JPH06107546A - Adm resistance-dissolving medicine - Google Patents

Abstract

PURPOSE:To provide an adriamycin resistance-dissolving medicine containing a specified compound having a base skeleton composed of tyrosine as the active component, capable of overcoming adriamycin resistance and amplifying the effect of adriamycin. CONSTITUTION:An adriamycin(ADM) resistance-dissolving medicine containing a compound of formula I [R1 is 5-isoquinolyl or p-aminoethyloxyphenyl; R2 and R3 are each H or methyl; (n) is 2 or 3: R4 is benzyloxycarbonyl or 3- phenylpropionyl] or its pharmaceutically permissible acid adduct as the active component. The compound of formula I, is exemplified by e.g. N-[2-(4- benzyloxycarbonylpiperazino)-1-(p-methoxybenzyl)ethyl]-N-methyl-p-2- aminoethyloxybenzenesulonic acid amide. The compound of formula I, e.g. a compound of formula II can be obtained by reacting tyrosine of formula III with piperazine and then carrying out condensation reaction between the resultant compound of formula IV and isoquinolinesulfonic acid chloride.

Description

Detailed Description of the Invention

[0001]

The present invention relates to adriamycin (AD
M) It relates to an ADM resistance-eliminating agent containing a derivative having tyrosine as a basic skeleton which acts on a cancer cell in a resistant strain and enhances the effectiveness of ADM treatment.

[0002]

2. Description of the Related Art Conventionally, drugs for the purpose of releasing resistance of ADM-resistant strains have been synthesized and clinically used, but still satisfactory therapeutic effects have not been obtained.

[0003]

DISCLOSURE OF THE INVENTION The object of the present invention is to provide a derivative which overcomes ADM resistance and enhances the action of ADM.

[0004]

Means for Solving the Problems The present invention will be described in brief. The present invention relates to an ADM resistance releasing agent, which is represented by the following general formula (Formula 1):

[0005]

[Chemical 1]

(Wherein R ₁ is a 5-isoquinolyl group or p
-Aminoethyloxyphenyl group, R ₂ and R ₃ represent hydrogen or methyl group, n represents 2 or 3, and R ₄ represents benzyloxycarbonyl group or 3-phenylpropionyl group) or a compound thereof. It is characterized in that a physiologically acceptable acid addition salt is used as an active ingredient.

The inventors of the present invention have been keenly searching for a compound that overcomes ADM resistance and enhances the action of ADM. As a result, the present inventors have previously found that vascular smooth muscle relaxation and platelet aggregation inhibition which act mainly on the circulatory system. A compound having an effect was developed, and the Akemitsu was found in a part of the derivative having tyrosine as a basic skeleton filed as Japanese Patent Application No. 2-52686, and the present invention was completed.

Specific compounds represented by the general formula (Formula 1) of the present invention include the following compounds, but the present invention is not limited to these compounds. (1) N- [2- (4-benzyloxycarbonylpiperazino) -1- (p-methoxybenzyl) ethyl] -N
-Methyl-p-2-aminoethyloxybenzenesulfonic acid amide (2) N- [2- (3-phenylpropionylhomopiperazino) -1- (p-hydroxybenzyl) ethyl]-
5-Isoquinolinesulfonic acid amide (3) N- [2- (4-benzyloxycarbonylpiperazino) -1- (p-methoxybenzyl) ethyl] -N
-Methyl-5-isoquinoline sulfonic acid amide

[0009]

[Table 1]

Examples of the acid addition salt of the isoquinolinesulfonic acid derivative represented by the general formula (Formula 1) include inorganic acids such as phosphoric acid, hydrochloric acid and sulfuric acid, or acetic acid, citric acid, succinic acid, fumaric acid, tartaric acid and the like. The salts of organic acids can be mentioned.

The compound represented by the general formula (Formula 1) of the present invention can be synthesized by the method described in Japanese Patent Application No. 2-52686. The method for producing the compound of the present invention described in the specification is roughly the following method.

A known compound, a tyrosine of the following general formula (Formula 2) protected with an amino group:

[0013]

[Chemical 2]

One of the following general formulas (Chemical Formula 3) obtained by reacting one nitrogen-protected piperazine:

[0015]

[Chemical 3]

The free amino compound obtained by removing the amino-protecting group (1) is condensed with isoquinolinesulfonic acid chloride to give the following general formula (Formula 4):

[0017]

[Chemical 4]

Was obtained. Further, if desired, in this compound (Chemical Formula 4), (a) a step of liberating a hydroxyl group by hydrolysis, (b) a step of eliminating the protecting group of piperazine,
(C) methylating a free hydroxyl group, (d) acylating a piperazine, and (e) methylating a sulfonic acid amide. In addition, the carbonyl group is reduced to a methylene chain before or after any of the steps of the above general formula (Formula 4) or (a) to (e). 1 or 2 of these operations
By carrying out the above, some of the compounds of the present invention can be produced.

Homopiperazine can be used in place of the piperazine used in this example. In the general formula (Formula 2), the protected amino group may be substituted with methyl, or the hydroxyl group may be protected.

Next, a production example of the above-mentioned compound (3) will be described. Production Example 1 [Compound (3)] O-benzyl-N-benzyloxycarbonyltyrosinol O-benzyl-N-benzyloxycarbonyltyrosine methyl ester 27.25 g was added to ethanol 185 ml,
The mixture was dissolved in 122 ml of a mixed solvent of tetrahydrofuran, 5.8 g of lithium chloride and 5.2 g of sodium borohydride were added under ice cooling, and the mixture was stirred at room temperature for 18 hours. After adding 500 ml of saturated saline to the reaction solution, chloroform 3 was added.
The product was extracted twice with 00 ml, dried over magnesium sulfate, and concentrated under reduced pressure to obtain 146 mg of the desired product as colorless crystals. IR (KBr) cm ⁻¹ : 1620, 1510, 144
0, 1320, 1160

1- [2-benzyloxycarbonylamino-3- (p-benzyloxyphenyl) propyl]-
Crystals of 4- (t-butoxycarbonyl) piperazine O-benzyl-N-benzyloxycarbonyltyrosinol (6.0 g) were dissolved in carbon tetrachloride (70 ml), triphenylphosphine (4.5 g) was added, and the mixture was heated under reflux for 20 hr. The insoluble material was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was applied to a silica gel column, and hexane-ethyl acetate (6:
Purification in 1) yielded 5.02 g of colorless crystals of 2-benzyloxycarbonylamino-3- (p-benzyloxyphenyl) propyl chloride. 4.5 g of this crystal is added to N
-(T-Butoxycarbonyl) piperazine was dissolved in 40 ml of dimethylformamide, 1.8 g of methyl iodide and 1.66 g of potassium carbonate were added, and the mixture was stirred at 120 ° C for 3 hours. 100 ml of saturated saline solution was added thereto, extracted twice with 60 ml of chloroform, dried over magnesium sulfate and concentrated under reduced pressure. The resulting residue was purified with hexane-ethyl acetate (2: 1) to obtain the colorless amorphous target product 2. 74 g was obtained. ¹ H-NMR (CDCl ₃ , δppm): 1.45 (9
H, s), 2.82 (2H, m), 3.36 (4H,
m), 5.09 (2H, s), 7.06 (2H, d, J
= 8.3 Hz), 7.3 to 7.45 (5H, m)

N- {2- [4- (t-butoxycarbonyl) piperazino] -1- [p- (5-isoquinolylsulfonyloxy) benzyl] ethyl} -5-isoquinoline sulfonic acid amide 1- [2- Benzyloxycarbonylamino-3- (p
-Benzyloxyphenyl) propyl] -4- (t-butoxycarbonyl) piperazine (1.7 g) in methanol 2
Dissolve in 5 ml, add 5% palladium / carbon (1.0 g), stir in a hydrogen atmosphere for 20 hours, separate insoluble matter by filtration, concentrate the filtrate under reduced pressure, and remove the residue in tetrahydrofuran (30).
ml was added and dissolved, 2.8 g of 5-isoquinolyl sulfonyl chloride.hydrochloride and 4 ml of triethylamine were added thereto under ice cooling, and the mixture was stirred at room temperature for 3 hours. After adding 100 ml of water to the reaction mixture, the mixture was extracted twice with 70 ml of chloroform, dried over magnesium sulfate and concentrated. The residue was applied to a silica gel column and chloroform-methanol (100: 1).
~ 50: 1) to obtain 1.24 g of a yellow amorphous target product. ¹ H-NMR (CDCl ₃ , δppm): 1.40 (9
H, s), 1.75 to 2.18 (6H, m), 2.15
(6H, m), 3.27 (1H, m), 5.35 (1
H, br), 6.67 (2H, d, J = 8.3Hz),
7.65 (1 H, t, J = 7.8 Hz), 7.69 (1
H, t, J = 7.8 Hz), 8.21 (1H, d, J =
8.3 Hz), 8.27-8.32 (2H, m)

N- {2- [4- (t-butoxycarbonyl) piperazino] -1- [p- (5-isoquinolylsulfonyloxy) benzyl] ethyl} -N-methyl-5-
Isoquinoline sulfonic acid amide N- {2- [4- (t-butoxycarbonyl) piperazino] -1- [p- (5-isoquinolylsulfonyloxy) benzyl] ethyl} -5-isoquinoline sulfonic acid amide About 1000 mg of tetrahydrofuran And dimethylformamide (7.5 ml each) were dissolved, and 60% sodium hydride (67 mg) and methyl iodide (0.1 mg) were added under ice cooling.
11 ml was added, and the mixture was stirred at room temperature for 1 hour. 30 ml of saturated saline was added to the reaction solution and extracted with 40 ml of ethyl acetate. The extract was washed with saturated saline, dried over magnesium sulfate and concentrated under reduced pressure. The residue was applied to a silica gel column and chloroform-methanol (60: 1). ) To obtain 742 g of a yellow amorphous target product. ¹ H-NMR (CDCl ₃ , δppm): 1.44 (9
H, s), 2.21 (5H, m), 2.40 (1H, d
d, J = 6.9, 12.6 Hz), 3.17 (9H,
m), 4.12 (1H, m), 6.60 (2H, d, J
= 8.8 Hz), 7.58 (1H, t, J = 7.8H
z), 7.63 (1H, t, J = 7.8Hz), 8.1
3 (1H, d, J = 8.3 Hz), 8.21 to 8.30
(4H, m)

N- {1- [p- (5-isoquinolylsulfonyloxy) benzyl] -2- [4-benzyloxycarbonylpiperazino] ethyl} -N-methyl-5-isoquinolinesulfonic acid amide N- {2- [4- (t-butoxycarbonyl) piperazino] -1- [p- (5-isoquinolylsulfonyloxy) benzyl] ethyl} -N-methyl-5-isoquinolinesulfonic acid amide 4N chloride in about 742 mg 10 ml of an ethyl acetate solution of hydrogen was added, the mixture was stirred at room temperature for 1 hour and concentrated under reduced pressure, 30 ml of a saturated aqueous solution of sodium bicarbonate was added, and the mixture was extracted twice with 20 ml of a mixed solvent of chloroform-isopropyl alcohol (5: 1). The extract was dried over magnesium sulfate and concentrated under reduced pressure to give 10 m of methylene chloride.
It was dissolved in 1 l, benzylchloroformate 0.29 ml and triethylamine 0.4 ml were added under ice cooling, and the mixture was stirred for 2 hours under ice cooling, saturated saline 40 ml was added, and the mixture was extracted twice with 20 ml of chloroform for dry extraction. Was dried over magnesium sulfate and concentrated under reduced pressure, and the residue was applied to a silica gel column, chloroform-methanol (60: 1).
Purification was performed to obtain 665 mg of a pale yellow amorphous product. ¹ H-NMR (CDCl ₃ , δppm): 2.18-
2.28 (5H, m), 2.37 (1H, dd, J =
7.3, 12.7 Hz), 2.83 (3H, s), 6.
61 (2H, d, J = 8.8Hz), 7.59 (1H,
t, J = 7.8 Hz), 7.63 (1H, t, J = 7.
8Hz), 8.12 (1H, t, J = 8.3Hz),
8.23-8.35 (4H, m)

N- [2- (4-benzyloxycarbonylpiperazino) -1- (p-hydroxybenzyl) ethyl] -N-methyl-5-isoquinolinesulfonic acid amide N- {1- [p- (5 -Isoquinolylsulfonyloxy) benzyl] -2- [4-benzyloxycarbonylpiperazino] ethyl} -N-methyl-5-isoquinolinesulfonic acid amide (about 650 mg) was dissolved in 6 ml of methanol, and 30 ml of 1N caustic soda solution was added. Add mixed solvent of chloroform-isopropyl alcohol (5: 1) 2
Extracted twice with 0 ml. The extract was dried over magnesium sulfate, and the residue was concentrated under reduced pressure and subjected to silica gel column, chloroform - methanol (80: 1 to 50: 1) ¹ to give the desired product 329mg of yellow MuAkirajo purified by H-NMR (CDCl ₃ , δppm): 2.39 ~
2.55 (6H, m), 2.64 (1H, dd, J =
6.4, 12.7 Hz), 2.87 (1H, dd, J =
4.4, 14.6 Hz), 2.97 (3H, s), 3.
44 (4H, m), 4.03 (1H, m), 5.13
(2H, s), 6.28 (2H, d, J = 8.3H
z), 6.62 (2H, d, J = 8.3 Hz), 7.3
6 (5H, s), 7.39 (1H, dd, J = 7.3,
7.8 Hz), 8.09 (1H, d, J = 5.9H
z), 8.13 (1H, d, J = 7.8Hz), 8.2
6 (1H, d, J = 7.3 Hz), 8.48 (1H,
d, J = 5.9 Hz), 9.27 (1H, s)

N- [2- (4-benzyloxycarbonylpiperazino) -1- (p-methoxybenzyl) ethyl] -N-methyl-5-isoquinolinesulfonic acid amide N- [2- (4-benzyloxy) Carbonylpiperazino) -1- (p-hydroxybenzyl) ethyl] -N-
Methyl-5-isoquinoline sulfonic acid amide About 200m
g was dissolved in 5 ml of chloroform, 1 ml of methanol was added, an ether solution of diazomethane was added under ice cooling, and the mixture was stirred for 1 hour and 30 minutes. The reaction mixture was concentrated, the residue was applied to a silica gel column, and chloroform-methanol (80:
1 to 50: 1) and purified to be yellow amorphous substance 101 mg
Obtained ¹ H-NMR (CDCl ₃ , δppm): 2.32-
2.45 (5H, m), 2.53 to 2.65 (2H,
m), 2.78 to 2.90 (1H, m), 2.92 (3
H, s), 3.37 (4H, m), 3.73 (3H,
s), 4.16 (1H, m), 5.11 (2H, s),
6.50 (2H, d, J = 8.3Hz), 6.82 (2
H, d, J = 8.3 Hz), 7.35 (5 H, s),
7.55 (1 H, t, J = 7.8 Hz), 8.09 (1
H, d, J = 8.3 Hz), 8.18 (1H, d, J =
6.4 Hz), 8.22 (1H, d, J = 7.3H
z), 8.55 (1H, d, J = 6.4 Hz), 9.2
4 (1H, s)

Examples of the administration form of the compound of the present invention include oral administration such as tablets, capsules, granules, powders or syrups, and parenteral administration such as injections or suppositories. These formulations include excipients,
It is manufactured by a well-known method using additives such as a binder, a disintegrant, a lubricant, a stabilizer, and a flavoring agent. The amount used varies depending on symptoms, age, etc., but 0.01 to 20 mg / day
kg can be usually administered to an adult once a day or in several divided doses.

[0028]

EXAMPLES The present invention will now be described in more detail with reference to examples, but the present invention is not limited to these examples.

Examples 1-3 The compounds of the invention are shown by the following tests to have therapeutically valuable properties as ADM resistance-eliminating agents. The example numbers correspond to the compound numbers described above. [ADM resistance release test] The MTT assay for examining resistance release is Cancer Research, 47th.
Vol., 936-942 (1987). That is, human ovarian cancer cell lines NOS ₂ , NO
10,000 strains / m of S ₃ and each ADM resistant strain
It is suspended in a culture solution so as to have a volume of 1 and 200 μl is dispensed into a 96-well culture plate. ADM and the compounds of Example Nos. (1) to (3) are added to each well at a desired concentration, mixed, and cultured. After 4 days, MTT reagent is added to each well and the culture is continued for another 4 hours. The culture supernatant was removed, 200 μl of DMSO was added to each well, the cells were lysed by shaking, and the absorbance at 550 nm was measured.
The IC ₅₀ for DM was determined. The results are shown below (Table 2).

[0030]

[Table 2] Table 2 Example number NOS ₂ NOS ₂ AR NOS ₃ NOS ₃ AR control 0.086 5.800 0.090 1.700 (1) 0.069 0.547 0.033 0.250 (2) 0.072 4.210 0.045 1.570 (3) 0.063 0.448 0.025 0.177

[Toxicity] Oral administration of 500 mg / kg and 200 mg / k to fasted mice (5 mice per group) for 18 to 20 hours
Although acute symptoms were observed after intraperitoneal injection, no death was observed in any mouse.

Example 4 Formulation agents for the compounds of the present invention are shown below. 1. Tablets The following components can be prepared by known methods to give tablets. Ingredients Amount Compound (3) Phosphate 20 mg crystalline cellulose 20 mg Lactose 65 mg Corn starch 28.5mg Magnesium stearate 1.5mg calcium carboxymethylcellulose 5 mg ────────────── ───────────────── 140.0 mg

2. Sterile injectable solution The following ingredients are adjusted by a known method to make a total amount of 2 ml, and this is filled in an ampoule, sealed, and sterilized by heating to give a sterile injectable solution. Ingredients Amount Compound (3) phosphate 20 mg Sodium chloride 12 mg distilled water q.s. ───────────────────────────── ── Make the total volume 2 ml

[0034]

The usefulness of the compound of the present invention as an ADM resistance-eliminating agent is clear from Table 2 above.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Office reference number FI technical display location C07D 295/12 Z 401/12 243 8829-4C (A61K 31/70 31: 495) 9360-4C ( A61K 31/70 31:55) 9360-4C

Claims (1)

[Claims] 1. The following general formula (Formula 1): (In the formula, R ₁ is a 5-isoquinolyl group or a p-aminoethyloxyphenyl group, R ₂ and R ₃ are hydrogen or a methyl group, n is 2 or 3, and R ₄ is a benzyloxycarbonyl group or 3- A compound having a phenylpropionyl group) or a physiologically acceptable acid addition salt thereof as an active ingredient.