KR100265033B1 - Hexacyclic camptothecin analogs and process for prsparing the same - Google Patents

Abstract

The present invention relates to a novel hexacyclic camptothecin derivative represented by the following formula (1) having anticancer activity, a pharmaceutically acceptable salt thereof, a preparation method thereof, and a pharmaceutical composition containing the same as an active ingredient.

In Formula 1 above:

R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or a C ₃ -C ₆ cycloalkyl group;

R ² represents a cyano group or an aminoalkyl group of C ₁ ~C ₇ a represents a hydrogen _{atom, C 1 ~C 6, C 1} ~C 6.

Description

Hexacyclic camptothecin derivatives and preparation methods thereof

The present invention relates to a novel hexacyclic camptothecin derivative represented by the following formula (1) having anticancer activity, a pharmaceutically acceptable salt thereof, a preparation method thereof, and a pharmaceutical composition containing the same as an active ingredient.

Formula 1

In Formula 1 above:

R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or a C ₃ -C ₆ cycloalkyl group;

R ² represents a cyano group or an aminoalkyl group of C ₁ ~C ₇ a represents a hydrogen _{atom, C 1 ~C 6, C 1} ~C 6.

In general, camptothecin and its derivatives are well known to have anticancer activity due to their inhibitory activity against topoisomerase I, which is involved in the rotation and relaxation of DNA replication and transcription [Beppino et al. , Science, 1989, 246, 1046; Kingsbury et al., J. Med. Chem., 1991, 34, 98]. Camptocecin and its derivatives have been reported to exhibit better anticancer activity in lactone structures than in carboxylate structures in in vitro and in vivo conditions [Wani et al., J. Med. Chem., 1987, 30, 1774; Wani et al., J. Med. Chem., 1987, 30, 2317; Jaxel et al., Cancer Res., 1989, 49, 5077; Slichenmeyer et al., J. Natl. Cancer Inst., 1993, 85, 271]. This is because the ring-closed α-hydroxy δ-lactone ring portion effectively works with topoisomerase I [Hertzberg et al., J. Med. Chem., 1989, 32, 715, and due to the structural specificity of having anticancer activity in in vivo conditions [Giovanella et al., Cancer Res., 1991, 51, 3052]. Recently reported, in the presence of physiologically appropriate levels of human albumin, the biologically active camptothecin has a very short half-life of about 12 minutes, and more than 99% of the camtosesin is 2 hours after administration to human plasma. Converted to carboxylate, which is biologically inactive and toxic [Burke, TG and Mi, Z., J. Med. Chem., 1994, 37, 40]. Therefore, there is an urgent need for novel camptothecin derivatives having high anticancer activity and stability in the presence of human serum albumin. Burke's and Mi's work is to modify the C7- and C9-positions of the quinoline nucleotides, as in CPT-11, topotecan and SN-38, to promote stability in the presence of human serum albumin.

In addition, the condensation of five-membered heterocycles at the C9- and C10-positions of the camptothecin ring has recently resulted in a high anticancer effect due to the inhibitory action on topoisomerase I. 96/38449; WO 97/25332].

Therefore, the inventors of the present invention synthesized a novel compound condensed with a 6-membered heterocycle at the C9- and C10-positions of the camptothecin ring, and the novel compound of the present invention has high anticancer activity to satisfy the above requirements. The present invention was completed by knowing.

Accordingly, an object of the present invention is to provide a novel hexacyclic camptothecin derivative represented by the formula (1) and a pharmaceutically acceptable salt thereof. In addition, the present invention has another object to provide a method for producing a compound represented by the formula (1). Another object of the present invention is to provide an anticancer drug composition containing the compound represented by the formula (1) as an active ingredient.

The present invention is characterized by the hexacyclic camptothecin derivative represented by the following formula (1) and its pharmaceutically acceptable salts.

Formula 1

In Formula 1 above:

R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or a C ₃ -C ₆ cycloalkyl group;

R ² represents a cyano group or an aminoalkyl group of C ₁ ~C ₇ a represents a hydrogen _{atom, C 1 ~C 6, C 1} ~C 6.

Referring to the present invention in more detail as follows.

In the hexacyclic camptothecin derivative represented by Chemical Formula 1 according to the present invention, the following is preferable. R ¹ is a hydrogen atom; Methyl, ethyl, n- propyl, and i- propyl group of the C ₁ ~C ₆ containing group; Comprising a cyclo propyl, cyclo-butyl group, and a cyclo-pentyl group when the cycle of the C ₃ ~C _6. And R ² is a hydrogen atom; Comprising a methyl group, an ethyl group, a propyl group and a cyclo propyl group of the C ₁ ~C _6; C ₁ to C ₆ cyanoalkyl group containing a cyanomethyl group, a cyanoethyl group and a cyanopropyl group; Or a C ₁ to C ₇ aminoalkyl group containing an aminomethyl group, an aminoethyl group and an aminopropyl group.

Particularly preferred hexacyclic camptothecin derivatives represented by Formula 1 are as follows:

(9S) -9-ethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6,7] indoridge No [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 1),

(9S) -9,16-diethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6,7 ] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 2),

(9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-methyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6, 7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 3),

(9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-ethyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6, 7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 4),

(9S) -9-Ethyl-2,3-dihydro-9-hydroxy-1-cyanomethyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 5),

(9S) -9-ethyl-2,3-dihydro-9-hydroxy-1- (2-aminoethyl) -12H-1,4-oxazino [3,2-f] pyrano [3 ', 4 ': 6,7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione acetate salt (Compound No. 6),

(9S) -9,16-diethyl-2,3-dihydro-9-hydroxy-1-methyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4' : 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 7).

In addition, the hexacyclic camptothecin derivative represented by Chemical Formula 1 according to the present invention may form a pharmaceutically acceptable acid addition salt according to a conventional method in the art, for example, dissolving a compound in an appropriately selected solvent. It is prepared by treating with an excess of inorganic or organic acid. Pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid or nitric acid; And organic acid salts such as acetic acid, methanesulfonic acid, citric acid, fumaric acid, maleic acid, ascorbic acid, succinic acid, tartaric acid, benzenesulfonic acid and the like.

The novel hexacyclic camptothecin derivatives represented by Formula 1 according to the present invention and pharmaceutically acceptable salts thereof have high anticancer activity against various types of solid cancer and leukemia. Therefore, the present invention includes a pharmaceutical composition containing an effective amount of at least one or more compounds selected from the derivatives represented by the formula (1). These pharmaceutical compositions are formulated as conventional formulations in the pharmaceutical art by adding conventional nontoxic pharmaceutically acceptable carriers, adjuvant and excipients to the compound represented by Formula 1 (e.g., oral, parenteral, intravenous, Intraperitoneal, subcutaneous, topical) for administration in solid or liquid form. In particular, it is preferably administered orally and intravenously.

In clinical use of the pharmaceutical composition of the present invention, it is combined with a conventional carrier in the pharmaceutical field, and oral administration such as tablets, capsules, troches, elixirs, solutions, suspensions, etc. Preparations for use; Injectable preparations, such as injectable solutions, injectable lyophilized powders or suspensions; It can be formulated into various forms such as topical preparations such as ointments, creams, liquids and the like. Carriers that can be used in the pharmaceutical composition of the present invention should be non-toxic as is commonly applied in the pharmaceutical field.

In the case of preparations for oral administration, binders such as microcrystalline cellulose, gumtragacanth or gelatin; Disintegrants such as alginic acid, Prinogel ™ and corn starch; Lubricants such as magnesium stearate and Steotes ™; Glidant such as colloidal silica; Sweeteners such as sugar, saccharin and / or seasonings such as peppermint, methyl salicylate, orange flavor and the like and the like can be applied. When the unit dosage form is a capsule, in addition to the additives described above may include a liquid carrier such as fatty oil. Other forms of unit dosage form may include materials that may improve the physical form of the dosage unit, such as coatings. Thus, in the case of tablets or pills, it may be coated with sugar shellac or coated with other enteric skin. In the case of syrup, in addition to the active compound, it may contain sugars, traditional preservatives, dyes, colorants, seasonings and the like as a sweetener.

Parenteral formulations can be used by preparing the active ingredient in solution or as a lyophilized powder or suspension. Ingredients that may be included in such solutions or lyophilized powders or suspensions include sterile diluents such as injectables, saline, hardened oils, polyethylene glycols, glycerin, propylene glycol, or other synthetic solvents; Antibacterial agents such as benzyl alcohol or methylparabens; Antioxidants such as ascorbic acid or sodium bisulfite; Chelating agents such as ethylene diaminetetraacetic acid (EDTA); Buffers such as acetates, citrate or phosphates; Reagents that control tonicity include sodium chloride, dextrose, mannose, and the like. Parenteral preparations may be used in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

The dosage of the new hexacyclic camptothecin derivative represented by Formula 1 according to the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient. The dose is 0.01 to 30 mg / kg body weight, preferably 0.02 to 10 mg / kg body weight, and is determined by a doctor or pharmacist at a time interval of one day, one week, two weeks, three weeks, or four. It may be administered once or several times per week.

In addition, the present invention includes a method for preparing a hexacyclic camptothecin derivative, which is briefly shown in Scheme 1 below.

In Scheme 1: R ¹ and R ² are as defined above, respectively.

10-Hydroxy- (20S) -camptothecin or 7-ethyl-10-hydroxy- (20S) -camptothecin, which is used as a raw material in the preparation method according to the present invention, is a known compound, prepared from natural products or known It can be obtained by the semi-synthesis method or the total synthesis method which is a method [International Patent Publication WO 92/05785; United States Patent No. 5,468,754.

The brominated compound represented by Formula 3 may be selected from 10-hydroxy- (20S) -camptothecin or 7-ethyl-10-hydroxy- (20S) -camptothecin represented by Formula 2 in the presence of a suitable solvent and a suitable base. It can manufacture by reaction with, 2-dibromoethane. Suitable solvents that may be used at this time are N, N-dimethylformamide (DMF), dimethyl acetamide (DMA), dimethyl sulfoxide (DMSO), acetone, hexamethylphosphoamide (HMPA) and 1-methyl-2- Sole or mixed solvent selected from pyrrolidinone (NMP). Suitable bases are alkali or alkaline earth metal salts, for example K ₂ CO ₃ , Na ₂ CO ₃ , Cs ₂ CO ₃ or NaH and the like. Typical reaction temperatures range from 0 ° C. to 100 ° C. and reaction times range from 30 minutes to 1 day.

The nitro compound represented by Chemical Formula 4 may be prepared by nitrating the brominated compound represented by Chemical Formula 3. The nitration reaction can be carried out using concentrated sulfuric acid and concentrated nitric acid, or using only nitric acid. Or nitration reactions by the method described in J. March, “Advanced Organic Chemistry” 3rd Edition p.468, for example N ₂ O ₅ , CH ₃ NO ₃ and BF ₃ , NaNO ₂ and CF ₃ COOH, N ₂ O ₄ and the like is reacted for several minutes to 1 day in the temperature range -20 ℃ to 40 ℃.

The amino compound represented by Chemical Formula 5 is -20 using a nitro compound represented by Chemical Formula 4 using a reducing metal or metal salt (for example, SnCl ₂ , Zn or Fe) and a suitable solvent (for example, acetic acid, hydrochloric acid, etc.). It can be prepared by reacting for several minutes to several days in the temperature range of ℃ to 60 ℃.

The amino compound represented by the formula (5) prepared by the above preparation method may be converted into a compound represented by the formula (1) wherein R ² is a hydrogen atom by using an appropriate solvent or an appropriate solvent and an appropriate organic or inorganic base. Can be. At this time, suitable solvents are single or mixed solvents selected from N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), acetone, acetonitrile and dioxane. Suitable organic bases include trialkylamines (e.g. triethylamine or diisopropylethylamine), or heteroaromatic amines (for example pyridine or 2,6- (C ₁ ~C ₆ alkyl) pyridine, and the like) to be used have. Suitable inorganic bases are alkali or alkaline earth metal salts such as NaHCO ₃ , Na ₂ CO ₃ or K ₂ CO ₃ and the like. Further, in the cyclization reaction according to the present invention, a good yield may be obtained by adding an alkali metal iodide salt or an alkali earth iodide salt, for example, NaI. Typical reaction temperatures range from 0 ° C. to 200 ° C., and reaction times range from 5 minutes to 3 days.

In addition, in the case of the compound represented by the formula (1) wherein R ² is an alkyl group or a cyanoalkyl group, the compound wherein R ² is a hydrogen atom prepared above may be prepared by reacting with a suitable alkyl halide or cyanoalkyl halide in a suitable solvent. This reaction may be carried out without using a base, and may also be carried out in the presence of a suitable inorganic or organic base. As a reaction solvent, a conventional organic solvent is used, and preferably N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), hexamethylphosphoamide (HMPA), 1-methyl-2-pyrroli It is to use a single or mixed solvent selected from dinon (NMP) and dioxane. Suitable alkyl halide compounds include C ₁ -C ₆ alkyl halides comprising methane iodide (MeI), ethane iodide (EtI), methane bromide (MeBr), propane iodide (PrI), and suitable cyanoalkyl halide compounds. Uses C ₁ to C ₆ cyanoalkyl halides including cyanomethane bromide (BrCH ₂ CN), cyanoethane bromide (Br (CH ₂ ) ₂ CN), and cyanomide iodide (ICH ₂ CN) . Suitable inorganic or organic bases include the bases mentioned in the above cyclization reactions. The reaction temperature is typically 0 ° C to 200 ° C, the reaction time is about 10 minutes to 3 days, and preferably 1 hour to 24 hours.

In addition, in the case of the compound represented by the formula (1) wherein R ² is an aminoalkyl group, the compound represented by the formula (1) wherein R ² is a C ₁ to C ₆ cyanoalkyl group in the presence of a solvent containing a suitable acid and a suitable catalyst It can be prepared by a reduction reaction using molecular hydrogen (H ₂ ). Suitable acids are those of commonly used organic or inorganic acids, including acetic acid, hydrochloric acid, sulfuric acid or phosphoric acid. Suitable solvents are C ₁ -C ₄ alcohols (for example methanol, ethanol or protanol), chloroform, water, acetic acid or mixed solvents thereof. Suitable catalysts include transition metals such as palladium (Pd), platinum (Pt), nickel (Ni) or mixed metals thereof. Typically the reaction temperature is in the range of 0 ° C to 100 ° C, the reaction pressure is 1 atm to 100 atm, and the reaction time is about 1 hour to 3 days. Under preferred conditions, an acetic acid solvent is used under a Raney-Ni catalyst and hydrogen molecules (H ₂ ) are reduced at a pressure of 2 atm at 25 ° C.

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

Preparation Example 1 Preparation of 10- (2-bromoethoxy)-(20S) -chemtothecin (Formula 3, R ¹ = H)

To dry N, N-dimethylformamide (DMF; 800 mL) 10-hydroxy- (20S) -chemtocecin (30 g, 82.3 mmol), anhydrous 1,2-dibromoethane (310 g, 1.648 mol) , 142 mL) and anhydrous potassium carbonate (K ₂ CO ₃ ; 56.9 g, 412 mmol) were added thereto, and the mixture was stirred vigorously for 10 hours at 80 ° C. under a nitrogen stream. The reaction mixture was filtered through celite, the filtrate was washed with DMF, the filtrate and the resulting solution were collected and dried under reduced pressure. The residue was dissolved in 10% CH ₃ OH / CHCl ₃ solution (1.5 L), insoluble material was filtered off and the filtrate was concentrated under reduced pressure. The concentrate was adsorbed onto silica gel, placed on top of the silica gel column, and purified by MPLC (eluent: 3% CH ₃ OH / CHCl ₃ → 5% CH ₃ OH / CHCl ₃ ) to give 23.55 g (yield 61%) of the title compound. Obtained, which was recrystallized from CH ₃ OH / CHCl ₃ solution.

mp: 237-238 ℃

IR (neat): 3334, 1752 (lactone), 1660 (pyridone), 1603, 1556, 1505, 1239, 1156 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.99 (t, J = 7.2 Hz, 3H, H-18), 1.87 (m, 2H, H-19), 3.92 (t, J = 5.5 Hz, 2H, CH ₂ Br), 4.53 (t, J = 5.5 Hz, 2H, CH ₂ CH ₂ Br), 5.28 (s, 2H, H-5), 5.42 (s, 2H, H-17), 6.49 (br s, 1H, OH), 7.30 (s, 1H, H-14), 7.53 to 7.59 (m, 2H, H-9 and H-11), 8.10 (d, J = 9.0 Hz, 1H, H-12), 8.55 (s, 1H, H-7).

Preparation Example 2 Preparation of 10- (2-Bromoethoxy) -7-ethyl- (20S) -chemtothecin (Formula 3, R ¹ = CH ₂ CH ₃ )

In dried DMF (200 mL) 7-ethyl-10-hydroxy- (20S) -chemtocecin (7.87 g, 20.1 mmol), anhydrous 1,2-dibromoethane (75.30 g, 406 mmol, 35 mL) And anhydrous K ₂ CO ₃ (13.82 g, 100 mmol) were added and stirred vigorously for 10 hours at 80 ° C. under a nitrogen stream. The reaction mixture was filtered through celite, the filtrate was washed with DMF, the filtrate and the resulting solution were collected and dried under reduced pressure. The residue was dissolved in 10% CH ₃ OH / CHCl ₃ solution (150 mL), the insoluble material was filtered off and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel-filled MPLC (eluent: 1% CH ₃ OH / CHCl ₃ → 3% CH ₃ OH / CHCl ₃ ) to give 5.79 g (yield 58%) of the title compound, which was CH ₃ OH / CHCl _3. Recrystallized from solution.

mp: 236.9-237.2 ° C

IR (neat): 3326, 1755 (lactone), 1658 (pyridone), 1602, 1516, 1254 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.99 (t, J = 7.4 Hz, 3H, H-18), 1.30 (t, J = 7.5 Hz, 3H, CH ₂ CH ₃ ), 1.87 (m, 2H , H-19), 3.17 (q, J = 7.5 Hz, 2H, CH ₂ CH ₃ ), 3.93 (t, J = 5.3 Hz, 2H, CH ₂ Br), 4.58 (t, J = 5.3 Hz, 2H, CH ₂ CH ₂ Br), 5.25 (s, 2H, H-5), 5.42 (s, 2H, H-17), 7.26 (s, 1H, H-14), 7.50 (s, 1H, H-9) 7.52 (d, J = 9.0 Hz, 1H, H-11), 8.06 (d, J = 9.0 Hz, 1H, H-12).

Preparation Example 3 Preparation of 10- (2-bromoethoxy) -9-nitro- (20S) -chemtocecin (Formula 4, R ¹ = H)

10- (2-bromoethoxy)-(20S) -chemtocecin (12.55 g, 26.6 mmol) was added in small portions over 1 hour with stirring of concentrated sulfuric acid (175 mL) at 0 ° C. After the reaction solution was cooled to -10 ° C, a mixed solution of concentrated sulfuric acid (5 mL) and fuming nitric acid (5 mL) was slowly injected for 30 minutes while cooling to -10 ° C. The reaction mixture was warmed to 0 ° C., stirred for an additional 1 hour, and then poured slowly onto ice (800 g). The yellow precipitate was filtered off and washed sequentially with water, cold ethanol and diethyl ether. The aqueous filtrate was refiltered through celite and the celite filtercake was extracted with 30% CH ₃ OH / CHCl ₃ solution (500 mL). The organic solvent was distilled off under reduced pressure to obtain a yellow solid. Ethanol was used to obtain 11.68 g (yield 85%) of the title compound as a yellow solid powder, which was recrystallized from a CH ₃ OH / CHCl ₃ solution.

mp: 251.5 ° C (dec)

IR (neat): 3309, 1751 (lactone), 1656 (pyridone), 1598, 1531, 1257, 1166 cm ^-1

_{^{1 H NMR (DMSO-d 6}} ): δ0.88 (t, J = 7.4Hz, 3H, H-18), 1.87 (m, 2H, H-19), 3.86 (t, J = 5.4Hz, 2H, CH ₂ Br), 4.75 (t, J = 5.4 Hz, 2H, CH ₂ CH ₂ Br), 5.27 (s, 2H, H-5), 5.43 (s, 2H, H-17), 6.53 (br s, 1H, OH), 7.34 (s, 1H, H-14), 8.04 (d, J = 9.6 Hz, 1H, H-11), 8.43 (d, J = 9.6 Hz, 1H, H-12), 8.47 ( s, 1H, H-7).

Preparation Example 4 Preparation of 10- (2-bromoethoxy) -7-ethyl-9-nitro- (20S) -chemtocecin (Formula 4, R ¹ = CH ₂ CH ₃ )

10- (2-bromoethoxy) -7-ethyl- (20S) -chemtothecin (8.60 g, 17.2 mmol) was added in small portions over 20 minutes with stirring of concentrated sulfuric acid (175 mL) at 0 ° C. After stirring for 30 minutes and cooling the reaction solution to -10 ℃, a mixture of concentrated sulfuric acid (3.2 mL) and fuming nitric acid (32 mL) was slowly injected for 10 minutes while cooling to -10 ℃. The reaction mixture was warmed to 0 ° C., stirred for an additional 1 hour, and then poured slowly onto ice (1 kg). The yellow precipitate was filtered off and washed sequentially with water, cold ethanol and diethyl ether. The aqueous filtrate was extracted with 10% CH ₃ OH / CHCl ₃ solution (300 mL × 2). The filtered precipitate was dissolved in 10% CH ₃ OH / CHCl ₃ solution (700 mL), and then the organic layer solution was passed through a short silica gel column. Sikyeotgo reduced pressure to evaporate the organic solvent until a solid was generated, were obtained concentrate is scored diethyl ether the desired compound 8.58 g (92% yield) of a yellow solid was pulverized and filtered through a (200 ㎖), which CH ₃ Recrystallized from OH / CHCl ₃ solution.

mp: 268 ° C (dec)

IR (neat): 3236, 1751 (lactone), 1659 (pyridone), 1597, 1538, 1263, 1162 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.98 (t, J = 7.4 Hz, 3H, H-18), 1.21 (t, J = 7.4 Hz, 3H, CH ₂ CH ₃ ), 1.88 (m, 2H , H-19), 2.88 (q, J = 7.4 Hz, 2H, CH ₂ CH ₃ ), 3.83 (t, J = 5.3 Hz, 2H, CH ₂ Br), 4.72 (t, J = 5.3 Hz, 2H, CH ₂ CH ₂ Br), 5.34 (s, 2H, H-5), 5.44 (s, 2H, H-17), 7.31 (s, 1H, H-14), 8.03 (d, J = 9.3 Hz, 1H , H-11), 8.40 (d, J = 9.3 Hz, 1H, H-12).

Preparation Example 5 Preparation of 9-amino-10- (2-bromoethoxy)-(20S) -chemtocecin (Formula 5, R ¹ = H)

10- (2-bromoethoxy) -9-nitro- (20S) -chemtothecin (815 mg, 1.579 mmol) was added in small portions over 10 minutes with stirring of concentrated hydrochloric acid (18 mL) at 0 ° C. The reaction solution was stirred for 15 minutes, cooled to −10 ° C., and SnCl ₂ (1.08 g, 5.696 mmol) was added slowly over 6 minutes. The reaction mixture was warmed to room temperature, stirred for a further 1.5 hours, and then poured slowly into ice (150 g). The dark brown precipitate was filtered off and washed sequentially with ethanol and diethyl ether. The aqueous filtrate was extracted with 10% CH ₃ OH / CHCl ₃ solution (50 mL × 4). The filtered precipitate was dissolved in 30% CH ₃ OH / CHCl ₃ solution (350 mL), and then the organic layer solution was passed through a short silica gel column. The organic solvent was evaporated under reduced pressure to obtain 722 mg (yield 94%) of the title compound, which was used in the next reaction without further purification.

IR (neat): 3363 and 3257 (NH ₂ ), 1755 (lactone), 1653 (pyridone), 1589, 1450, 1258, 1161 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.99 (t, J = 7.4 Hz, 3H, H-18), 1.87 (m, 2H, H-19), 3.89 (t, J = 5.6 Hz, 2H, CH ₂ Br), 4.46 (t, J = 5.6 Hz, 2H, CH ₂ CH ₂ Br), 5.28 (s, 2H, H-5), 5.42 (s, 2H, H-17), 6.47 (br s, 1H, OH), 7.29 (s, 1H, H-14), 7.45 (d, J = 9.0 Hz, 1H, H-11), 7.63 (d, J = 9.0 Hz, 1H, H-12), 8.85 ( s, 1H, H-7).

Preparation Example 6 Preparation of 9-Amino-10- (2-Bromoethoxy) -7-ethyl- (20S) -chemtothecin (Formula 5, R ¹ = CH ₂ CH ₃ )

10- (2-bromoethoxy) -7-ethyl-9-nitro- (20S) -chemtocecin (8.78 g, 16.13 mmol) in small portions over 20 minutes with stirring of concentrated hydrochloric acid (180 mL) at 0 ° C. Added. After stirring for 20 minutes and cooling the reaction solution to -10 ℃, SnCl ₂ (11.00 g, 58.07 mmol) was added slowly over 10 minutes. The reaction mixture was warmed to room temperature, stirred for a further 1.5 hours, and then poured slowly onto ice (1 kg). The yellow precipitate was filtered off and washed sequentially with ethanol and diethyl ether. The aqueous filtrate was extracted with 10% CH ₃ OH / CHCl ₃ solution (800 mL × 3). The filtered precipitate was dissolved in 10% CH ₃ OH / CHCl ₃ solution (400 mL), and then the organic layer solution was passed through a short silica gel column. The organic solvent was evaporated under reduced pressure to obtain 7.55 g (yield 91%) of the title compound, which was used in the next reaction without further purification.

IR (neat): 3460 and 3388 (NH ₂ ), 1751 (lactone), 1658 (pyridone), 1605, 1278, 1165 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.98 (t, J = 7.0 Hz, 3H, H-18), 1.34 (t, J = 7.4 Hz, 3H, CH ₂ CH ₃ ), 1.85 (m, 2H , H-19), 3.35 (q, J = 7.4 Hz, 2H, CH ₂ CH ₃ ), 3.92 (t, J = 5.2 Hz, 2H, CH ₂ Br), 4.48 (t, J = 5.2 Hz, 2H, CH ₂ CH ₂ Br), 5.28 (s, 2H, H-5), 5.41 (s, 2H, H-17), 7.23 (s, 1H, H-14), 7.52 (d, J = 9.3 Hz, 1H , H-11), 7.63 (d, J = 9.3 Hz, 1H, H-12).

Example 1 (9S) -9-ethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6, 7] Preparation of indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 1)

Method A:

9-amino-10- (2-bromoethoxy)-(20S) -chemtocecin (3.20 g, 6.58 mmol) in dry acetone (600 mL), anhydrous K ₂ CO ₃ (1.80 g, 13.02 mmol) and NaI (2.00 g, 13.34 mmol) was added and heated to reflux for 16 h under a stream of nitrogen. The reaction mixture was filtered through celite, the filtrate was washed with hot acetone and the filtrate was dried under reduced pressure. The obtained orange solid was purified by silica gel-filled MPLC (eluent: 3% CH ₃ OH / CHCl ₃ → 5% CH ₃ OH / CHCl ₃ → 10% CH ₃ OH / CHCl ₃ ) to give 2.63 g of the target compound (yield 99%). Was obtained, which was recrystallized from CH ₃ OH / CHCl ₃ solution.

mp: ＞ 300 ℃

IR (neat): 3375 (NH), 1755 (lactone), 1658 (pyridone), 1591, 1341, 1242, 1160 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.99 (t, J = 7.4 Hz, 3H, H-17), 1.87 (m, 2H, H-18), 3.48 (m, 2H, H-2), 4.24 (m, 2H, H-3), 5.28 (s, 2H, H-15), 5.42 (s, 2H, H-12), 6.48 (br s, 2H, OH, and NH), 7.27 (s, 1H , H-8), 7.34 (d, J = 9.2 Hz, 1H, H-5), 7.41 (d, J = 9.2 Hz, 1H, H-6), 8.67 (s, 1H, H-16)

FAB-MS, m / z: 406 (M + H) ⁺

Method B:

Dissolve by adding 9-amino-10- (2-bromoethoxy)-(20S) -chemtocecin (70 mg, 0.144 mmol) to DMSO (4 mL), stirring at room temperature for 20 hours, and then adding H ₂ O ( 40 ml) was added little by little. The resulting orange precipitate was filtered, dissolved in a small amount of 30% CH ₃ OH / CHCl ₃ solution, and then purified by silica gel-filled MPLC (eluent: 5% CH ₃ OH / CHCl ₃ ) to give 57 mg of the target compound (yield 98%). Got.

Example 2: (9S) -9,16-diethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4' Preparation of: 6,7] indorisino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 2)

Method A:

9-amino-10- (2-bromoethoxy) -7-ethyl- (20S) -chemtocecin (7.55 g, 14.68 mmol) in dried acetone (600 mL), anhydrous K ₂ CO ₃ (6.09 g, 44.06 mmol) and NaI (4.40 g, 29.35 mmol) were added and heated to reflux for 56 hours under a stream of nitrogen. The reaction mixture was filtered, the filtrate was evaporated to dryness under reduced pressure, and the residue was dissolved in 10% CH ₃ OH / CHCl ₃ (400 mL). The filtered yellow solid was suspended in 10% CH ₃ OH / CHCl ₃ (700 mL) and then filtered. The organic layer solution was passed through a short silica gel column. The organic solvent was evaporated under reduced pressure until a solid was formed, and the obtained concentrate was triturated using diethyl ether (200 mL) and filtered to give 6.17 g (yield 97%) of the title compound as a yellow solid. Recrystallized from CHCl ₃ solution.

mp: 249-250 ° C (dec)

IR (neat): 3318, 1756 (lactone), 1659 (pyridone), 1598, 1236, 1163 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.98 (t, J = 7.2 Hz, 3H, H-17), 1.33 (t, J = 7.5 Hz, 3H, CH ₂ CH ₃ ), 1.86 (m, 2H , H-18), 3.30-3.45 (m, 4H, CH ₂ CH ₃ and H-2), 4.20 (m, 2H, H-3), 5.27 (s, 2H, H-15), 5.34 (br s , 1H, NH), 5.42 (s, 2H, H-12), 6.46 (br s, 1H, OH), 7.22 (s, 1H, H-8), 7.32 (d, J = 9.3 Hz, 1H, H -5), 7.52 (d, J = 9.3 Hz, 1H, H-6)

FAB-MS, m / z: 434 (M + H) ⁺

Method B:

9-amino-10- (2-bromoethoxy) -7-ethyl- (20S) -chemtocecin (70 mg, 0.136 mmol) was dissolved in DMSO (4 mL), and the mixture was stirred at room temperature for 20 hours. It was added little by little to H ₂ O (40 mL). The resulting orange precipitate was filtered, dissolved in a small amount of 10% CH ₃ OH / CHCl ₃ solution, and then purified by silica gel-filled MPLC (eluent: 1% CH ₃ OH / CHCl ₃ ) to give 57 mg of the target compound (yield 97%). Got.

Example 3: (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-methyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4 ': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 3)

(9S) -9-ethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] -pyrano [3 ', 4 ': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (200 mg, 0.49 mmol) was suspended followed by methane iodide (CH ₃ I; 210 mg). , 1.48 mmol, 92 μl) was added and heated at 50 ° C. for 21 hours under a nitrogen stream. Further CH ₃ I (490 mg, 3.45 mmol, 215 μl) was added to the reaction mixture and heating was continued for 48 hours at the same temperature. The reaction mixture was cooled to room temperature and poured into hexane (100 mL). The brown liquid portion was separated, poured into water (800 mL) and extracted with CHCl ₃ (60 mL × 3). The organic layer solution was dried over anhydrous magnesium sulfate, filtered and distilled under reduced pressure. The residue was purified by silica gel-filled MPLC (eluent: 1% CH ₃ OH / CHCl ₃ ) to give 172 mg (yield 83%) of the title compound, which was recrystallized from methanol.

mp: 262 ° C (dec)

IR (neat): 3103, 1746 (lactone), 1665 (pyridone), 1608, 1573, 1505, 1363, 1240, 1116 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.99 (t, J = 7.4 Hz, 3H, H-17), 1.87 (m, 2H, H-18), 2.92 (s, 3H, NCH ₃ ), 3.22 (m, 2H, H-2), 4.31 (m, 2H, H-3), 5.23 (s, 2H, H-15), 5.41 (s, 2H, H-12), 6.48 (br s, 1H, OH), 7.25 (s, 1H, H-8), 7.37 (d, J = 9.3 Hz, 1H, H-5), 7.73 (d, J = 9.3 Hz, 1H, H-6), 8.69 (s, 1H, H-16)

FAB-MS, m / z: 420 (M + H) ⁺

Example 4: (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-ethyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4 ': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 4)

(9S) -9-ethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] -pyrano [3 ', 4 ': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (865 mg, 2.134 mmol) was suspended followed by ethane iodide (EtI; 9.98 g, 64.00 mmol, 5.12 mL) was added and heated at 85 ° C. for 48 hours under a stream of nitrogen. The reaction mixture was cooled to room temperature and the yellow precipitate was filtered and washed sequentially with DMF, diethyl ether and methanol to give 645 mg (yield 70%) of the title compound, which was recrystallized from CH ₃ OH / CHCl ₃ solution. .

mp: 257-259 ° C (dec)

IR (neat): 3216, 1749 (lactone), 1671 (pyridone), 1610, 1576, 1237, 1117 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.87 (t, J = 6.9 Hz, H-17), 1.39 (t, J = 6.9 Hz, 3H, NCH ₂ CH ₃ ), 1.86 (m, 2H, H -18), 3.01 (q, J = 6.9 Hz, 2H, NCH ₂ CH ₃ ), 3.19 (m, 2H, H-2), 4.27 (m, 2H, H-3), 5.29 (s, 2H, H -15), 5.42 (s, 2H, H-12), 6.50 (s, 1H, OH), 7.26 (s, 1H, H-8), 7.39 (d, J = 9.3 Hz, 1H, H-5) , 7.73 (d, J = 9.3 Hz, 1H, H-6), 8.55 (s, 1H, H-16)

FAB-MS, m / z: 434 (M + H) ⁺

Example 5: (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-cyanomethyl-12H-1,4-oxazino [3,2-f] pyrano [3 ' , 4 ': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 5)

(9S) -9-ethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] -pyrano [3 ', in dried DMF (38 mL). 4 ': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (1.62 g, 3.996 mmol) was suspended followed by bromoacetonitrile (15.15 g, 126 mmol, 8.8 mL) was added and heated at 80 ° C. for 20 hours under a stream of nitrogen. The reaction mixture was vacuum distilled and the residue was purified by silica gel-filled MPLC (eluent: 10% CH ₃ OH / CHCl _3- > 20% CH ₃ OH / CHCl ₃ ) and recrystallized from 20% CH ₃ OH / CHCl ₃ solution. 1.74 g (98%) of target compounds were obtained.

mp: 248 ~ 249 ℃ (dec)

IR (neat): 3318, 1749 (lactone), 1661 (pyridone), 1600, 1560, 1236, 1157 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.87 (t, J = 7.3 Hz, 3H, H-17), 1.86 (m, 2H, H-18), 3.43 (m, 2H, H-2), 4.41 (s, 2H, CH ₂ CN), 4.46 (m, 2H, H-3), 5.24 (s, 2H, H-15), 5.41 (s, 2H, H-12), 6.51 (br s, 1H , OH), 7.27 (s, 1H, H-8), 7.45 (d, J = 9.3 Hz, 1H, H-5), 7.86 (d, J = 9.3 Hz, 1H, H-6), 8.59 (s , 1H, H-16)

FAB-MS, m / z: 445 (M + H) ⁺

Example 6: (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1- (2-aminoethyl) -12H-1,4-oxazino [3,2-f] pyrano Preparation of [3 ', 4': 6,7] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione acetate salt (Compound No. 6)

To glacial acetic acid (180 mL) (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-cyanomethyl-12H-1,4-oxazino [3,2-f] -pyrano [3 ', 4': 6,7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (1.00 g, 2.250 mmol) was added to dissolve and then Nickel (50% slurry in H ₂ O, 32 ml) was added and purified three times with hydrogen gas (H ₂ ). A hydrogen balloon was mounted and stirred at room temperature under hydrogen pressure for 12 hours, and then filtered through celite. The filtrate was concentrated under reduced pressure, H ₂ O (250 mL) was added to a red residue, and the mixture was filtered through celite. The celite filter cake was extracted with 20% CH ₃ OH / CHCl ₃ solution (100 mL) to give (9S) -9-ethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [ 3,2-f] pyrano [3 ', 4': 6,7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 1; 580 mg, yield 64%). The filtrate was concentrated under reduced pressure and purified by C ₁₈ reversed phase MPLC (eluent: 20% CH ₃ OH / H ₂ O with 1% acetic acid → 50% CH ₃ OH / H ₂ O with 1% acetic acid) to obtain the title compound (350 mg) as a yellow solid. (Yield 31%) was obtained.

mp: 238 ° C (dec, dark color from 138 ° C)

IR (neat): 3101, 1747 (lactone), 1659 (pyridone), 1597, 1572, 1415, 1243, 1051 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.87 (t, J = 7.1 Hz, 3H, H-17), 1.78 (s, 3H, CH ₃ CO ₂ H), 1.84 (m, 2H, H-18 ), 3.04 (m, 2H, CH ₂ NH ₂ ), 3.20 (m, 2H, CH ₂ CH ₂ NH ₂ ), 3.40 (m, 2H, H-2), 4.26 (m, 2H, H-3), 5.27 (s, 2H, H-15), 5.41 (s, 2H, H-12), 6.50 (br s, 1H, OH), 7.26 (s, 1H, H-8), 7.37 (d, J = 9.3 Hz, 1H, H-5), 7.72 (d, J = 9.3 Hz, 1H, H-6), 8.87 (s, 1H, H-16)

FAB-MS, m / z: 449 (M + H-CH ₃ CO ₂ H) ⁺

Example 7: (9S) -9,16-diethyl-2,3-dihydro-9-hydroxy-1-methyl-12H-1,4-oxazino [3,2-f] pyrano [3 Preparation of ', 4': 6,7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (Compound No. 7)

(9S) -9,16-diethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] -pyrano [3] in dry DMF (100 mL). ', 4': 6,7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione (3.03 g, 6.99 mmol) was added to dissolve and then methane iodide ( 29.80 g, 210 mmol, 9.2 mL) was added and heated at 120 ° C. for 18 hours under a nitrogen stream. The reaction mixture was cooled to room temperature, vacuum distilled, purified by silica gel-filled MPLC (eluent: 1% CH ₃ OH / CHCl ₃ → 2% CH ₃ OH / CHCl ₃ → 3% CH ₃ OH / CHCl ₃ ), and yellow. 2.037 g (65% yield) of the desired compound in the solid phase were obtained, which was recrystallized from an EtOH / CHCl ₃ solution.

mp: 250-251 ° C (dec)

IR (neat): 3286, 1745 (lactone), 1657 (pyridone), 1598, 1564, 1232, 1157 cm ^-1

¹ H NMR (DMSO-d ₆ ): δ 0.87 (t, J = 7.2 Hz, 1.5H, 0.5H-17), 0.88 (t, J = 7.2 Hz, 1.5H, 0.5H-17), 1.10 ( t, J = 6.9 Hz, 1.5H, 0.5CH ₂ CH ₃ ), 1.11 (t, J = 6.9 Hz, 1.5H, 0.5CH ₂ CH ₃ ), 1.86 (m, 2H, H-18), 2.68 (s , 3H, NCH ₃ ), 3.01 (m, 1H, H-2), 3.20 to 3.42 (m, 2H, CH ₂ CH ₃ ), 3.96 (m, 1H, H-2), 4.35 (m, 2H, H -3), 5.24 (d, J _{a, b} = 18.6 Hz, 1H, H-15), 5.37 (d, J _{a, b} = 18.6 Hz, 1H, H-15), 5.43 (s, 2H, H- 12), 6.47 (br s, 1H, OH), 7.24 (s, 1H, H-8), 7.38 (d, J = 9.3 Hz, 1H, H-5), 7.78 (d, J = 9.3 Hz, 1H , H-6)

FAB-MS, m / z: 448 (M + H) ⁺

The following formulation examples illustrate the general pharmaceutical composition according to the present invention, in which the active ingredient in each case represents the hexacyclic camptothecin derivative represented by Formula 1, and in some cases other compounds included in the present invention. It may be replaced by an effective dose equivalent to.

Formulation Example 1 Tablet

250 g of the active ingredient was mixed with 175.9 g of lactose, 180 g of potato starch, and 32 g of colloidal silicic acid. Then, 10% gelatin solution was added to the mixture, which was ground and passed through 14 mesh. Then, after drying, the mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate to each was made into a tablet.

Formulation Example 2: Injection Solution

1 g of active ingredient and 0.1 g of lactic acid were dissolved in distilled water to obtain a 100 ml solution. 1 ml of this solution contained 10 mg of the active ingredient, which was placed in an ampoule and sterilized at 20 ° C. for 30 minutes.

As described above, the hexacyclic camptothecin derivative represented by Formula 1 according to the present invention and a pharmaceutically acceptable salt thereof exhibited an effective inhibitory effect on topoisomerase I. For the biological activity effect was measured by the following method.

Analysis of Cleavage Complex Formation to Measure Inhibitory Activity of Phase Isomerase I

Hsiang et al., J. Biol. The same procedure as described in Chem., 1985, 260, 14873 was carried out with slight modifications. In this assay, all reaction buffers contained 10 mM Tris-HCl (pH 7.5), 1 mM EDTA, 150 mM NaCl, 0.1% BSA, 0.1 mM spermidine and 5% glycerol. In addition, each reaction mixture (20 µl) contained 250 ng of supercoiled DNA pHOT1, a phase isomerase I 1 enzyme unit and the compound to be tested. The reaction mixture was reacted at 37 ° C. for 30 minutes, followed by addition of a reaction termination buffer solution / loading dye reagent (including SDS 2.5%, Picol 15%, bromophenol blue 0.25% and xylene cyanole 0.25%). Terminated. Then, 5 μl of proteinase K (0.75 mg / ml) was added and reacted at 37 ° C. for 60 minutes. Each reaction mixture was then electrophoresed for 6 hours using a developing solution comprising 40 mM Tris-acetate and 1 mM EDTA at 40 volts on 1% agarose gel. On the other hand, the gel and the developing solution contains 0.5 μg / ml ethidium bromide. After the electrophoresis, the gel was photographed using ultraviolet light and the ratio of II DNA to total DNA was quantified using a BIO-PROFIL phase analysis system. IC ₅₀ in Table 1 shows the compound concentration when II DNA is formed by 50% by phase isomerase I, which is a quantal probit analysis according to pharmaceutical calculation using a computer program. Calculated by

Test compound IC ₅₀ (μM) Compound number 2 4.7 Compound number 4 11.8 Camptothecin 40.2 Topotecan 20.2

In Vitro Cytotoxicity Test

Five types of human cancer cell lines were performed by MTT analysis. The cell lines were cultured in a 37 ° C. incubator with 5% carbon dioxide using RPMI-1640 culture medium, incubated with 10% fetal bovine serum, penicillin (100 units / ml) and streptomycin (100 μg / ml) inactivated by heat treatment. ) Is included. Single cell suspensions were then obtained using trypsin treatment and pipette dissociation methods. The suspension was then transferred to 96-well microtiter plates and subjected to MTT analysis to determine the growth curves to determine the number of each cell line cell. The test compound solution in DMSO was diluted 10 fold in fresh culture. Cells were seeded into each plate well with 180 μl of culture, and then 8 different concentrations of solution of the compound to be tested were added to each well above. The plates were then reacted for 4 days at 37 ° C. using a thermostat with 5% carbon dioxide, then 0.1 mg of MTT was added to each well and subsequently for 4 hours in the thermostat. The plate was then centrifuged at 1000 rpm for 10 minutes, supernatant was taken, and then 150 μl of DMSO was added to the wells to dissolve formazan crystals, and Elisa reader (Dynatech, MR 5000). The plate was analyzed directly at 550 nm using. IC ₅₀ in Table 2 below shows compound concentrations that reduce 50% of viable cells, which are calculated by quantal probit analysis according to pharmaceutical calculations using a computer program.

Test compound IC ₅₀ (nM) WiDr MKN45 A549 SKOV3 H128 Compound number 2 15.9 19.7 14.1 14.0 7.1 Compound number 4 21.2 18.9 17.0 18.7 12.9 Camptothecin 17.0 16.9 13.8 20.2 18.3 Topotecan 56.2 38.3 53.5 48.1 31.3

As described above, the hexacyclic camptothecin derivative represented by Formula 1 according to the present invention and its pharmaceutically acceptable salts have high anticancer activity against human cancer cell lines and inhibitory activity against phase isomerase I. It is excellent and effective as an anticancer agent.

Claims (6)

A hexacyclic camptothecin derivative or a pharmaceutically acceptable salt thereof, characterized in that represented by the following formula (1). Formula 1 In Formula 1 above: R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or a C ₃ -C ₆ cycloalkyl group; R ² denotes a cyano group or an aminoalkyl group of C ₁ ~C ₇ a represents a hydrogen _{atom, C 1 ~C 6, C 1} ~C 6. According to claim 1, wherein the compound represented by the formula (1) (9S) -9-ethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6,7] indoridge No [1,2-b] quinoline-10,13 (9H, 15H) -dione, (9S) -9,16-diethyl-2,3-dihydro-9-hydroxy-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6,7 ] Indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione, (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-methyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6, 7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione, (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-ethyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6, 7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione, (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1-cyanomethyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4': 6,7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione, (9S) -9-ethyl-2,3-dihydro-9-hydroxy-1- (2-aminoethyl) -12H-1,4-oxazino [3,2-f] pyrano [3 ', 4 ': 6,7] indorizino [1,2-b] quinoline-10,13 (9H, 15H) -dione acetate salt, and (9S) -9,16-diethyl-2,3-dihydro-9-hydroxy-1-methyl-12H-1,4-oxazino [3,2-f] pyrano [3 ', 4' A hexacyclic camptothecin derivative, characterized in that it is selected from: 6,7] indorigino [1,2-b] quinoline-10,13 (9H, 15H) -dione. A pharmaceutical composition effective for inhibiting topoisomerase I, characterized in that it contains a hexacyclic camptothecin derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof. Formula 1 In Formula 1 above: R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or a C ₃ -C ₆ cycloalkyl group; R ² represents a cyano group or an aminoalkyl group of C ₁ ~C ₇ a represents a hydrogen _{atom, C 1 ~C 6, C 1} ~C 6. An anticancer agent characterized in that it contains a hexacyclic camptothecin derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof. Formula 1 In Formula 1 above: R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or a C ₃ -C ₆ cycloalkyl group; R ² represents a cyano group or an aminoalkyl group of C ₁ ~C ₇ a represents a hydrogen _{atom, C 1 ~C 6, C 1} ~C 6. i) performing a bromination reaction with a compound represented by the following formula (2) in the presence of a base to prepare a brominated compound represented by the following formula (3); Ii) nitriding the brominated compound represented by Formula 3 in the presence of nitric acid or nitric acid and sulfuric acid to prepare a nitro compound represented by Formula 4 below; Iii) preparing a amine compound represented by the following Chemical Formula 5 by performing a reduction reaction on the nitro compound represented by Chemical Formula 4 in the presence of a metal or metal salt for reduction reaction; Iii) a process for preparing the hexacyclic camptothecin derivative represented by the following formula (1) from the amine compound represented by the formula (5). Formula 1 In the above formula: R ¹ represents a hydrogen atom, a C ₁ -C ₆ alkyl group or a C ₃ -C ₆ cycloalkyl group; R ² represents a cyano group or an aminoalkyl group of C ₁ ~C ₇ a represents a hydrogen _{atom, C 1 ~C 6, C 1} ~C 6. The process of claim 5, wherein Preparing a compound represented by the formula (1) wherein R ² is a hydrogen atom from the amino compound represented by the formula (5), Reacting an alkyl halide or cyanoalkyl halide with a compound represented by formula (1) wherein R ² is a hydrogen atom to prepare a compound represented by formula (1) wherein R ² is an alkyl group or cyanoalkyl group, And a process for preparing the compound represented by formula (1) wherein R ² is an aminoalkyl group by reduction of the compound represented by formula (1) wherein R ² is cyanoalkyl group with molecular hydrogen (H ₂ ) under an organic or inorganic acid catalyst. The manufacturing method to make.