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WO1999032499A1 - 4'-O-DEMETHYL-EPIPODOPHYLLOTOXIN-β-D-GLUCOSIDE ACETAL DERIVATIVES - Google Patents

4'-O-DEMETHYL-EPIPODOPHYLLOTOXIN-β-D-GLUCOSIDE ACETAL DERIVATIVES Download PDF

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Publication number
WO1999032499A1
WO1999032499A1 PCT/KR1998/000452 KR9800452W WO9932499A1 WO 1999032499 A1 WO1999032499 A1 WO 1999032499A1 KR 9800452 W KR9800452 W KR 9800452W WO 9932499 A1 WO9932499 A1 WO 9932499A1
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Prior art keywords
glucoside
epipodophyllotoxin
demethyl
phenyl
compound
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PCT/KR1998/000452
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French (fr)
Inventor
Zae Sung No
Bok Ju Song
Eun Yi Song
Jong Woong Ahn
Chong Ock Lee
Sang Un Choi
Kyu Lim
Byung Doo Hwang
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Korea Research Institute Of Chemical Technology
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Publication of WO1999032499A1 publication Critical patent/WO1999032499A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms

Definitions

  • the present invention relates to novel 4 '-O-demethyl- epipodophyllotoxin-3-D-glucoside acetal derivatives having an excellent anticancer activity, a process for the preparation thereof and an anticancer composition containing same as an active ingredient.
  • etoposide and teniposide have severe cytotoxicities and are insoluble in water, and, therefore, various attempts have been made to develop a novel anticancer agent by modifying etoposide.
  • etoposide and teniposide have severe cytotoxicities and are insoluble in water, and, therefore, various attempts have been made to develop a novel anticancer agent by modifying etoposide.
  • Bristol-Myers Squibb Co. in the U.S. and Microbial Chemistry Research Foundation in Japan have developed ETOPOPHOSTM and NK-611, respectively.
  • ETOPOPHOSTM was approved as an anticancer drug in 1997 and NK-611 is now said to be in a clinical test stage ( see GB Patent No. 2,207,674 A and EP Patent No. 0,196,618).
  • the present inventors have endeavored to develop compounds having a high potency against cancer cells and improved pharmacokinetic properties; and have unexpectedly found that 4 '-0-demethyl-epipodophyllotoxin-/3-D-glucoside derivatives substituted with a hydrophobic group containing a double or triple bond, or an alkoxyalkyl group in the glucosidic 4,6-diol positions exhibit superior anticancer activities .
  • R 1 and R 2 are, each independently, hydrogen, a C 2 _ 12 alkenyl, C 2 _ 12 alkynyl, C 2 _ 12 alkoxylalkyl or cyclopropyl group optionally substituted with one or more radicals selected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups , with the proviso that R 1 and R 2 are not hydrogen simultaneously; or
  • R 1 and R 2 together with the carbon atom to which they are attached, form an unsaturated C 5 _ 15 ring optionally substituted with one or more radicals selected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups.
  • the compound of formula(I) may be prepared by a process which comprises reacting 4 ' -O-demethy 1- epipodophyllotoxin-/3-D-glucoside of formula (II) with an aldehyde of formula (III) or an acetal of formula (IV) in a suitable solvent in the presence of a catalyst as shown in Scheme 1.
  • R 1 and R 2 have the same meanings as defined above, and R 3 is a C,_ 3 alkyl group.
  • a compound of formula(II) may be reacted with a compound of formula (III) or (IV) in a molar ratio ranging from 1:5 to 1:20, preferably from 1:10 to 1:15, at a temperature ranging from 0 to 80°C, preferably at room temperature, for 1 to 48 hours, preferably from 3 to 24 hours.
  • Exemplary solvents which may be suitably used in the process of the present invention are nitromethane, dichloro ethane , chloroform, diethyl ether, tetrahydrofuran, dimethoxyethane , acetonitrile and dimethylformamide, wherein anhydrous acetonitrile and anhydrous nitromethane are preferred.
  • the catalyst which may be used in practicing the present invention is an acid such as p-toluenesulfonic acid, methanesulfonic acid, zinc chloride and an acidic resin, wherein p-toluenesulfonic acid and zinc chloride are preferred.
  • the catalyst may be used in an amount ranging from 3 to 30 wt%, preferably from 5 to 10 wt% based on the weight of the compound of formula(II).
  • 3A may be added to the reaction system to remove water produced during the reaction.
  • 4A molecular sieve is used in an amount of 200 wt% based on the weight of the compound of formula(H).
  • reaction may be terminated by adding an organic base such as trialkylamine or a pyridine derivative in an amount slightly excess of the acid catalyst employed.
  • 4'-0-demethyl-epipodophyllotoxin-/3-D-glucoside of formula(II) may be prepared in accordance with a known method(see P. Allevi, M. Anastasia, P. Ciuffreda, A. M. Sanvito and P. Macdonald, Tet . Lett . , 1992, 33, 4831-4834; S. Hashimoto, T. Hyundai, and S. Ikegami, Tet. Lett.. 1991, 32, 1653-1654).
  • aldehydes of formula(III) which may be suitably used in the process of the present invention include cyclopropanecarboxaldehyde, phenylpropagylaldehyde, methacrolein, ⁇ -methyl-trans-cinnamaldehyde, 1,2,3,6- tetrahydrobenzaldehyde, 3-methyl-2-butenaldehyde and the like.
  • acetal compound of formula (IV) are acroleindimethylacetal, 3-butenal-diethylacetal, 3- methoxybutyraldehyde dimethylacetal, 2-butyl-l-aldehyde diethylacetal, 3-ethoxypropionaldehyde diethylacetal and 1,1, 3-trimethoxypropane .
  • the present invention also includes within its scope pharmaceutical compositions comprising one or more of the compounds of formula(I) as an active ingredient, in association with pharmaceutically acceptable carriers, excipients or other additives, if necessary.
  • compositions of the present invention may be formulated for administration orally or, preferably, by injection.
  • the composition for oral administration may take various forms such as tablets and gelatin capsules, which may contain conventional additives such as a diluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), a lubricant (e.g., silica, talc, stearic acid or its magnesium and calcium salts and polyethylene glycol).
  • a diluent e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine
  • a lubricant e.g., silica, talc, stearic acid or its magnesium and calcium salts and polyethylene glycol.
  • the composition may further comprise a binder (e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose and polyvinyl picolidine) and optionally a disintegrant (e.g., starch, agar and alginic acid or its sodium salt), absorbent, colorant, flavor and sweetener.
  • a binder e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose and polyvinyl picolidine
  • a disintegrant e.g., starch, agar and alginic acid or its sodium salt
  • the amount of the active ingredient actually administered should be determined in light of various relevant factors including the condition to be treated, the chosen route of administration, the age and weight of the individual patient, and the severity of the patient's symptoms; and, therefore, the dosage suggested above should not be construed to limit the scope of the invention in any way.
  • Example 1 Synthesis of 4 ' -O-demethyl-epipodophyllotoxin- /3-D- ( alkenylidene or alkynylidene ) -glucoside derivatives ( Compounds 1-8 ) 200 mg(0.36 mmol) of 4 ' -O-demethyl-epipodophyllotoxin- 3-D-glucoside was dissolved in 20 ml of anhydrous acetonitrile in the presence of 400 mg of 4A molecular sieve and added thereto were 20 mg of p-toluenesulfonic acid and an excess amount ( 2.0-3.5 mmol) of acrolein dimethylacetal (Compound 1), 3-butenaldiethylacetal (Compound 2), 2-butyne-1-aldehyde diethylacetal (Compound 3), phenylpropagylaldehyde( Compound 4), ⁇ -methyl-trans- cinnamaldehyde ( Compound 1
  • Compound 2 4 '-0-demethyl-epipodophyllotoxin-/3-D-(3- butenylidene) -glucoside;
  • Compound 8 4 '-0-demethyl-epipodophyllotoxin-/3-D-(2- methyl-2-propenylidene) -glucoside;
  • Test Example 1 in vitro anticancer activity
  • Step 1 cell cultivation
  • Cancer cell lines derived from human, i.e., A-549 (non- molding type cell lung cancer), SK-OV-3 (adenocarcinoma, ovary malignant hydroperitoneum) , SK-MEL-2 (malignant melanoma, transferred to femoral skin), XF498(cancer of central nervous system) and HCT15 (colonic cancer) were obtained from U.S. National Cancer Institute and subcultured in Korea Research Institute of Chemical Technology. These cell lines were cultured in a RPMI (Rosewell Park Medium) 1640 culture medium enriched with 5% fetal bovine serum at a constant temperature of 37°C and constant humidity of 98% under an atmosphere of 5% C0 2 .
  • RPMI Rosewell Park Medium
  • Step 2 in vitro anticancer activity
  • the in vitro anticancer activity of the compound of the present invention was assessed in accordance with SRB (sulforhodamin B) assay method developed by U.S. National Cancer Institute in 1989.
  • the subcultured cells were treated with a tripsin-CDTA ⁇ olution to separate the cells, which were added to the wells of a 96 well microplate(Falcon Co., U.S.) in an amount of 5 x 10 3 (A549, HCT15), 1 x 10 4 (SK-MEL-2 , XF498), 2 x 10 4 (SK-OV-3) cells per each wells.
  • the cells were incubated under a C0 2 atmosphere for 24 hours to be affixed on the well bottom and the culture medium was removed with an aspirator.
  • the Compounds 1 to 12 of the present invention and etoposide(BMS, U.S.A) were used as test compounds. Each of the test compounds was dissolved in water to obtain a test solution. 0.5% dimethylsulfoxide was used to dissolve the compounds, if necessary. The test solution was diluted with the culture medium(pH 7.2), e.g., by a factor of 1, 10 " ⁇ 10" 2 , 10" 3 , 10" A and 10 -5 . After filtering through a 0.22ml filter, 100 ml(x3) of each of the diluted test solutions was added to the wells and the mixture was cultured for another 48 hours.
  • TCA trichloroacetic acid
  • the anticancer activities (A. . ) of the compounds were calculated by equation (1) or (2).
  • A. A. [ (T-Tz)/(C-Tz) ] X 100 (Tz>T) Eq. (1)
  • A. A. [(T-Tz)/Tz] x 100 (Tz ⁇ T) Eq. (2)
  • Tz is the number of cells before adding a test compound without incubation
  • C is the number of cells after incubation for 48 hours in the absence of a test compound
  • T is the number of cells after incubation for 48 hours the cell in the presence of a test compound.
  • ED 50 i.e., the concentration of a compound to inhibit the growth of a cancer cell by 50% was calculated by regression method using LOTUS program.
  • Test Example 2 Inhibition of human topoisomera ⁇ e II
  • Step 1 Preparation of topoisomerase II HeLa cell line provided by ATCC(American Type Culture Collection) was used in preparing topoiso erase II of the cancer cell.
  • the HeLa cells were cultured in a RPMI 1640 culture medium enriched with 10% fetal bovine serum at 37°C under an atmosphere of 5% C0 2 and the cultured HeLa cells were collected. Added thereto was a 3-fold volume of 50 mM KH 2 PO A buffer solution(pH 7.0) containing 1 mM ethylenediamine tetraacetic acid(EDTA), ImM mercaptoethanol, 0.5 mM phenylmethylsulfonylchloride(PMSF) and 10% glycerol and the resulting mixture was homogenized with Polytron* for 30 seconds in ice bath.
  • EDTA ethylenediamine tetraacetic acid
  • PMSF phenylmethylsulfonylchloride
  • a 2M KH 2 PO buffer solution(pH 7.0) was added to a KH 2 PO concentration of 0.3M and the resultant was homogenized for 30 seconds, placed in an ice bath for 1 hour, and then, centrifuged at 30,000 xg for 30 minutes. The supernatant was purified by phosphocellulose column chromatography(eluent : linear gradient from 0.3M to 0.5M KH 2 P0 4 buffer(pH 7.0) to obtain topoisomerase II.
  • Step 2 P unknotting assay
  • IC 50 values of the test compounds for topoisomerase II were measured by an unknotting assay.
  • Topo II obtained above and 0.4 ⁇ g of P knotted DNA were added to a 50 mM N-[ 2-hydroxyethyl]-piperazine-N'-[ 2- ethanesulfonic acid] (HEPES) buffer solution (pH 7.0) containing 50 mM KC1, 0.1 mM EDTA, 100 mM NaCl, 10 mM MgCl 2 , 100 ⁇ g/ml of fetal bovine serum albumin and 1 mM adenosine triphosphate(ATP) to a final volume of 20 ⁇ l and the mixture wa ⁇ reacted at 37 °C for 30 minutes.
  • HEPES 2-hydroxyethyl]-piperazine-N'-[ 2- ethanesulfonic acid]
  • the reaction was terminated by adding sodium dodecyl sulfate(SDS) to a SDS concentration of 1%.
  • SDS sodium dodecyl sulfate
  • the resultant was subjected to electrophoresis using 1% agarose gel equilibrated with a 50 mM tris borate buffer solution(pH 8.3) containing 2.5mM EDTA, and then, dyed with ethidium bromide .
  • Topo II The activity of Topo II was assessed using photographs taken under UV radiation. The Topo II activity measured when 50% of 0.4 ⁇ g of knotted P A DNA was unknotted was defined as 1 unit activity. IC 50 was the concentration of a compound to inhibit the activity of Topo II by 50%. The results are shown in Table II.
  • Test Example 3 In. vivo anticancer activity
  • a dose of 1 x 10 s cells/0. lml of leukemia L1210 cell subcultured in DBA/2 mouse was injected into the abdominal cavity of each of the mice.
  • Two test compound ⁇ , etoposide and Compound 1 of the present invention were injected into the abdominal cavities of mice of respective test group in the doses shown in Table III, successively at 1st, 2nd, 3rd, 4th and 5th days from the leukemia cell injection.
  • mice were observed daily and the survival period of each mouse was checked.
  • the anticancer activity was assessed by calculating the ratio (I.R.) by equation 3.
  • I.R. 100 x (A t - A c )/A c Eq (3) wherein, A t is the average survival days of mice in a test group and A c is the average survival days of mice in the contro1 group.
  • the novel 4 '-O-demethy1- epipodophyllotoxin-/3-D-glucoside derivative of formula(I) of the present invention possesses an excellent anticancer activity against various cancer cells, much higher than that of etoposide.
  • the inventive compounds can therefore be used as a drug for treating cancers.

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Abstract

A 4'-O-demethyl-epipodophyllotoxin-β-D-glucoside acetal derivative of formula (I), wherein R?1 and R2¿ are, each independently, hydrogen, a C¿2-12? alkenyl, a C2-12 alkynyl, C2-12 alkoxylalkyl or cyclopropyl group optionally substituted with one or more radicals selected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups, with the proviso that R?1 and R2¿ are not hydrogen simultaneously; or R?1 and R2¿, together with the carbon atom to which they are attached, form an unsaturated C¿5-15? ring optionally substituted with one or more radicals selected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups.

Description

' -O-DEMETHYL-EPIP0D0PHYI-I-OTOXIN-5-D-GI.UC0SIDE ACETA DERIVATIVES
Field of the Invention
The present invention relates to novel 4 '-O-demethyl- epipodophyllotoxin-3-D-glucoside acetal derivatives having an excellent anticancer activity, a process for the preparation thereof and an anticancer composition containing same as an active ingredient.
Description of the Prior Art
Etoposide and teniposide, semi-synthetic anti-tumor drugs from a natural product, podophyllotoxin isolated from mandrake(PodophyHum peltatum L.) exhibit good anticancer activities when employed in radiotherapy of various cancers such as lung cancer, malignant lymphoma, leukemia and testis cancer ( see C. Keller-Julsen, M. Kuhn, A. von Wartburg, J. Med. Chem. , 1971, 14(10), 936-940 and U.S. Patent No. 3,524,844). These compounds are known to prevent DNA synthesis by inhibiting human DNA topoisomerase II, thereby suppressing the growth of cancer cells (see J. C. Wang, J. Biol. Chem., 1991, 266(11), 6659-62).
However, etoposide and teniposide have severe cytotoxicities and are insoluble in water, and, therefore, various attempts have been made to develop a novel anticancer agent by modifying etoposide. For example, Bristol-Myers Squibb Co. in the U.S. and Microbial Chemistry Research Foundation in Japan have developed ETOPOPHOS™ and NK-611, respectively. ETOPOPHOS™ was approved as an anticancer drug in 1997 and NK-611 is now said to be in a clinical test stage ( see GB Patent No. 2,207,674 A and EP Patent No. 0,196,618).
It has been known that modification of the 4,6-acetal functional group of the glucoside part of etoposide directly leads to an altered anticancer activity. However, only a limited number of etoposide derivatives having unsaturated and alkoxy acetal functional groups, e.g., those derived from crotonaldehyde and cinnamaldehyde, have been reported in the literature ( see U.S. Patent No. 3,524,844 and C. Keller-Julsen, M. Kuhn, A. von Wartburg, J. Med. Chem. , 1971, 14(10), 936-940).
The present inventors have endeavored to develop compounds having a high potency against cancer cells and improved pharmacokinetic properties; and have unexpectedly found that 4 '-0-demethyl-epipodophyllotoxin-/3-D-glucoside derivatives substituted with a hydrophobic group containing a double or triple bond, or an alkoxyalkyl group in the glucosidic 4,6-diol positions exhibit superior anticancer activities .
Summary of the Invention
It is, therefore, a primary object of the present invention to provide novel 4 '-O-demethyl-epipodophyllo- toxin---D-glucoside acetal derivatives having an excellent anticancer activities.
It is another object of the present invention to provide an anticancer composition containing one or more of the inventive compounds as an active ingredient.
It is a further object of the present invention to provide a process for the preparation of the inventive novel compounds .
In accordance with the present invention, there is provided a 4 '-O-demethyl-epipodophyllotoxin- -D-glucoside acetal derivative of formula (I):
Figure imgf000004_0001
wherein
R1 and R2 are, each independently, hydrogen, a C2_12 alkenyl, C2_12 alkynyl, C2_12 alkoxylalkyl or cyclopropyl group optionally substituted with one or more radicals selected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups , with the proviso that R1 and R2 are not hydrogen simultaneously; or
R1 and R2, together with the carbon atom to which they are attached, form an unsaturated C5_15 ring optionally substituted with one or more radicals selected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups.
Detailed Description of the Invention
Among the compounds of the present invention, preferred are:
4 ' -O-demethyl-epipodophyllotoxin-β-D- ( 2 - propenylidene)-glucoside;
4 '-0-demethyl-epipodophyllotoxin-jS-D-( 3-butenylidene)- glucoside;
4 ' -O-demethy 1 -epipodophy llotoxin-β-D- ( 2 -butyny lidene ) - glucoside; 4 ' -O-demethy 1-epipodophyllotoxin- -D- ( 3-phenyl-2- propyny lidene ) -glucoside ;
4 ' -0-demethyl-epipodophyllotoxin-3-D- ( 2-methyl-3- phenyl-2-propeny lidene) -glucoside;
4 ' -O-demethyl-epipodophyllotoxin-β-D- ( 3 - cyclohexenylidene) -glucoside;
4 ' -0-demethyl-epipodophyllotoxin-3-D- ( 3-methyl-2- butenylidene) -glucoside;
4 ' -O-demethyl-epipodophyllotoxin-β-D- ( 2-methyl-2- propenylidene ) -glucoside ; 4 ' -O-demethy 1 -epipodophy Ho toxin - -D-( 3-methoxy-l- butylidene ) -glucoside;
4 ' -0-demethyl-epipodophyllotoxin-3-D- ( 3-ethoxy-l- propylidene ) -glucos ide ; 4 ' -O-demethyl-epipodophyllotoxin-β-D- ( 3-methoxy-l- propy lidene) -glucoside; and
4 ' -0-demethyl-epipodophyllotoxin-/3-D- ( cyclopropylmethylidene ) -glucoside .
The compound of formula(I) may be prepared by a process which comprises reacting 4 ' -O-demethy 1- epipodophyllotoxin-/3-D-glucoside of formula (II) with an aldehyde of formula (III) or an acetal of formula (IV) in a suitable solvent in the presence of a catalyst as shown in Scheme 1.
Scheme 1
Figure imgf000006_0001
(D) (IV) (I)
wherein, R1 and R2 have the same meanings as defined above, and R3 is a C,_3 alkyl group.
In the above reaction, a compound of formula(II) may be reacted with a compound of formula (III) or (IV) in a molar ratio ranging from 1:5 to 1:20, preferably from 1:10 to 1:15, at a temperature ranging from 0 to 80°C, preferably at room temperature, for 1 to 48 hours, preferably from 3 to 24 hours.
Exemplary solvents which may be suitably used in the process of the present invention are nitromethane, dichloro ethane , chloroform, diethyl ether, tetrahydrofuran, dimethoxyethane , acetonitrile and dimethylformamide, wherein anhydrous acetonitrile and anhydrous nitromethane are preferred. The catalyst which may be used in practicing the present invention is an acid such as p-toluenesulfonic acid, methanesulfonic acid, zinc chloride and an acidic resin, wherein p-toluenesulfonic acid and zinc chloride are preferred. The catalyst may be used in an amount ranging from 3 to 30 wt%, preferably from 5 to 10 wt% based on the weight of the compound of formula(II).
Further, an excess amount of molecular sieves (4A or
3A) may be added to the reaction system to remove water produced during the reaction. Preferably, 4A molecular sieve is used in an amount of 200 wt% based on the weight of the compound of formula(H).
The reaction may be terminated by adding an organic base such as trialkylamine or a pyridine derivative in an amount slightly excess of the acid catalyst employed. 4'-0-demethyl-epipodophyllotoxin-/3-D-glucoside of formula(II) may be prepared in accordance with a known method(see P. Allevi, M. Anastasia, P. Ciuffreda, A. M. Sanvito and P. Macdonald, Tet . Lett . , 1992, 33, 4831-4834; S. Hashimoto, T. Honda, and S. Ikegami, Tet. Lett.. 1991, 32, 1653-1654).
Exemplary aldehydes of formula(III) which may be suitably used in the process of the present invention include cyclopropanecarboxaldehyde, phenylpropagylaldehyde, methacrolein, α-methyl-trans-cinnamaldehyde, 1,2,3,6- tetrahydrobenzaldehyde, 3-methyl-2-butenaldehyde and the like. Representative acetal compound of formula (IV) are acroleindimethylacetal, 3-butenal-diethylacetal, 3- methoxybutyraldehyde dimethylacetal, 2-butyl-l-aldehyde diethylacetal, 3-ethoxypropionaldehyde diethylacetal and 1,1, 3-trimethoxypropane .
The present invention also includes within its scope pharmaceutical compositions comprising one or more of the compounds of formula(I) as an active ingredient, in association with pharmaceutically acceptable carriers, excipients or other additives, if necessary.
The pharmaceutical compositions of the present invention may be formulated for administration orally or, preferably, by injection. The composition for oral administration may take various forms such as tablets and gelatin capsules, which may contain conventional additives such as a diluent (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine), a lubricant (e.g., silica, talc, stearic acid or its magnesium and calcium salts and polyethylene glycol). In the tablet form, the composition may further comprise a binder (e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methyl cellulose, sodium carboxymethyl cellulose and polyvinyl picolidine) and optionally a disintegrant (e.g., starch, agar and alginic acid or its sodium salt), absorbent, colorant, flavor and sweetener. The composition for injection may be an isotonic solution or a suspension. The inventive pharmaceutical composition may be administered daily. A typical daily dose of the active ingredient ranges from about 1 to 100 g/kg, preferably 1 to 20 mg/kg, and can be administered in a single dose or in divided doses. However, it should be understood that the amount of the active ingredient actually administered should be determined in light of various relevant factors including the condition to be treated, the chosen route of administration, the age and weight of the individual patient, and the severity of the patient's symptoms; and, therefore, the dosage suggested above should not be construed to limit the scope of the invention in any way.
The following Examples are given for the purpose of illustration only, and are not intended to limit the scope of the invention.
Example 1 : Synthesis of 4 ' -O-demethyl-epipodophyllotoxin- /3-D- ( alkenylidene or alkynylidene ) -glucoside derivatives ( Compounds 1-8 ) 200 mg(0.36 mmol) of 4 ' -O-demethyl-epipodophyllotoxin- 3-D-glucoside was dissolved in 20 ml of anhydrous acetonitrile in the presence of 400 mg of 4A molecular sieve and added thereto were 20 mg of p-toluenesulfonic acid and an excess amount ( 2.0-3.5 mmol) of acrolein dimethylacetal (Compound 1), 3-butenaldiethylacetal (Compound 2), 2-butyne-1-aldehyde diethylacetal (Compound 3), phenylpropagylaldehyde( Compound 4), α-methyl-trans- cinnamaldehyde ( Compound 5 ) , 1,2,3, 6-tetrahydrobenz- aldehyde (Compound 6), 3-methyl-2-butenaldehyde( Compound 7) or methacrolein( Compound 8). The resulting mixture was stirred at room temperature under a nitrogen atmosphere for 3 to 24 hours.
After the completion of the reaction was confirmed by TLC analysis, 0.2 ml of triethylamine was added to the reaction mixture to neutralize the acid catalyst. Acetonitrile was removed under a reduced pressure, and then, the residue was dissolved in 200 ml of chloroform. The organic layer was washed with 50 ml of water and dried over anhydrous MgS0A. After filtering off MgSO , the solvent was evaporated under a reduced pressure and the residue was purified by column chromatography to give the following compounds.
Compound 1 : 4 '-0-demethyl-epipodophyllotoxin-/3-D-( 2- propenylidene ) -glucoside
Figure imgf000009_0001
Yield : 72 % p. : 233 - 236 °C
MS : m/z [M]+ = 600
IR(KBr, cm-1) 3451, 2882, 1767, 1609, 1501.
1H NMR (500 MHz, CDClj) 6 6.80(s, IH), 6.52(s, IH), 6.23(s,
2H), 5.98(d, IH), 5.95(d, IH), 5.86(ddd, IH), 5.51(d, IH),
5.40(1H), 5.35(d, IH) , 4.99(d, IH) , 4.90(d, IH), 4.62(d,
IH), 4.58(d, IH), 4.40(dd, IH), 4.24-4.19(m, 2H), 3.73(s,
6H + m, IH), 3.63(m, IH) , 3.43(m, IH) 3.37(m, 2H), 3.25(dd,
IH), 2.87(m, IH), 2.81(d, IH), 2.66(d, IH) .
Compound 2 : 4 '-0-demethyl-epipodophyllotoxin-/3-D-(3- butenylidene) -glucoside;
Figure imgf000010_0001
Yield : 55 % mp. : 210 - 214 °C
MS, m/z [M]+ = 614
IR(KBr, cm-1) 3457, 1771, 1610, 1513, 1483.
1H NMR (500 MHz, CDC13) ό 6.83(s, IH), 6.56(s, IH) , 6.26(s,
2H), 6.01(d, IH), 5.99(d, IH), 5.82(m, IH), 5.46(s, IH),
5.18-5.13(m, 2H), 4.93(d, IH), 4.65-4.60(m, 3H), 4.42(dd,
IH), 4.25-4.20(m, 2H), 3.77(s, 6H), 3.75(m, IH), 3.58(m,
IH), 3.45(t, IH), 3.35-3.27(m, 3H) , 2.89(m, IH), 2.48(m,
2H) . Compound 3 : 4 '-0-demethyl-epipodophyllotoxin-/3-D-(2- butynylidene ) -glucoside;
Figure imgf000011_0001
Yield : 61 %
MS, m/z [M]+ = 612
IR(KBr, cm-1) 3441, 2889, 1768, 1610, 1502. H NMR (500 MHz, CDClj) 6 6.79(s, IH), 6.52 (s, IH) , 6.23(s, 2H), 5.98(d, IH), 5.95(d, IH), 5.41(s, IH), 5.11(dt, IH),
4.89 (d, IH), 4.61(m, IH), 4.57(d, IH), 4.39(t, IH),
4.20(m, 2H), 3.73(s, 6H), 3.70(m, IH), 3.55(m, IH), 3.41(m,
IH), 3.31(m, IH), 3.25(dd, IH), 2.85(m, IH), 1.90(s, 3H) .
Compound 4 : 4 ' -O-demethyl-epipodophyllotoxin-β-D- ( 3- phenyl-2-propynylidene ) -glucoside;
Figure imgf000011_0002
Yield : 52 % mp. : 228 - 231 °C
MS, m/z [M]+ = 674
IR(KBr, cm-1) 3455, 3044, 1768, 1608, 1502.
1H NMR (500 MHz, CDClj) δ 7.52-7.31(m, 5H), 6.83(s, IH),
6.55(s, IH), 6.26(s, 2H), 6.00(d, IH), 5.98(d, IH) , 5.97(s,
IH), 5.43(s, IH), 4.92(d, IH) , 4.67(d, IH), 4.60(d, IH) ,
4.45(dd, IH), 4.23(t, IH), 4.15(t, IH), 4.05(dd, IH) ,
4.00(t, IH), 3.77(m, IH), 3.76(ε, 6H), 3.52-3.37(m, 3H) ,
3.28(dd, IH), 2.90(m, IH) .
Compound 5 : 4 '-O-demethyl-epipodophyllotoxin- β-O- ( 2-methyl-3-phenyl-2-propenylidene) -glucoside;
Figure imgf000012_0001
Yield : 42 % mp. : 216-221 °C MS, m/z M4 = 692 IR(KBr, cm'1) 1769, 1616, 1506, 1231, 1076.
1H NMR (200 MHz, CDC13) : 6 7.23-7.38(m, 5H) , 6.83(s, IH), 6.69(s, IH), 6.55(s, IH) , 6.25(s, 2H), 6.00(d, IH), 5.99(d, IH), 5.47(s, IH), 5.02(s, IH), 4.93(d, IH), 4.66(d, IH) , 4.60(d, IH), 4.20-4.59(m, 3H), 3.76(s, 6H), 3.80-3.86(m, IH), 3.25-3.71(m, 4H), 2.86-2.98(m, 2H) , 1.93(s, 3H) . Compound 6 : 4 '-0-demethyl-epipodophyllotoxin-/3-D- ( 3-cyclohexenylidene) -glucoside;
Figure imgf000013_0001
Yield : 46 % mp. : 207-209 °C
IR(KBr, cm-1) 1769, 1616, 1507, 1231, 1077. H NMR (200 MHz, CDC13) : δ 6.83(s, IH), 6.55(s, IH),
6.25(s, 2H), 6.00(d, IH) , 5.98(d, IH), 5.67(m, 2H) , 5.44(s, IH), 4.91(d, IH), 4.59-4.70(m, 2H), 4.37-4.47(m, IH), 4.18-4.26(m, 2H) , 3.76(s, 6H) , 3.60-3.86(m, IH), 3.22-3.60(m, 4H) , 2.60-2.92(m, 2H) , 1.83-2.06 (m, 7H) .
Compound 7 : 4 ' -0-demethyl-epipodophyllotoxin- 3-D- ( 3- methyl-2-butenylidene ) -glucoside;
Figure imgf000013_0002
Yield : 54 %
IR(KBr, cm-1) 1770, 1616, 1508, 1230, 1037.
1H NMR (200 MHz, CDC13) : δ 6.83(s, IH) , 6.55(s, IH) ,
6.26(s, 2H), 6.00(d, IH), 5.98(d, IH), 5.21-5.28(m, 2H), 4.91(d, IH), 4.67(d, IH) , 4.61(d, IH), 4.18-4.26 ( , 3H),
3.76(ε, 6H), 3.60-3.86(m, IH), 3.21-3.50(m, 4H), 2.75-3.00(m, 2H) , 1.76(s, 6H) .
Compound 8 : 4 '-0-demethyl-epipodophyllotoxin-/3-D-(2- methyl-2-propenylidene) -glucoside;
Figure imgf000014_0001
Yield : 42 % H NMR (200 MHz, CDC13) : δ 6.83(s, IH), 6.55(ε, IH), 6.26(s, 2H), 6.01(d, IH), 5.98(d, IH), 5.21(s. IH) , 5.18(ε, IH), 5.05(s. IH), 4.92(d, IH) , 4.67(d, IH) , 4.61(d, IH) , 4.25-4.23(m, 2H), 3.76(ε, 6H), 1.80(ε, 3H) .
Example 2: Synthesis of 4 '-O-demethyl-epipodophyllotoxin-β- D-(alkoxyalkylidene) -glucoside derivatives (Compounds 9-11)
170 mg(0.30 mmol) of 4 '-O-demethy1-epipodophyllotoxin- 3-D-glucoεide waε dissolved in 20 ml of anhydrous nitromethane and added thereto were 20 mg of ZnCl2 and an excess amount ( 1.8-3.0 mmol) of 3-methoxybutyraldehyde dimethyl acetal (Compound 9), 3-ethyoxypropionaldehyde diethylacetal (Compound 10) or 1, 1, 3-trimethoxypropane (Compound 11) under a nitrogen atmosphere. The resulting mixture was stirred at room temperature for 3 to 24 hours.
After the completion of the reaction was confirmed by TLC analysis, nitromethane was removed under a reduced pressure. The residue was dissolved in 100 ml of chloroform and the organic layer was washed twice with 10 ml portion of water and dried over anhydrous MgS04. After filtering off MgS0A, the solvent was evaporated under a reduced pressure and the residue was purified by column chromatography(eluent: chloroform:methanol = 40:1) to give the following compounds.
Compound 9 : 4 ' -0-demethyl-epipodophyllotoxin-/3-D-( 3- methoxy-l-butylidene)-glucoεide;
Figure imgf000015_0001
Yield : 55 % mp. : 225 - 228 °C
MS, m/z [M]+ = 646
IR(KBr, cm-1) 3424, 2954, 1772, 1610, 1512, 1462. H NMR (500 MHz, CDC13) δ 6.80(ε, IH) , 6.50(ε, IH) , 6.21(ε, 2H), 5.96(s, IH), 5.93(s, IH), 5.48(s, IH) , 4.89(d, IH) , 4.69(m, IH), 4.59-4.55(m, 2H) , 4.38(t, IH) , 4.20-4.14(m, 2H), 3.72(m, IH), 3.72(s, 6H) , 3.64(m, IH), 3.56-3.47(m, 2H), 3.38(m, IH), 3.27(m, IH) , 3.27(s, 3H), 3.12(br. IH), 2.85(m, IH), 1.84(m. IH), 1.69(m, IH), 1.12(d, 3H) .
Compound 10 : 4 '-0-demethyl-epipodophyllotoxin-/3-D-( 3- ethoxy-1-propylidene) -glucoside;
Figure imgf000016_0001
Yield : 62 % mp. : 229 - 231 °C MS, m/z [M]+ = 646
IR(KBr, cm"1) 3442, 2864, 1767, 1609, 1513, 1483. H NMR (500 MHz, CDC13) δ 6.83(s, IH), 6.54(s, IH) , 6.25(s, 2H), 6.00(s, IH), 5.97(8, IH), 5.48(8, IH), 4.92(d, IH), 4.73(t, IH), 4.64-4.58(m, 2H), 4.42(t, IH), 4.25-4.16(m, 2H), 3.75(ε, 6H), 3.70(m, IH), 3.57-3.25(m, 9H) , 2.90(m, IH), 1.96(m, 2H), 1.19(t, 3H) .
Compound 11 : 4 '-0-demethyl-epipodophyllotoxin-/3-D- ( 3- methoxy-1-propy lidene) -glucoside
Figure imgf000016_0002
Yield : 67 % mp. : 206 - 209 °C
MS, m/z [M* + 1] = 633
IR(KBr, cm"1) 3450, 2875, 1767, 1610, 1513, 1483.
1H NMR(500 MHz, CDC13) δ 6.83(s, IH), 6.54(s, IH), 6.25(6, 2H), 6.00(s, IH), 5.97(s, IH), 5.50(s, IH), 4.92(d, IH), 4.72(t, IH), 4.63-4.58(m, 2H), 4.42(t, IH) , 4.25-4.18(m, 2H), 3.75(ε, 6H), 3.73-3.26(m, 8H) , 3.33(s, 3H) , 2.89(m, IH), 1.96(m, 2H).
Example 3: Synthesis of 4 ' -0-demethyl-epipodophyllotoxin-3- D- ( cyclopropylmethylidene ) -glucoside(Compound 12 )
100 mg(0.18 mmol) of 4 '-O-demethyl-epipodophyllotoxin- 3-D-glucoside was dissolved in 10 ml of anhydrous acetonitrile and added thereto were 10 mg of p- tolunesulfonic acid and 0.12 ml of cyclopropane carboxaldehyde. The resulting mixture was stirred at room temperature for 24 hours. After the completion of the reaction was confirmed by TLC analysis, the acid catalyst was neutralized with triethylamine. After removing acetonitrile under a reduced pressure, the resulting solid was disεolved in 20 ml of chloroform. The reεulting solution was washed with 10 ml of a saturated NaCl aqueous solution, and the organic layer was washed twice with 10 ml of portion of water and dried over anhydrous MgSO.. After filtering MgSOA, the solvent was evaporated under a reduced pressure and the residue was subjected to column chromatography(eluent : chloroform: methanol = 40:1) and then column chromatography(eluent : hexane: ethyl acetate = 1:1) to give the title compound.
Figure imgf000017_0001
Yield : 72 % mp. : 229 - 232 °C S, m/z [M]+ = 614
IR(KBr, cm-1) 3448, 2884, 1769, 1609, 1508, 1482. 1H NMR(500 MHz, CDC13) δ 6.80(ε, IH), 6.50(s, IH) , 6.21(s, 2H), 5.97(d, IH), 5.94(d, IH), 5.46(s, IH), 4.88(d, IH), 4.58(d, IH), 4.55(d, IH) , 4.37(dd, IH), 4.20-4.14(m, 3H), 3.72(s, 6H), 3.67(t, IH), 3.50(t, IH), 3.38(t, IH), 3.30-3.23(m, 2H), 3.17(br, IH), 3.00(br, IH) , 2.85(m, IH), 1.12(m, IH), 0.53(m, 2H), 0.45(m, 2H) .
The compound of the present invention thus obtained are shown in Table I .
Table I
Figure imgf000018_0001
Test Example 1 : in vitro anticancer activity
Step 1 : cell cultivation
Cancer cell lines derived from human, i.e., A-549 (non- molding type cell lung cancer), SK-OV-3 (adenocarcinoma, ovary malignant hydroperitoneum) , SK-MEL-2 (malignant melanoma, transferred to femoral skin), XF498(cancer of central nervous system) and HCT15 (colonic cancer) were obtained from U.S. National Cancer Institute and subcultured in Korea Research Institute of Chemical Technology. These cell lines were cultured in a RPMI (Rosewell Park Medium) 1640 culture medium enriched with 5% fetal bovine serum at a constant temperature of 37°C and constant humidity of 98% under an atmosphere of 5% C02. After treating the cultured cells with a phosphate buffer containing 0.25% trypsin and 3 M trans-1, 2-diamino- cyclohexane-N,N,N,N-tetraacetic acid (CDTA) (trypsin-CDTA solution) to separate the cells from the incubator, the cells were subjected to serial subcultures. The serial subcultures were carried out at an interval of three or four days.
Step 2 : in vitro anticancer activity
The in vitro anticancer activity of the compound of the present invention was assessed in accordance with SRB (sulforhodamin B) assay method developed by U.S. National Cancer Institute in 1989.
The subcultured cells were treated with a tripsin-CDTA εolution to separate the cells, which were added to the wells of a 96 well microplate(Falcon Co., U.S.) in an amount of 5 x 103 (A549, HCT15), 1 x 104(SK-MEL-2 , XF498), 2 x 104(SK-OV-3) cells per each wells. The cells were incubated under a C02 atmosphere for 24 hours to be affixed on the well bottom and the culture medium was removed with an aspirator.
The Compounds 1 to 12 of the present invention and etoposide(BMS, U.S.A) were used as test compounds. Each of the test compounds was dissolved in water to obtain a test solution. 0.5% dimethylsulfoxide was used to dissolve the compounds, if necessary. The test solution was diluted with the culture medium(pH 7.2), e.g., by a factor of 1, 10" \ 10"2, 10"3, 10"A and 10-5. After filtering through a 0.22ml filter, 100 ml(x3) of each of the diluted test solutions was added to the wells and the mixture was cultured for another 48 hours.
After removing the culture medium from each well, 100 ml of 10% trichloroacetic acid(TCA) was added to each well and the mixture was stood at 4°C for 1 hour. The plate was washed with water five or six times to completely remove the remaining TCA solution and dried at room temperature.
100 ml of 1% acetic acid solution containing 0.4% sulforhoda in B (SRB) was added to each well. After 30 minutes, the plate was washed with 1% acetic acid solution five or six times to remove unbound SRB and dried at room temperature. Subsequently, 100 ml of an unbuffered 10 mM trisma base solution was added to each well and the plate was shaken by employing a titer plate shaker for 10 minutes to elute the dyeing agent. The absorbance at 520 nm was measured with a icroplate reader.
The anticancer activities (A. . ) of the compounds were calculated by equation (1) or (2).
A. A. = [ (T-Tz)/(C-Tz) ] X 100 (Tz>T) Eq. (1) A. A. = [(T-Tz)/Tz] x 100 (Tz<T) Eq. (2)
wherein, Tz is the number of cells before adding a test compound without incubation; C is the number of cells after incubation for 48 hours in the absence of a test compound; and T is the number of cells after incubation for 48 hours the cell in the presence of a test compound.
From the values thus obtained, ED50, i.e., the concentration of a compound to inhibit the growth of a cancer cell by 50% was calculated by regression method using LOTUS program.
The results are shown in Table II.
Test Example 2 : Inhibition of human topoisomeraεe II
Step 1 : Preparation of topoisomerase II HeLa cell line provided by ATCC(American Type Culture Collection) was used in preparing topoiso erase II of the cancer cell.
The HeLa cells were cultured in a RPMI 1640 culture medium enriched with 10% fetal bovine serum at 37°C under an atmosphere of 5% C02 and the cultured HeLa cells were collected. Added thereto was a 3-fold volume of 50 mM KH2POA buffer solution(pH 7.0) containing 1 mM ethylenediamine tetraacetic acid(EDTA), ImM mercaptoethanol, 0.5 mM phenylmethylsulfonylchloride(PMSF) and 10% glycerol and the resulting mixture was homogenized with Polytron* for 30 seconds in ice bath.
A 2M KH2PO buffer solution(pH 7.0) was added to a KH2PO concentration of 0.3M and the resultant was homogenized for 30 seconds, placed in an ice bath for 1 hour, and then, centrifuged at 30,000 xg for 30 minutes. The supernatant was purified by phosphocellulose column chromatography(eluent : linear gradient from 0.3M to 0.5M KH2P04 buffer(pH 7.0) to obtain topoisomerase II.
Step 2 : P unknotting assay
IC50 values of the test compounds for topoisomerase II (Topo II) were measured by an unknotting assay. Topo II obtained above and 0.4 μg of P knotted DNA were added to a 50 mM N-[ 2-hydroxyethyl]-piperazine-N'-[ 2- ethanesulfonic acid] (HEPES) buffer solution (pH 7.0) containing 50 mM KC1, 0.1 mM EDTA, 100 mM NaCl, 10 mM MgCl2, 100 μg/ml of fetal bovine serum albumin and 1 mM adenosine triphosphate(ATP) to a final volume of 20 μl and the mixture waε reacted at 37 °C for 30 minutes.
The reaction was terminated by adding sodium dodecyl sulfate(SDS) to a SDS concentration of 1%. The resultant was subjected to electrophoresis using 1% agarose gel equilibrated with a 50 mM tris borate buffer solution(pH 8.3) containing 2.5mM EDTA, and then, dyed with ethidium bromide .
The activity of Topo II was assessed using photographs taken under UV radiation. The Topo II activity measured when 50% of 0.4 μg of knotted PA DNA was unknotted was defined as 1 unit activity. IC50 was the concentration of a compound to inhibit the activity of Topo II by 50%. The results are shown in Table II.
Table II.
Figure imgf000022_0001
As shown in Table II, some of the compounds of the present invention exhibit superior or comparable anticancer activities against the five cancer cells listed in Table II in comparison with etoposide.
Test Example 3 : In. vivo anticancer activity
6-Week old BDF1 mice were divided into seven groups (n=8), one control group and six test groups. A dose of 1 x 10s cells/0. lml of leukemia L1210 cell subcultured in DBA/2 mouse was injected into the abdominal cavity of each of the mice. Two test compoundε, etoposide and Compound 1 of the present invention, were injected into the abdominal cavities of mice of respective test group in the doses shown in Table III, successively at 1st, 2nd, 3rd, 4th and 5th days from the leukemia cell injection.
The mice were observed daily and the survival period of each mouse was checked. The anticancer activity was assessed by calculating the ratio (I.R.) by equation 3.
I.R. = 100 x (At - Ac)/Ac Eq (3) wherein, At is the average survival days of mice in a test group and Ac is the average survival days of mice in the contro1 group.
The results are shown in Table III.
Table III
Figure imgf000023_0001
As the above results show, the novel 4 '-O-demethy1- epipodophyllotoxin-/3-D-glucoside derivative of formula(I) of the present invention possesses an excellent anticancer activity against various cancer cells, much higher than that of etoposide. The inventive compounds can therefore be used as a drug for treating cancers.
While the invention has been described with respect to the specific embodiments, it should be recognized that various modifications and changes may be made by those skilled in the art to the invention which also fall within the scope of the invention aε defined by the appended claims.

Claims

What is claimed is :
1. A 4 '-0-demethyl-epipodophyllotoxin-/3-D-glucoside acetal derivative of formula (I):
R'
Figure imgf000024_0001
wherein
R1 and R2 are, each independently, hydrogen, a C2_12 alkenyl, C2_12 alkynyl, C2_12 alkoxylalkyl or cyclopropyl group optionally sub╬╡tituted with one or more radicals selected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups, with the proviso that R1 and R2 are not hydrogen simultaneously; or R1 and R2, together with the carbon atom to which they are attached, form an unsaturated C5_15 ring optionally substituted with one or more radicals ╬╡elected from the group consisting of alkoxy, phenyl, substituted phenyl, halogen, nitro, cyano, hydroxy, carboxyl and amino groups.
2. The compound of claim 1 , wherein R is hydrogen and R2 is a C2_12 alkenyl, C2_12 alkynyl, C2_12 alkoxylalkyl or cyclopropyl group optionally substituted with phenyl or substituted phenyl group.
3. The compound of claim 2, which is selected from the group consi╬╡ting of:
4 ' -0-demethyl-epipodophyllotoxin-/3-D- ( 2-propenylidene ) - glucoside;
4 ' -0-demethyl-epipodophyllotoxin-3-D- ( 3-butenylidene ) - glucoside;
4 ' -0-demethyl-epipodophyllotoxin-3-D- ( 2-butynylidene ) - glucoside;
4'-0-demethyl-epipodophyllotoxin-/3-D-(3-phenyl-2- propynylidene)-glucoside;
4 '-0-demethyl-epipodophyllotoxin-3-D-(2-methyl-3-phenyl-2- propeny lidene ) -glucoside; 4 ' -O-demethyl-epipodophyllotoxin-jS-D- ( 3-cyclohexenylidene ) - glucoside;
4 '-0-demethyl-epipodophyllotoxin-3-D-(3-methyl-2- buteny lidene ) -glucoside ;
4'-0-demethyl-epipodophyllotoxin-/3-D-(2-methyl-2- propenylidene) -glucoside;
4 ' -O-demethy 1 -epipodophy llotoxin-/3-D- ( 3-methoxy-l- buty lidene ) -glucoside ;
4'-0-demethyl-epipodophyllotoxin-/3-D-( 3-ethoxy-l- propylidene) -glucoside; 4 ' -O-demethy 1 -epipodophy llo toxin-3-D- (3-methoxy-l- propylidene) -glucoside; and
4 ' - 0- demethy l - ep ipodophy l l otox in -/3 - D -
( cyclopropylmethylidene ) -glucoside .
4. A process for preparing the compound of claim 1 which comprises reacting 4 '-O-demethyl-epipodophyllotoxin- /3-D-glucoside of formula (II) with an aldehyde of formula(III) or an acetal of formula(IV) in the presence of a catalyst:
Figure imgf000025_0001
Figure imgf000026_0001
Figure imgf000026_0002
wherein, R1 and R2 have the ╬╡ame meanings as defined above, and R3 is a C,_3 alkyl group.
5. An anticancer composition comprising a therapeutically effective amount of one or more of the 4'-
0-demethyl-epipodophyllotoxin-3-D- glucoside acetal derivatives of claim 1 as an active ingredient, and a pharmaceutically acceptable carrier and/or adjuvant.
PCT/KR1998/000452 1997-12-23 1998-12-22 4'-O-DEMETHYL-EPIPODOPHYLLOTOXIN-β-D-GLUCOSIDE ACETAL DERIVATIVES WO1999032499A1 (en)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3524844A (en) * 1965-09-21 1970-08-18 Sandoz Ag Epipodophyllotoxin glucoside derivatives
EP0196618A1 (en) * 1985-04-02 1986-10-08 Microbial Chemistry Research Foundation 4'-Demethyl-4-epipodophyllotoxin Derivatives, a process for their preparation and their use as medicaments
GB2207674A (en) * 1987-08-04 1989-02-08 Bristol Myers Co Podophyllotoxin derivatives
EP0367189A2 (en) * 1988-10-31 1990-05-09 Bristol-Myers Squibb Company Sugar phosphates and sulfonates of epipodophyllotoxin glucosides

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3524844A (en) * 1965-09-21 1970-08-18 Sandoz Ag Epipodophyllotoxin glucoside derivatives
EP0196618A1 (en) * 1985-04-02 1986-10-08 Microbial Chemistry Research Foundation 4'-Demethyl-4-epipodophyllotoxin Derivatives, a process for their preparation and their use as medicaments
GB2207674A (en) * 1987-08-04 1989-02-08 Bristol Myers Co Podophyllotoxin derivatives
EP0367189A2 (en) * 1988-10-31 1990-05-09 Bristol-Myers Squibb Company Sugar phosphates and sulfonates of epipodophyllotoxin glucosides

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