounds
To check the hypothesis that CCK activity could be advantageously used in the treatment of anorexia in humans or as an appetite stimulant in agricultural animals, the following experiments are performed. Male rats weighing approximately 160 g divided into groups of 10 are used. For each group for 3 weeks,
Give the indicated dose of drug daily. The drug in the form of sodium salt is dissolved in water,
A volume of 10 ml/Kg of H ₂ O is administered, while the control group receives the same volume of water only. Average feed consumption and average body weight for each group, calculated weekly, and Student's t calculated from various treatment and control groups.
The values are shown in the table below. From the data in Tables 11 and 12, the daily dose of 0.3mg/
Kg of Compound C-7 reduced feed consumption by approximately
A 15% increase is permitted. This increase is approximately 30% in other dose studies and is always extremely significant. The weight gain of treated rats follows a similar course compared to that of control rats. During the study period, all groups treated with C-7 produced significantly greater weight gain than control rats.

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[Table] Inhibitory effect on the growth of pancreatic adenocarcinoma induced by CCK-8 The compound of the present invention, which has the most potent anticholecystokinin effect on the trophic activity of CCK in normal pancreatic cells and pancreatic adenocarcinoma, That is, compound C-7 will be studied. 1×10 ⁵ pancreatic adenocarcinoma in the cheek sac of a male hamster
Inoculate a suspension of tumor cells. Five days after inoculation, hamsters are randomly divided into four groups of 10. i.e. control group, 10μg/Kg three times a day
Group treated with CCK-8, 3 times a day 5
a group treated with compound C-7 at mg/Kgi.p.
and compounds C-7 and CCK-8, respectively, in the same manner as above. After 15 days of treatment, the hamsters are sacrificed, the normal pancreas and the inoculated pancreatic tumor in the cheek pouch are collected and weighed. Extract the DNA and measure using standard methods. The results obtained are shown in Table 13. Mean value (±S.
Displayed in E.).

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[Table] According to the data in Table 13, cholecystokinin hormone (CCK-8 hormone is a physiologically active component), which has trophic activity on normal pancreatic cells, stimulates the growth of pancreatic cancer. I understand. Compound C-7, a potent specialized CCK antagonist
very significantly antagonizes these effects of CCK-8. According to the above experimental data, the use of compound C-7 or other anti-cholecystokinin compounds protected by the present invention is sustained by endogenous biologically active polypeptides, in particular CCK. tumor,
It is believed that this will lead to particularly favorable results in the treatment of, for example, gastrointestinal and pancreatic tumors. Such experimental data also indicate that the use of the agents of the present invention in conjunction with morphine or other analgesics (both narcotic and non-narcotic) represents a significant therapeutic innovation, relieving the etiological pain. This suggests that we may be able to provide compounds that are of great interest to physicians.
This treatment may be especially necessary in the case of long-term administration of anesthetics, in which case the drug may become habit-forming or
Or at least it is extremely necessary to keep it within acceptable limits. Moreover, their use in the detoxification of patients who have become dependent on long-term use of anesthetics is likely to be of extraordinary therapeutic community interest. The above experimental data also confirm that these compounds are useful in the treatment of various pathological conditions of the gastrointestinal system, such as in the treatment of spasmodic syndromes in general and pain relief and in particular in the treatment of biliary insufficiency and irritable colon. Based on the above, we conclude that the compounds of the present invention exhibit potent anti-CCK activity, which is demonstrated by many of them, in the treatment of anorexia, or in the treatment of CCK at physiological neuron levels.
Alternatively, suitable therapeutic uses of other biologically active peptides in the treatment of pathological symptoms of SNC related to imbalance may be identified.

Claims (1)

[Claims] 1 formula, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or substituted amino (substituted with the same or different straight chain, branched or cyclic alkyl group having 1 to 8 carbon atoms) ] A pharmaceutically active D,L-glutamic acid derivative or a D,L-aspartic acid derivative, or a pharmaceutically acceptable salt thereof. 2 R is OH, R' is R ₂ , n is 2, R ₁ is 3,4-dimethylphenyl or 3,4-dichlorophenyl, and R ₂ is a C ₄ to _{C 5} linear alkyl group The glutamic acid derivative according to item 1 above, which is a substituted amino acid or a pharmaceutically acceptable salt thereof. 3. The compound according to item 1 above, wherein R is R ₂ , R' is OH, n is 2, R ₁ is 4-cyanophenyl, and R ₂ is amino di-substituted with a C ₄ to C ₅ linear alkyl group. A glutamic acid derivative or a pharmaceutically acceptable salt thereof. 4 Formula as active ingredient, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms); Biologically active polypeptides characterized by comprising a compound represented by [1] or a pharmaceutically acceptable salt thereof, particularly for the treatment of SNC diseases related to imbalance at the physiological neuron level of cholecystokinin. Therapeutic pharmaceutical composition. 5 Formula as active ingredient, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms); A pharmaceutical composition for use as an anticonvulsant, characterized by comprising a compound represented by the following formula or a pharmaceutically acceptable salt thereof. 6 Formula as active ingredient, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms); A pharmaceutical composition for use as a choleretic agent, characterized by comprising a compound represented by the following or a pharmaceutically acceptable salt thereof. 7 Formula as active ingredient, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms); A pharmaceutical composition for treating anorexia, comprising a compound represented by the following or a pharmaceutically acceptable salt thereof. 8 Formula as active ingredient, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms); A pharmaceutical composition for the treatment of tumors involving cholecystokinin or biologically active polypeptides having a similar mechanism of action, characterized by comprising a compound represented by the following formula or a pharmaceutically acceptable salt thereof: thing. 9 Formula as active ingredient, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms); A pharmaceutical composition for use as a pain suppressant for humans, which comprises a compound represented by the following formula or a pharmaceutically acceptable salt thereof, and is used in combination with an analgesic. 10 To increase the rate of weight gain of agricultural animals,
It is used as an appetite stimulant in animals, and as an active ingredient it contains the formula, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with the same or different straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms); A preparation for animal breeding containing at least one compound represented by: 11 formula, [In the formula, R and R′ are different from each other and are OH or R ₂ , n
is 1 or 2, R ₁ is mono-, di- or tri-substituted phenyl (substituted with the same or different linear or branched C ₁ -C ₄ alkyl group, substituted with halogen,
or substituted with a cyano group or a trifluoromethyl group), and R ₂ is morpholino, piperidino or mono- or di-substituted amino (substituted with a straight-chain, branched or cyclic alkyl group having 1 to 8 carbon atoms). A method for producing a D,L-glutamic acid derivative or a D,L-aspartic acid derivative shown, or a pharmaceutically acceptable salt thereof, comprising (a) the formula: [In the formula, n and R ₁ have the same meanings as above] An intramolecular anhydride represented by the formula: R ₂ H [In the formula, R ₂
is the same meaning as above] and 1 to 5
The reaction was carried out at a molar ratio of
A manufacturing method comprising the steps of recovering and separating a compound in which R' is R ₂ and R is R ₂ and R' is OH. 12 The intramolecular anhydride of the formula [ ] used in step (a) of Section 11 above, (b) glutamic acid or aspartic acid under Schotten-Bauman conditions in an equimolar amount of the formula: R ₁
-CO-Cl is reacted with acyl chloride at a temperature of -20°C to 30°C, formula: (c) The compound [] is treated as such or with acetic anhydride in a molar ratio of 1 to 10 in a compatible inert solvent.
12. The method according to the above item 11, which is obtained by a step of reacting and dehydrating at a temperature of .degree. C. to reflux. 13 n is 2, R ₁ is 3,4-dimethylphenyl or 3,4-dichlorophenyl, and R ₂ is
13. The method according to the above item 11 or 12, wherein the amino acid is di-substituted with a _C4 to _C5 straight chain alkyl group. 14. The method according to item 11 or 12, wherein 14n is 2, _R1 is 4-cyanophenyl, and _R2 is amino di-substituted with a _C4 - _C5 linear alkyl group.