KR19980052668A - Novel camptothecin derivatives showing anticancer activity - Google Patents

Abstract

The present invention relates to a novel camptothecin derivative exhibiting anticancer activity.

[Formula 1]

In the above formula,

R ₁ is a hydrogen atom or a halogen atom;

R ₂ is hydrogen atom, halogen atom, hydroxy, amino, aniline, haloaniline, C ₁ to C ₇ alkylamino, C ₁ to C ₇ alkoxy, phenoxy, C ₁ to C ₇ aminoalkoxy, mercapto, C ₁ A C ₄ alkylsulphide, a C ₁ -C ₄ alkylsulfinyl or a C ₁ -C ₄ alkylsulfonyl group;

R ₃ is a hydrogen atom or —CH ₂ —Q wherein Q is a hydrogen atom, a halogen atom, hydroxy, piperazinyl, piperidinyl, 4-piperidinopiperidinyl, imidazolyl, pyrazolyl, 4 -(1-pyrrolidinyl) piperidinyl, 3-pyrroylyl, succinimido or phthalimido groups.

Description

Novel camptothecin derivatives showing anticancer activity

The present invention relates to a novel camptothecin derivative exhibiting anticancer activity.

In the above formula,

R ₁ is a hydrogen atom or a halogen atom;

R ₂ is hydrogen atom, halogen atom, hydroxy, amino, aniline, haloaniline, C ₁ to C ₇ alkylamino, C ₁ to C ₇ alkoxy, phenoxy, C ₁ to C ₇ aminoalkoxy, mercapto, C ₁ A C ₄ alkylthio, a C ₁ -C ₄ alkylsulfinyl or a C ₁ -C ₄ alkylsulfonyl group;

R ₃ is a hydrogen atom or —CH ₂ —Q wherein Q is a hydrogen atom, a halogen atom, hydroxy, piperazinyl, piperidinyl, 4-piperidinopiperidinyl, imidazolyl, pyrazolyl, 4 -(1-pyrrolidinyl) piperidinyl, 3-pyrroylyl, succinimido or phthalimido groups.

Camptothecin is a pentagonal alkaloid with conjugated quinoline, pyrroline, α-pyridone, and lactone ring, first extracted from Chinese tree ( C amptotheca acuminata Decsne.) By Wall et al. [ J Am. Chem. Soc. 88, 3888 (1966).

Camptothecin is also known to be the only compound that exhibits anticancer activity by inhibiting the action of topoisomerase I, one of the enzymes involved in cell growth. It is known that the anticancer activity of camptothecin is determined by the α-hydroxy lactone ring having the S configuration at the C-20 position, and it has been spotlighted as the most potent anticancer active substance due to its excellent effect. Is in progress.

However, camptothecin has a disadvantage in that it is very poor in solubility in water in formulating, and is very toxic and has limitations in application to the clinic. Therefore, there is an urgent need for the development of new camptothecin derivatives that can solve such problems by introducing new substituents that can increase solubility and reduce toxicity or by modifying the structure of the parent nucleus.

The present inventors have been made in view of the above demands to provide a novel camtosesin derivative having excellent solubility in water, easy to formulate, low toxicity, and excellent anticancer activity.

The present invention is characterized by the camtosesin derivative represented by the following formula (1).

Formula 1

In the above formula, R ₁ , R ₂ and R ₃ are each as defined above.

Referring to the present invention in more detail as follows.

The present invention relates to a novel camtosesin derivative having excellent anticancer activity, and among the derivatives represented by the general formula (1), particularly preferred compounds have the following substituents.

For example, R ₁ is preferably a hydrogen atom or a halogen atom. R ₂ represents a hydrogen atom, a halogen atom, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, amino, dimethylamino, dieethylamino, diisopropylamino, isopropylamino, cyclopropylamino, aniline , p-chloroaniline, dimethylaminoethoxy, aziridineethoxy, mercapto, methylthio, methylsulfinyl, mesyl. R ₃ is hydrogen or CH ₂ -Q, wherein Q is a halogen atom, hydroxy, piperazinyl, piperidinyl, 4-piperidinopiperidinyl, imidazolyl, pyrazolyl, 4- (1-pyrroli Dinyl) piperidinyl, 3-pyrroyl, succinimido or phthalimido.

The camtosesin derivative represented by Formula 1 of the present invention is a racemic or (S) -optical isomer.

As shown in the following Scheme 1, the camtosesin derivative represented by Formula 1 according to the present invention acidic 3-amino-2-pyrazinealdehyde derivative represented by the following formula (2) and the tricyclic compound represented by the following formula (3) Condensation reaction under conditions. The hydroxymethyl is then introduced to the C-7 position using a radical reaction [HETEROCYCLS, 1981, 16, 10, 1713], followed by the introduction of various R ₃ substituents.

R ₁ , R ₂ and R ₃ in the scheme are as defined above, respectively.

In the condensation reaction, when the (S) -3 bicyclic compound resolution (S) -array is resolved as a compound represented by Formula 3, a camtosesine derivative (1) of the S-array optical isomer is obtained, and is represented by Formula 3 When the racemic compound is used as the compound, racemic camptocecin derivative (1) is obtained. In addition, racemic camptothecin derivatives can only obtain (S) -optical isomers through resolution. The division used in the present invention is based on, for example, chromatography, recrystallization and the like as usual methods.

In addition, the 3-amino-2-pyrazinealdehyde derivative represented by the formula (2) used in the present invention can be easily prepared through general substitution, oxidation, and reduction from 3-aminopyrazinecarboxylate. The tricyclic compound represented by Chemical Formula 3 was synthesized in a racemic form by the Danishsky synthesis method [US Pat. No. 5,391,745], and only 20 (S) -optical isomer was obtained through conventional resolution. It may be.

As described above, the camtosesin derivative of the present invention was found to have excellent anticancer activity as a result of in vitro cytotoxic activity and in vivo anticancer activity measurement.

The present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1: 3-amino-2-pyrazinealdehyde

Methyl-3-amino-2-pyrazinecarboxylate (300 mg, 2 mmol) was dissolved in tetrahydrofuran (3 mL) and then 1.0 M of diisobutylaluminum hydride (DIBAL-H; 10; ML, 10 mmol) was added and stirred for 5 hours before methanol (2 mL) was added. After stirring at room temperature for 30 minutes, 1N HCl solution was added, followed by extraction with die ether, and silica gel column chromatography (elution solution: ethyl acetate / hexane = 1/2 by volume). As a result, 71.5 mg (yield 29.6%) of aldehyde compound and 99.9 mg (yield 33.0%) of ester compound were recovered.

¹ H-NMR (CDCl ₃ ), δ ppm: 10.02 (s, 1H), 8.21 to 8.05 (dd, 2H)

Example 2: 3-amino-5,6-dichloro-2-pyrazinealdehyde

Methyl-3-amino-5,6-dichloro-2-pyrazinecarboxylate (200 mg, 0.09 mmol) was dissolved in methylene chloride (5 ml), followed by 1.0 M of diisobutylaluminum hydride (1.0 M THF). , 2.7 mL, 0.27 mmol) was added and stirred at 0 ° C. for 5 hours before the addition of methanol. After stirring at room temperature for 30 minutes, 1N HCl solution was added, followed by extraction with die ether, and silica gel column chromatography (elution solution: ethyl acetate / hexane = 1/3 volume ratio) to give an alcohol compound 146 mg (yield 83.5%). Got.

¹ H-NMR (CDCl ₃ ), δ ppm: 5.20 (br.s, 2H), 4.71 (s, 2H)

The alcohol compound (146 mg, 0.75 mmol) obtained above was dissolved in methylene chloride (20 mL), followed by addition of pyridinium dichloromate (PDC; 566 mg, 1.5 mmol) and 4 ′ molecular sieve (900 mg). It stirred at 9 degreeC. Ethyl acetate (5 ml) was added, followed by filtration and silica gel column chromatography (eluent; ethyl acetate / hexane = 1/8, 1/6 by volume) to obtain 91 mg of the target aldehyde compound (yield 63.0%).

H-NMR (CDCl ₃ ), δ ppm: 9.94 (s, 1H)

Example 3: Preparation of Compound No. 1

The tricyclic compound (130 mg, 0.5 mmol) and 3-amino pyrazinealdehyde (180 mg, 1.5 mmol) were added to toluene (20 mL), refluxed for 30 minutes, and then catalytic amount of p-toluenesulfonic acid was added for 15 hours. It was refluxed using a Dean-Stark trap. Water was added, extraction was performed with methylene chloride, and silica gel column chromatography (eluent: 10% methanol / ethyl acetate) gave 80 mg (yield 46.3%) of the title compound.

¹ H-NMR (CDCl ₃ ), δ ppm: 9.15 to 9.11 (dd, 2H), 8.69 (s, 1H), 7.85 (s, 1H), 5.70 to 5.27 (dd, 2H), 5.40 (s, 2H) , 1.90-1.75 (q, 2H), 1.05-0.97 (t, 3H)

Example 4: Preparation of Compound No. 2

The compound prepared in Example 3 (Compound No. 1; 30 mg, 0.09 mmol) was dissolved in a mixed solvent of methanol (0.9 mL) and 50% concentrated sulfuric acid (1.4 mL), and FeSO ₄ (24 mg) was added thereto. 30% hydrogen peroxide (0.3 mL) was added slowly at 0 ° C. for 1 hour. After stirring for 8 hours while slowly raising the temperature to room temperature, water (10 mL) was added and filtered to obtain 18.5 mg (yield 56.7%) of the desired compound.

¹ H-NMR (CDCl ₃ ), δ ppm: 9.16 to 9.11 (dd, 2H), 7.84 (s, 1H), 5.71 to 5.27 (dd, 2H), 5.42 (s, 2H), 3.27 (s, 2H) , 1.89 to 1.74 (q, 2H), 1.03 to 0.96 (t, 3H)

Example 5: Preparation of Compound No. 5

The compound obtained in Example 4 (Compound No. 2; 20 mg, 0.05 mmol), succinimide (0.05 mL), triphenylphosphine (0.1 mmol) was dissolved in tetrahydrofuran (2 mL) under nitrogen, and then Ethyl azodicarboxylate (DEAD; 0.06 mmol) was added slowly at room temperature.

After stirring for 30 minutes, tetrahydrofuran was removed, and silica gel column chromatography (10% methanol / ethyl acetate) gave 16.2 mg of the target compound (yield 67%).

H-NMR (CDCl ₃ ), δ ppm: 9.17 to 9.09 (dd, 2H), 7.82 (s, 1H), 5.72 to 5.28 (dd, 2H), 5.41 (s, 2H), 5.19 (s, 2H), 2.65 (s, 4H), 1.90-1.77 (q, 2H), 1.02-0.96 (t, 3H)

Other camptothecin derivatives according to the present invention were prepared by the same preparation method as in Example with different benzopyran derivatives represented by the formula (2).

Some compounds prepared in the present invention are shown in the following Tables 1a, 1b, 1c and 1d.

Compound number R ₁ R ₂ R ₃ yield(%) ¹ H-NMR (CDC1 ₃ ), δ ppm One H H H 46.3 9.15-9.91 (dd, 2H), 8.69 (s, 1H), 7.85 (s, 1H), 5.70-5.27 (dd, 2H), 5.40 (s, 2H), 1.90-1.75 (q, 2H), 1.05- 0.97 (t, 3 H) 2 H H CH ₂ OH 56.7 9.16-9.91 (dd, 2H), 7.84 (s, 1H), 5.71-5.27 (dd, 2H), 5.42 (s, 2H), 3.27 (s, 2H), 1.89-1.74 (q, 2H), 1.03- 0.96 (t, 3 H) 3 H H CH ₂ Cl 92.0 9.14-9.10 (dd, 2H), 7.86 (s, 1H), 5.70-5.28 (dd, 2H), 5.39 (s, 2H), 4.05 (s, 2H), 1.88-1.73 (q, 2H), 1.02- 0.97 (t, 3 H) 4 H H CH ₂ Br 89.5 9.14-9.10 (dd, 2H), 7.84 (s, 1H), 5.68-5.27 (dd, 2H), 5.40 (s, 2H), 4.12 (s, 2H), 1.89-1.74 (q, 2H), 1.03- 0.97 (t, 3 H) 5 H H 67.0 9.17-9.09 (dd, 2H), 7.82 (s, 1H), 5.72-5.28 (dd, 2H), 5.41 (s, 2H), 5.19 (s, 2H), 2.65 (s, 4H), 1.90-1.77 ( q, 2H), 1.02-0.96 (t, 3H) 6 H H 70.0 9.17 to 9.10 (dd, 2H), 8.01 to 7.75 (m, 5H), 5.70 to 5.28 (dd, 2H), 5.41 (s, 2H), 5.01 (s, 2H), 1.91 to 1.78 (q, 2H), 1.02 to 0.97 (t, 3H) 7 H NH ₂ H 71.4 8.87 (s, 1H), 8.66 (s, 1H), 7.82 (s, 1H), 5.73-5.30 (dd, 2H), 5.43 (s, 2H), 1.92-1.77 (q, 2H), 1.04-0.96 ( t, 3H) 8 Cl Cl H 60.3 8.58 (s, 1H), 7.80 (s, 1H), 5.77-5.24 (dd, 2H), 5.38 (s, 2H), 1.88-1.74 (q, 2H), 1.04-1.00 (t, 3H)

Compound number R ₁ R ₂ R ₃ yield(%) ¹ H-NMR (CDC1 ₃ ), δ ppm 9 Cl Cl CH ₂ OH 53.2 7.82 (s, 1H), 5.75 to 5.33 (dd, 2H), 5.36 (s, 2H), 3.51 (s, 2H), 1.87 to 1.74 (q, 2H), 1.04 to 0.98 (t, 3H) 10 Cl Cl CH ₂ Cl 89.1 7.79 (s, 1H), 5.76 to 5.33 (dd, 2H), 5.39 (s, 2H), 4.10 (s, 2H), 1.88 to 1.74 (q, 2H), 1.03 to 0.99 (t, 3H) 11 Cl Cl CH ₂ Br 90.8 7.78 (s, 1H), 5.75 to 5.22 (dd, 2H), 5.37 (s, 2H), 4.17 (s, 2H), 1.88 to 1.73 (q, 2H), 1.02 to 0.99 (t, 3H) 12 Cl Cl 72.1 7.78 (s, 1H), 5.79 to 5.26 (dd, 2H), 5.41 (s, 2H), 5.18 (s, 2H), 2.61 (s, 4H), 1.91 to 1.77 (q, 2H), 1.05 to 1.01 ( t, 3H) 13 Cl NH ₂ H 65.7 8.43 (s, 1H), 7.75 (s, 1H), 5.74-5.23 (dd, 2H), 5.27 (s, 2H), 1.85-1.73 (q, 2H), 1.02-0.93 (t, 3H) 14 Cl N (CH ₃ ) ₂ H 61.5 8.40 (s, 1H), 7.74 (s, 1H), 5.55 (dd, 2H), 5.25 (s, 2H), 2.90 (s, 6H), 1.76 (q, 2H), 0.98 (t, 3H) 15 Cl OCH ₃ H 65.2 8.56 (s, 1H), 7.75 (s, 1H), 5.76 to 5.21 (dd, 2H), 5.27 (s, 2H), 4.02 (s, 3H), 1.90 to 1.77 (q, 2H), 1.10 to 0.98 ( t, 3H) 16 Cl OEt H 70.5 8.52 (s, 1H), 7.72 (s, 1H), 5.45 (dd, 2H), 5.26 (s, 2H), 4.09 (q, 2H), 1.85 (q, 2H), 1.45 (t, 3H), 1.03 (t, 3H)

Compound number R ₁ R ₂ R ₃ yield(%) ¹ H-NMR (CDC1 ₃ ), δ ppm 17 Cl OH H 73.8 8.52 (s, 1H), 7.70 (s, 1H), 5.51 (dd, 2H), 5.30 (s, 2H), 1.82 (q, 2H), 1.01 (t, 3H) 18 Cl OCH ₂ CH ₂ -N (CH ₃ ) ₂ H 60.4 8.54 (s, 1H), 7.72 (s, 1H), 5.50 (dd, 2H), 5.31 (s, 2H), 3.52 (t, 2H), 2.92 (t, 2H), 2.35 (s, 6H), 1.79 (q, 2H), 1.02 (t, 3H) 19 Cl SH H 69.7 8.45 (s, 1H), 7.61 (s, 1H), 5.51 (dd, 2H), 5.30 (s, 2H), 1.65 (q, 2H), 0.99 (t, 3H) 20 Cl SCH ₃ H 62.5 8.47 (s, 1H), 7.65 (s, 1H), 5.45 (dd, 2H), 5.32 (s, 2H), 2.61 (t, 3H), 1.79 (q, 2H), 1.02 (t, 3H) 21 Cl SOCH ₃ H 52.6 8.50 (s, 1H), 7.69 (s, 1H), 5.52 (dd, 2H), 5.27 (s, 2H), 2.76 (t, 3H), 1.67 (q, 2H), 1.00 (t, 3H) 22 Cl SO ₂ CH ₃ H 57.7 8.48 (s, 1H), 7.66 (s, 1H), 5.51 (dd, 2H), 5.30 (s, 2H), 3.02 (t, 3H), 1.71 (q, 2H), 1.01 (t, 3H) 23 Br H H 59.1 9.10 (s, 1H), 8.55 (s, 1H), 7.77 (s, 1H), 5.77-5.21 (dd, 2H), 5.39 (s, 2H), 1.87-1.75 (q, 2H), 1.08-0.97 ( t, 3H) 24 Br H CH ₂ OH 56.9 9.07 (s, 1H), 7.79 (s, 1H), 5.52 (dd, 2H), 5.27 (s, 2H), 3.40 (s, 2H), 1.80 (q, 2H), 1.02 (t, 3H) 25 Br H CH ₂ Cl 87.3 9.08 (s, 1H), 7.80 (s, 1H), 5.49 (dd, 2H), 5.35 (s, 2H), 4.05 (s, 2H), 1.79 (q, 2H), 1.01 (t, 3H)

Compound number R ₁ R ₂ R ₃ yield(%) ¹ H-NMR (CDC1 ₃ ), δ ppm 26 Br H 68.1 9.10 (s, 1H), 7.77 (s, 1H), 5.55 (dd, 2H), 5.30 (s, 2H), 5.09 (s, 2H), 2.57 (s, 4H), 1.69 (q, 2H), 1.00 (t, 3H) 27 I NH ₂ H 59.3 8.59 (s, 1H), 7.80 (s, 1H), 5.56 (dd, 2H), 5.32 (s, 2H), 1.80 (q, 2H), 1.03 (t, 3H) 28 H H 71.5 9.12 (dd, 2H), 7.80 (s, 1H), 7.51 (s, 1H), 7.02 (s, 1H), 6.95 (s, 1H), 5.56 (dd, 2H), 5.41 (s, 2H), 5.22 (s, 2H), 1.80 (m, 2H), 0.97 (t, 3H) 29 H H 68.0 8.80 (s, 1H), 8.59 (s, 1H), 7.79 (s, 1H), 7.30 ~ 6.87 (m, 5H), 5.56 (dd, 2H), 5.27 (s, 2H), 1.86 (q, 2H) , 0.99 (t, 3H)

Experimental Example 1 In Vitro Cytotoxicity

Cytotoxicity was measured by Carmichael's MTT method. Next, the cell lines shown in Table 2 were cultured to an exponential growth stage to a constant concentration in a T-25 incubator to make a single cell suspension. Incubated for 24 hours in a CO ₂ incubator. The drug of each concentration was dissolved in the medium, and 100 l of each was added, followed by further incubation for 3-5 days. After incubation, 20 ml of MTT solution was added to each well and incubated for 4 hours. After dissolving the dye with dimethyl sulfoxide (DMSO), the absorbance was measured at 570 nm. Cytotoxicity was evaluated by measuring the survival rate by substituting the measured absorbance into the following Equation 1.

In this case, 1 column (8 wells) was used under the same conditions for one concentration group, and the final concentration of the sample was 300, 100, and 30 g / ml per well, respectively. As a control group, only 100 L of PBS was added instead of the drug in one column to make a 100% survival group. Only 100 l of cell-free medium was added to the standard sample used for absorbance measurement, and 100 l of PBS or drug was added.

In addition, the cytotoxicity of the selection material was expressed as a 50% inhibitory concentration (IC ₅₀ ) that can inhibit cell proliferation by 50%.

50% inhibitory concentration (IC ₅₀ ; μg / ml) Medication SNU-5 SNU-398 SNU-16 SNU-C5 P388 Camptosesin 4.43 × 10 ^-6 5.77 × 10 ^-5 1.03 × 10 ^-5 6.47 × 10 ^-5 1.82 × 10 ^-3 Compound number 1 (racemic) 4.04 1.70 × 10 ^-2 8.18 × 10 ^-2 6.09 × 10 ^-3 1.87 × 10 ^-2 Compound number 1 ((S) -isomer) 1.13 6.97 × 10 ^-3 5.24 × 10 ^-2 2.58 × 10 ^-3 7.21 × 10 ^-3 Compound number 8 ((S) -isomer) 6.57 4.30 × 10 ^-2 9.86 × 10 ^-2 9.58 × 10 ^-3 5.32 × 10 ^-2 SNU-5: Domestic gastric cancer cell line SNU-398: Domestic liver cancer cell line SNU-C5: Domestic colon cancer cell line SNU-16: Domestic gastric cancer cell line P388: Mouse leukemia cell line

Experimental Example 2 Tumor Colonization Rate

The Salmon tumor tumor formation performance test, which can compensate for in vitro cytotoxicity, was modified to compare the inhibition rate, 50% inhibitory concentration (IC ₅₀ ), and to compare the performance.

A 1-fold agarose (agarose) and 20% fetal bovine serum containing a 2-fold concentrated RPMI1640 medium in a 1: 1 mixture and dispensed on a 35 mm plate to solidify at room temperature to prepare a base layer of agar, and a constant concentration of cell lines and drugs Was reacted for 1 hour or several hours in PRMI1640 medium containing 10% fetal bovine serum, washed three times with RPMI1640, and 1 ml aliquots were prepared on the previously prepared agar base layer and cultured in a 5% CO ₂ , 37 ° C. incubator. Whether or not was observed by an inverted microscope.

Experimental Example 3: In vivo anticancer activity

In vivo anticancer activity was evaluated by investigating the survival rate by administering the drug to mice transplanted with P388 (mouse leukemia cell line).

Anticancer Effect on P388 Tumors

As a model for measuring anti-cancer activity in vivo, 1 × 10 ⁶ P388 leukemia cells were transplanted into BDF1 (7 week old female, 5 group 1) mice, and the compounds of the present invention were used as positive controls 24 hours after transplantation. Tomycin was intraperitoneally administered once a day for 9 days a day to check the survival rate, and a saline-administered group was used as a control group. The results are shown in Table 3 below.

Medication P388 cells Dose (μg / horse) Average survival (days) % T / C Mitomycin 40 26 137 Compound number 1 (racemic) 40 26 137 Compound number 1 ((S) -isomer) 40 30 158 Compound number 8 ((S) -isomer) 40 24 126 Control - 19 100

According to the results of the in vivo experiment of Table 3, in the case of the well-known anticancer drug mitomycin, the% T / C value for P388 cells was 137. On the other hand, the novel derivatives according to the present invention showed a very high value between 140 and 160, thereby indicating that the anticancer activity of the compound of the present invention is very excellent.

The camtosesin derivative represented by Formula 1 according to the present invention is very useful as an anticancer agent because of its excellent anticancer activity.

Claims (3)

Kamtosesin derivative represented by the following formula (1). Formula 1 In the above formula, R ₁ is a hydrogen atom or a halogen atom; R ₂ is hydrogen atom, halogen atom, hydroxy, amino, aniline, haloaniline, C ₁ to C ₇ alkylamino, C ₁ to C ₇ alkoxy, phenoxy, C ₁ to C ₇ aminoalkoxy, mercapto, C ₁ A C ₄ alkylthio, a C ₁ -C ₄ alkylsulfinyl or a C ₁ -C ₄ alkylsulfonyl group; R ₃ is a hydrogen atom or —CH ₂ —Q wherein Q is a hydrogen atom, a halogen atom, hydroxy, piperazinyl, piperidinyl, 4-piperidinopiperidinyl, imidazolyl, pyrazolyl, 4 -(1-pyrrolidinyl) piperidinyl, 3-pyrroylyl, succinimido or phthalimido groups. The method of claim 1, R ₁ is a hydrogen atom or a halogen atom, R ₂ represents a hydrogen atom, a halogen atom, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, phenoxy, amino, dimethylamino, diethamino, diisopropylamino, isopropylamino, cyclo Propylamino, aniline, p-chloroaniline, dimethylaminoethoxy, aziridineethoxy, mercapto, methylthio, methylsulfinyl or mesyl groups; R ₃ is a hydrogen atom or -CH ₂ -Q wherein Q is a halogen atom, hydroxy, piperazine, piperidine, 4-piperidinopiperidine, imidazole, pyrazole, 4- (1-pipe Olidinyl) piperidine, 3-pyrroline, succinimido or phthalimido groups. The method of claim 2, R ₁ is a hydrogen atom or a halogen atom, R ₂ is hydrogen atom, chlorine atom, hydroxy, methoxy, ethoxy, amino, dimethylamino, aniline, methylaminoethoxy, mercapto, methylthio, methylsulfinyl, mesyl; R ₃ is a hydrogen atom or —CH ₂ —Q, wherein Q is a chlorine atom, iodo atom, hydroxy, imidazole, succinimido or phthalimido group.