JPS6281385A - Novel methotrexate derivative - Google Patents

Abstract

NEW MATERIAL:A compound shown by the formula I (A<1> and A<2> are H or ester residue; R is 1-5C alkyl; 1-5C alkylthio or OH) or its salt. EXAMPLE:N-[4[[2,4-Diamino-6-pteridinyl)methyl]methylamine]benzoyl]7-meth ylthioglutamic acid shown by the formula IV. USE:An antimalignant tumor agent. Suitable for a large amount of administration, having low toxicity. PREPARATION:A raw material substance shown by the formula II is reacted with a 2,4-diamino-S-halomethylpteridine shown by the formula III in a solvent such as DMF, etc., at 15-120 deg.C, to give an ester derivative shown by the formula Ia. Then, the ester derivative is treated with water and an inorganic base (e.g., NaOH, etc.,) in a solvent such as methanol, etc., preferably at 15-25 deg.C, to give a compound shown by the formula Ib.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel methotrexate derivatives useful as anti-malignant tumor agents.

BACKGROUND OF THE INVENTION Methotrexate (US Pat. No. 4,057,578) has been used to treat various malignant tumors such as leukemia and trophoblastic tumors. The present inventors have also conducted extensive research on methotrexate derivatives and have discovered a group of compounds described in Japanese Patent Application No. 166911/1989 filed on August 8, 1980.

Problems to be resolved Since methotrexate is highly toxic, there has been a desire to develop an anti-malignant tumor agent with low toxicity and suitable for large-dose administration.

The present inventors have discovered that a novel methotrexate derivative represented by the following general formula (1) or a salt thereof has an excellent anti-malignant tumor effect, and has developed the present invention. completed.

General formula: (In the formula, A1 and A2 each represent hydrogen or an ester residue; R represents 01-C6 alkyl, Cl-06 alkylthio, or hydroxy.) The above definition will be explained below.

C, -C, alkyl and 01-C, alkyl in C□~C, alkylthio include methyl, ethyl, n
-propyl, isopropyl, n-butyl, isobutyl,
5ee-butyl, tart-butyl, n-pentyl, isopentyl, 5ee-pentyl, neo-pentyl,
Examples include tert-pentyl.

Examples of ester residues include, in addition to the above-mentioned C1-C, alkyl, phenyl which may have a substituent (e.g., nitrophenyl, methoxyphenyl), phenyl CI'' Cs alkyl (C, -C, Cl) ), etc.

The compound of the present invention (I> is produced by production method A and/or production method B)
It can be manufactured by

(Left below) Manufacturing method A (wherein, R has the same meaning as above, A to and A"
each represents an ester residue, and )lal represents a halogen (chlorine, bromine, iodine, etc.). ) The above manufacturing method will be explained in detail below.

The reaction between the first step raw material (I[) and 2,4-diamino-6-barometerbuteridine (I[) is carried out in a suitable solvent for 15 to 1
It may be carried out at a temperature of 20°C. Examples of the solvent include dimethylformamide, dimethylacetamide, dimethylsulfoxide, chloroform, hexamethylphosphoric triamide, and a mixed solvent thereof. A suitable base (eg, triethylamine, pyridine, dimethylaniline, etc.) may be added to accelerate the reaction.゛ (Hereinafter, blank space) Second step Next, the hydrolysis of the ester (Ia) is carried out by a conventional method, but in general, water and an inorganic base (for example, alkali hydroxide, alkali hydrogen carbonate, carbonate) are used in a suitable solvent. alkaline) and 15-100°C, preferably room temperature (15-25°
C) can be used.

Examples of the solvent include alcohols such as methanol, ethanol, and isopropanol, solvents that are miscible with water such as acetone, dioxane, and dimethyl sulfoxide, and mixtures thereof.

The raw material (n) is easily derived from glutaric acids (II/)''.

(VI) (n) [wherein, X is halogen (e.g., chlorine, bromine, etc.), Z
represents an amine protecting group (e.g., carbobenzoxy, t-butoxycarbonyl, and AII, AII, and R have the same meanings as above.) Synthesis of 7-hydroxyglutamic acid (i) T
, Kaneko et al., Bull. Chem. Soc.
．． Jap. , 35.

Synthesis of 7-alkylglutamic acids (i) For example, J. Done et al., Biocham.
J. , 51. 451 (1952), according to the method for synthesizing 7-methylglutamic acid.

Synthesis of 7-aminoglutamic acids (i) Chemical abstract Zoo (18):
145862a Manufacturing method B (In the formula, AI'', A'', R1 and Hal have the same meanings as above, and As represents an ester residue.) In the above manufacturing method B, the first step and the second step are each a manufacturing method. What is necessary is just to carry out similarly to the 1st process and 2nd process in A.

Third step is a step in which compound (IX) is decarboxylated to obtain target compound (Ib).

Decarboxylation may be carried out according to conventional methods such as heating at about 40 to about 100'C in an aqueous solvent, and in order to accelerate the reaction,
A suitable acid (eg, hydrochloric acid, hydrobromic acid, etc.) can also be added.

The raw material (■) is a compound (X) and an acrylic acid derivative (X
I) Michael addition reaction with th
It can be easily obtained by following the steps below.

Tei 2) 'Modern Synthetic
Reactions JH. O. House.

2nd ed. ;W. A. Benjamin:
Menlo park. C.A.

p9595-623 (1972) (Hereinafter in the margin) (In the formula, A'', A1, A1, R, and Z have the same meanings as above.) Target substance (I) has two asymmetric carbon atoms and has a diastereo Although it is produced as mer and optical isomer, these can be easily optically resolved by conventional methods, and both are subject to the present invention.

An antitumor test of target substance (I) against L1210 was carried out in the same manner as described in JP-A-57-225272, and N-[4-[[
(2,4-diamino-6-buteridinyl)methyl]methylamino]benzoylco-7-methylglutamic acid U is rLss. -10mg/kg; IL 5uAx-2
00mg/kg; showed CI-20.

In addition, target substance (I) exhibits lower toxicity than methotrexate and also has excellent anticancer effects, so it can be said to be useful as an anticancer agent.

In addition, the target substance (I) can also be used for rheumatoid arthritis, erythematosus erythematosus, etc., and autoimmune diseases.

The object substance (I) of the present invention can be administered orally or parenterally to humans or animals. For example, compound (I) can be administered orally in the form of tablets, granules, powders, capsules, liquids, etc., or parenterally as injections, etc. These preparations can be manufactured according to well-known methods using additives such as excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents.

When using target substance (I) for tumor treatment in adults-11
~500 mg may be administered orally or parenterally once a day or in several divided doses, but it is preferable to administer an amount appropriate to the patient's gender, age, medical condition, etc.

Embodiments of the present invention are shown below.

Example 1 (a) To a solution of 276 mg of 1 (isomer A:B-2, 3:1) dissolved in 2.8 ml of dimethylacetamide, 2.4
- Add 308 mg of diamino-6-bromomethylpteridine hydrobromide/isopropanol adduct 11",
Stirred at 0-55°C for 8 hours. Chloroform 120ml
Then, 120 ml of 0.3N aqueous ammonia was added and stirred for 10 minutes, and the chloroform layer was separated. It was washed with water for 3 days, dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue was chromatographed on silica gel using a Lobar column and eluted with chloroform:methanol (20: lv/v). Concentrate the eluate and
Dimethyl N-[4-[[(2,4-diamino-6-buteridinyl)methyl]methylamino]benzoylco-7-
2354 mg of methylthioglutamate was obtained. Yield, 8
4.5%, melting point: 134-138"C (recrystallized from acetonitrile-methanol).

IR(CHCL3> 3400.2980.1730.
1650.1605゜1490, 1440.1300-
1160 am-'NMR (CDC1s"CD5OD)
82.14 (s, 3H).

2.20-2.47 (m, 2H), 3.1B
(s, 3H), 3.334.74 (s, 2H)
, 4.93 (m IH), 6.75 (d,
2H.

J=9.0Hz>, 7.70 (d, 2H, J=9
．． o) (z), 8.59 (s, IH) (b) Add 2.4 ml of IN sodium hydroxide aqueous solution to a solution in which 2355 mg of the above was suspended in 12 ml of ethanol,
The mixture was stirred at room temperature for 24 hours. After cooling with water, the precipitate was collected by filtration, washed with ethanol, and dissolved in 5 ml of water, and the pH of the solution was adjusted to pH 4 with IN hydrochloric acid. After leaving it in the refrigerator overnight,
The deposited precipitate was collected by filtration, and N-[:4[[(2,4-diamino-6-buteridinyl)methyl]methylamino]benzoyl core methylthioglutamate (isomer A:B
=3.7:1) 290 mg was obtained. Yield: 81.9%,
Melting point: 192-199°C (decomposed).

IR(KBr) 3600-3100.1640.16
00.1505°1440.1365.1245.12
05 am””NMR (DMSO-ds) 81.77
-2.30 (m, 2H), 2.05 (s, 3H),
3.20 (m, IH), 3.21 (s, 3H)
, 4.63 (m, 18), 4.80 (s, 2
H), 6.80 (s, 2H), 6.84/
J QLI v-n /1u-S ri 7ri/nii,, riu1 ri A (d, 2H, J=
9.0Hz), 8.21 (d, IHJ=7.5
Hz).

8.60 (s, 1) 1) Elemental analysis (as C,, H*, 0JaS・2HgO) Calculated value (%): C, 47,01; H, 5,26;
N, 20.88; S, 5.98 Experimental value (%'): C, 47,41; H, 5,21
i N, 21.10; S, 5.97 ml) 2,4-Diamino-6-bromomethylpteridine was synthesized by J, Org, Chem, 42.
208.1977.

In addition, in the examples and reference side, isomer A or isomer B
represents 20 Eri bodies or Threo bodies.

Example 2 (a) To a solution of 93 mg of 4 (isomer B) dissolved in 1 ml of dimethylformamide, 146 mg of 2,4-diamino-6-promomethylbuteridine hydrobromide/impropatur adduct was added, Stirred at 50-55°C for 24 hours. The reaction solution was treated in the same manner as in Example 1-(a) to give diimpropyl 7-amino-N-[4-[:[:(2,4-diamino-6-buteridinyl)methylcomethylamino]benzoylcoglutamate. 585 mg was obtained. Yield: 61.
6%, melting point: 121-123°C (recrystallized from acetonitrile-methanol).

IR(CHCIm) 3510.3400.2975.
1725.1640°1605, 1495.1440.
1375°1290-1160.1105 cm-'N
MR (CDC1m+CD5OD) 81.10-1.3
7 (m, 12H).

1.90-2.57 (m, 2H), 3.16 (s
, 3H), 3.53 (dd, LH, J=8.1Hz,
, C4,5Hz>, 4.72 (s, 2H).

4.63-5.22 (m, 3H), 6.73 (
d, 2H,, c9.0Hz).

7.71 (d, 2H, J=9.0Hz), 8.5
8 (s, IH) elemental analysis (C*5HsaO, No.0
．． 5H, O) Calculated value (%): C, 55, 50
: H, 6, 45: N, 22.41 Experimental value (%):
C, 55,28; H, 6,48S N, 22.29
6237 mg of 11M main (a) and 2.4 ml of dimethylacetamide
195 mg of 2,4-diamino-6-brol adduct was added to the solution dissolved in , and stirred at 50 to 55°C for 9 hours.

The reaction solution was treated in the same manner as in Example 1(a), and diethyl N
-[4-[:[(2,4-diamino-6-pteridinyl)methylcomethylaminocobenzoyl]-7-benzoyloxy-α-ethoxycarbonylglutamate 726
6 mg was obtained. Yield: 82.4%, melting point = 125-12
7°C (chloroform-ether color recrystallization), 'IR (CIC1,) 3500.3400.2970.
1730.1650°1600, 14g0.1435.
1370.1305゜1280-1170.1100
am'″lNMR (CDC1m+CD5OD) & 1.
00-1.35 (m, 9) 1).

2.97-3.43 (m, 2H), 3.19 (s
, 3H), 3.87-4.40 (m, 6H), 4.
75 (s, 2H), 5.41 (dd, IH.

J=8.1Hz, J=4.5Hz), 6.74 (d
, 2H, J=9.0Hz).

7.23-7.63 (m, 3H), 7.69 (d
, 2H, J=9.0Hz>.

7.97 (dd, 2H, J=7.5Hz, J=1.5
Hz), 8.61 (s, IH) (b) 1637 mg of the above product was dissolved in 50 ml of ethanol, and the mixture was stirred at room temperature for 28 hours. After cooling with water, the precipitate was collected by filtration, washed with ethanol, and dissolved in 10 ml of water, and the solution was adjusted to pH 4 with IN hydrochloric acid. After standing in the refrigerator overnight, the precipitate that precipitated was collected by filtration. 22 ml of water was added and the suspended solution was adjusted to pH 1 with IN hydrochloric acid. 50-55℃
After heating and stirring for 4 hours and cooling, the pH was adjusted to 10 with IN aqueous sodium hydroxide solution, filtered, and the filtrate was adjusted to pH 4 with IN hydrochloric acid. After cooling with water, the precipitate that precipitates out is collected by filtration, and N-[a-[[
943 mg of (2,4-diamino-6-buteridinyl)methyl]methelaminocobenzoyl-7-hydroxyglutamic acid (isomer A:B-1,4:1) was obtained. Yield near 7.9%, melting point: 240-250°C (decomposed).

IR(KBr) 3600-3100.1630.16
00.1505°1360, 1240.1200.11
05 am-'NMR (DMSO-do) δ1.73-
2.27 (m, 2H), 3.20 (s, 3H),
3.80-5.20 (m, 2H), 4.79 (s
, 2H).

6.83 (d, 2H, J=9.0Hz), 6.87
(s, 211), ? ,60(brs,2)1),
7.73 (d, 2H, J=9.0Hz), 8.28
(d, LH, J=7.51(z), 8.61 (
s, LH) Elemental analysis (C!.Hz *OaNm・3.5
Calculated value (%): C, 45,03;
H, 5, 48; N, 21. OO experimental value (%): C
, 45,09; H, 5,08; N, 20.78 Example 4 (a) 93343 mg in 33 ml of dimethylacetamide
2,4-diamino-6-promomethylbuteridine hydrobromide-impropatool adduct 34 in a solution dissolved in
46 mg was added and stirred at 50-55°C for 8 hours. The reaction solution was treated in the same manner as in Example 1(a), and diethyl N-[-
103,869 mg of [:[(2,a-diamino-6-buteridinyl)methylcomethylaminocobenzoyl]-α-ethoxycarbonyl-7-methylglutamate was obtained.

Yield near 9.6%, melting point: 178-179.5°C (recrystallized from chloroform-ether).

IR(CHCl,) 3510.3400.2975.
1730.1650°1600, 1485.1440.
1375.1320-1160, 1090.1000.
825 am-'NMR (CDC1, +CD5OD)8
0.97-1.37 (m, 12H).

2.23-3.07 (m), 3.21 (s, 3H
), 3.73-4.40 (m, 6H), 4.77
(s, 2H), 6.78 (d, 2H.

J=9.0Hz>, 7.68 (d, 2H, J=9.
0 Hz), 8.60 (s, LH) (b) Add 13 ml of IN sodium hydroxide aqueous solution to a solution of 101,628 mg of the above suspended in 60 ml of ethanol. - After standing overnight, the deposited precipitate was collected by filtration. The solution in which 16 ml of water was added and suspended was adjusted to pH 1 with IN hydrochloric acid, heated and stirred at 80° C. for 3.5 hours, and after cooling, the pH was adjusted to 4 with IN aqueous sodium hydroxide solution. After cooling with water, the precipitate precipitated was collected by filtration, and N-[-
[:[:(2,4-diamino-6-buteridinyl)methylcomethylaminocobenzoyl-7-methylglutamine 11 (isomer A:B=1:1.4) 1152 mg was obtained.

Yield: 84.6%, melting point 2189-194°C (decomposed).

IR(KBr) 3600-3100.1640.16
00.1510゜1450,,1360.1305.1
200.830°765 am-' NMR (DMSO-ds) 81.10 (d, 3H, J
=6.6Hz>.

1.57-2.63 (m, 3H), 3.21 (s
, 3) 1), 4.37 (m, IH), 4.80 (
s, 2H), 6.81 (d, 2H.

J=9.0Hz), 7.09 (s, 2H), 7.
72 (d, 2H.

J=9.0)lz), 8.00 (brs, 21()
, 8.18 (d, IH.

J=7.5Hz), 8.61 (s, IH) elemental analysis (C! IH! 40aks・2.5H*O) Calculated value (%): C,49,12; )1,5.69
F N, 21.82 Experimental value (%): C, 48,95
; H, 5,65; N, 21.52 Reference Example 1 (Cbz: Carbobenzoxy) (a) 4.3 ml of thionyl chloride was added to a mixture of 12,824 mg of 7-methylthioglutamic acid and 16 ml of methanol, and the mixture was heated at 40"C for 4. The mixture was heated and stirred for 5 hours. After cooling, the solvent was distilled off from the reaction mixture, methanol was added, and the mixture was concentrated to obtain 131,090 mg of dimethyl 7-methylthioglutamate hydrochloride as an oil.

NMR (CDC1*”CD5OD) & 2.13-2.
80 (m, 2H).

2.19 (s, 3H), 3.33 (m, I
H),, 3.77 (s, 3H).

3.86 (s, 3H), 4.30, (m, LH>
(b) 131090 mg of the above and 14 dimethoxyethane
1.46 g of N-carbobenzoxy-N-methyl-4-aminobenzoyl chloride was added to the suspension consisting of ml and 1.37 ml of triethylamine was added dropwise under water cooling, followed by stirring for 4 hours.

Ethyl acetate is added to the reaction mixture, which is washed successively with dilute hydrochloric acid, water, dilute aqueous sodium bicarbonate solution, and water, dried over anhydrous sodium sulfate, and concentrated. The residue was subjected to silica gel chromatography using a Roper column and benzene:ethyl acetate (
-3: eluted at 1v/v). The eluate was concentrated and dimethyl N-C4-CCcarbobenzoxy)methylaminocobenzoyl-7-methylthiazglutamyl) 14133
Obtained 0 mg as an oil. Yield: 63.9
%.

IR(CHClj) 2950.1730.1700.
1660.1605°1490.1430.1380.
1345.1310-1180.1150.1110,
1100°850 cm-' NMR (CDCIs) & 2.00-2.73 (m
, 2H), 2.15 (s, 3H), 3.33 (s
, 3H), 3.35 (m, 1)l).

3.62.3.72. and 3.77 (3s(
erythro.

three), 6H), 4.97 (m, IH),
5.16 (s, 2H).

6, 78 and 6.92 (2d(erythro
, three), LH.

J=7.5Hz), 7.31 (s, 5H),
7.34 (d, 2H.

J=9.0Hz), 7.77 (d, 2H, J=9
．． 0Hz) (C) Add 30% hydrogen bromide to 1.30g of the above mouth.
7.5 ml of acetic acid was added, and the mixture was stirred at room temperature for 4 hours. 75 ml of ether was added to the reaction solution, the ether solution was removed by decanting while cold, and the remaining precipitate was washed with ether and dissolved in methylene chloride. The methylene chloride solution was sequentially washed with an aqueous potassium hydrogen INi acid solution and water, dried over anhydrous sodium sulfate,
Concentrated. The residue was subjected to silica gel chromatography using a Roper column using benzene:ethyl acetate (2:
lv/v). The eluate was concentrated and dimethyl N-
[4-do shi 4-1 shi〒 q ) ) Beno 1/no
11-] -9-314-+1-4-naglutamate (1746 mg) was obtained as an oil.

The yield was near 9.1%.
il) The sample was subjected to HPLC using an ODS column and eluted with methanol:water (1:l'v/v). Concentrate the eluate to obtain isomer A:B-2, 3:1 components and isomer A:B.
-1:2.3 components were obtained.

IR(CHClj) 3440.2960.1730.
1650.1610°1495, 1440.1335.
1310-1170°835 cm-' NMR of Isomer A (CDCIs) & 2.13 (s,
3H), 2.20-2.47. (m, 2H), 2.8
3 (s, 3H), 3.33 (t, LH.

J-6,8Hz), 3.60 (s, 3H), 3.
73 (s, 3H).

4.97 (m, LH), 6.54 (d, 2)1.
J=9.0Hz), 6.60(d, IH, J=7.5
Hz>, 7.64 (d, 2H, J=9.0Hz) Isomer B (') NMR (CDCIs >81.98-2
．． 78 (m, 2H).

2.13 (s, 3H), 2.84 (s, 3H),
3.33 (dd, 1.H.

J=8.4! I=,, J=6.0Hz), 3.70
(s, 3H), 3.75 (s, 311), 4.87
(m, IH), 6.55 (d, 2H.

J=9.0Hz), 6.69 (d, LH, J=7.
5Hz), 7.64(d, 21., C9, OHz
) Part 2 Main (a) 7-Amine Glutamic Acid/0.5 Hydration Story 710
4.2 ml of thionyl chloride was added to a mixture of 15 ml of Impropatol and the mixture was heated under reflux for 20 hours. The reaction solution was treated in the same manner as in Reference Example 1(a) to obtain 1.32 g of diisopropyl 7-aminoglutamate dihydrochloride as a foamy substance.

NMR (CDCIS+CD5OD>Sl, 35 (d,
12H, J=6.0Hz).

2.70 (m, 2H), 4.53 (m, 2H),
5.16 (m, 2H) (b) A suspension consisting of 16,332 mg of the above and 4 ml of dimethoxyethane was heated to -30°C.
316 mg of N-carbobenzoxy-N-methyl-4-aminobenzoyl chloride was added thereto, followed by 0.51 ml of triethylamine, and the mixture was stirred for 30 minutes. The reaction solution was treated in the same manner as in Reference Example 1(b), and 137 mg of isomer A and 137 mg of isomer B of diisopropyl 7-amino-N-[4-[(carbobenzoxy)methylaminocobenzoyl]glutamate 17 were added. Obtained as an oil. Yield from long grain, 51.4%.

Isomer A: IR (CHCIm) 3400.2970.1?25.
1700.1655°1605, 1495.1375.
1350.1290-1140, 1105 cm-' NMR (CDCIs) δ1.06-1.40 (m, 1
2H), 1.70-2.50 (m, 2H), 1.
81 (s, 2H), 3.34 (s, 3) 1).

3.53 (dd, IH, J=;12.0HzJ=3.
0Hz), 4.77-5°, 30 (m, 3H),
5.17 (s, 28), 7.33 (s, 5H).

7.34 (d, 2HJ=9.0Hz), 7.
84 (d, 2H.

J=9.0Hz), 8.61 (d, IH, J=7.
5Hz) Isomer B: IR (CHCl, > 3400.2975.1725.
1700.1660°1605, 1495.1375.
1345.1290-Li2O, 1105 am-' NMR (CDCIn) 81.06-1.40 (m,
12H), 1.80-2.60 (m, 2) 1), 1.
81 (s, 2H), 3.33 (s, 3H).

3.52 (dd, LH, J-9,0Hz, J=4.5
Hz), 4.37-5.27 (m, 3H), 5.
17 (s, 2H), 7.05 (d, IH.

J=7.5Hz), 7.33 (s, 5H), 7.
34 (d, 2H.

J=9.0Hz), 7.84 (d, 2H, J: 9.
01 (z) (c) 30 to 169 mg of the above ■ (isomer B)
1.5 ml of % hydrogen bromide-acetic acid was added, and the mixture was stirred at room temperature for 4 hours. The reaction solution was treated in the same manner as in Reference Example 1(C) to obtain 498 mg of diisopropyl 7-amino-N-[4-(methylamino)penzoylcoglutamate as a foamy substance. Yield near 8.4%.

IR(CHCIm) 3450.2980.1735.
1650.1610°1505, 138G, 1335
．． 1300-1170°1100+ 840 cm-' MILD /rM1jr 1 10-I QF
(m 1')I) 1 an-') 6n(m
, 2H), 2.85 (s, 3H), 2.93 (
s, 2M).

3.53 (m, 11), 4.65-5.30 (
m, 3H), 6.55 (d, 2H, J=9.0Hz
>, 7.36 (d, IH, J=7.5Hz).

7.68 (d, 2H, J=9.0Hz>(a) 艮3.
A solution of 16 mg of sodium dissolved in 1.3 ml of ethanol was added to a suspension of 0.07 g in 1 ml of ethanol, and then a solution of 191.59 g dissolved in 0.5 ml of ethanol was added, followed by stirring at room temperature for 40 minutes. Ethyl acetate was added to the reaction solution, which was washed successively with dilute hydrochloric acid, water, dilute aqueous sodium bicarbonate solution, and water, dried over anhydrous sodium sulfate, and concentrated.

The residue was subjected to silica gel chromatography using a Roper column and eluted with benzene:ethyl acetate (6:IV/V). The eluate was concentrated to obtain 204.15 g of diethyl 7-benzoyloxy-N-[4-[(carbobenzoxy)methylamine]benzoyl]-α-ethoxycarbonylglutamate as an oily substance. Yield, 90.4
%.

IR(CHCl,) 3400.2970.1735.
1700.1665°1600, 1480.1345.
1315.1290-1180, 1155.1105.
1015°850 cm-' NMR (CDCIs) & 1.03-1.37 (, m,
9H), 3.02-3.55(m, 2H), 3
．． 34 (s, 3) 1), 3.90-4.43
(m, 6H).

5.19 (s, 2H), 5.37 (dd, IH,
J=8.4Hz.

J=4.2Hz>, 7.17-7.60 (m, 4H
), 7.34 (d.

2B, J: 9.0Hz), 7.36 (s, 5H),
7.80 (d, 2H.

J=9.0Hz), 7.97 (dd, 2H, J=7
．． 5Hz, J=1.5)1z) (b) 387mg of the above
3.3 ml of 30% hydrogen bromide-acetic acid was added to the mixture, and the mixture was stirred at room temperature for 3 hours. The reaction solution was treated in the same manner as in Reference Example 1(c) to obtain 6269 mg of diethyl 7-benzoyloxy-α-ethoxycarbonyl-N-[4-(methylamino)penzoylcoglutame h as an oily substance. Yield: 8
7.1%.

IR (CHCIs) 3410.2980.1?35.
1655.1610°1480.1370. 1310
．． 1280-1170°1105, 1020.83
0 cm-'NMR (CDC1,) Sl, 00-1.3
5 (m, 9H), 2.73 (s, 3H), 2.9
8-3.48 (m, 2H), 3.87-4.37 (
m, 6H), 5.41 (dd, IH, J=8.1H
z, J=4.5Hz).

6.52 (d, 2H, J=9.0Hz), 7.15
-7.60 (m, 4H).

7.64 (d, 2H, J=9.0Hz>, 7.97
(dd, IH.

J = 7.5 Hz, JJ, 5 Hz > (Left below) Vines 2 Kuto (a) Add 184.43 g to a solution of 23 mg of sodium dissolved in 4 ml of ethanol, and add 1.86 m of gain.
1 and stirred at room temperature for 1 hour.
Treated in the same manner as in (a) to obtain diethyl N-44-[(carbobenzoxy)methylamino]hensoyl-α-ethoxycarbonyl-7-methylglutamate 6, yield: 96.4%.

IR (CHCIS)3400.2960,1725.
1695.1655.

1600, 1485. 1375, 1340.
1320-1130, 1090.1000.850
cm-'NMR (CDCIs) Sl. 02-1.37
(m.12H), 2.20-3.10 (m.3H),
3.35 (s.3H), 3.78-4.43
(m.6H).

5, 19 (s, 2H), 7.36 (s. 5H)
, 7.38 (d.2H.

J=9.0Hz>, 7.49 (s.IH),
7, 82 (d.2H.

J=9. 0Hz) (b) 5.37g of the above name and 27m of 30% hydrogen bromide-acetic acid
1 was added thereto, and the mixture was stirred at room temperature for 4 hours. Reference example 1
Treated in the same manner as (c), α-ethoxycarbonyl-7-
113.45 g of methyl-N-[4-(methylamino)benzoyl]glutamate was obtained. Yield: 84.7%, melting point = 65-66°C (recrystallized from ether-hexane).

IR (CHCII) 3410. 2970, 17
30, 1650. 1605.

1485, 1320-1150. 1090. 83
0 am”NMR (CDC1s”CDsOD) δ 1
．． 03-1.37 (m.12H).

Claims (1)

[Claims] General formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, A^1 and A^2 are each hydrogen or ester residue; R is C_1 to C_5 alkyl, C_1 to C_
5 represents alkylthio or hydroxy. ) or its salt.