WO2002081491A2 - New genistein derivatives and pharmaceutical preparations containing them - Google Patents

New genistein derivatives and pharmaceutical preparations containing them Download PDF

Info

Publication number
WO2002081491A2
WO2002081491A2 PCT/PL2002/000029 PL0200029W WO02081491A2 WO 2002081491 A2 WO2002081491 A2 WO 2002081491A2 PL 0200029 W PL0200029 W PL 0200029W WO 02081491 A2 WO02081491 A2 WO 02081491A2
Authority
WO
WIPO (PCT)
Prior art keywords
group
genistein
hydrogen atom
alkyl
aryl
Prior art date
Application number
PCT/PL2002/000029
Other languages
French (fr)
Other versions
WO2002081491A3 (en
Inventor
Osman Achmatowicz
Grzegorz Grynkiewicz
Wieslaw Pucko
Aleksander P. Mazurek
Krzysztof Polkowski
Jerzy Boryski
Wieslaw Szeja
Wieslaw Szelejewski
Adam Opolski
Gabriella Pastuch-Gawolek
Czeslaw Radzikowski
Barbara Szechner
Joanna Wietrzyk
Janusz Skierski
Ilona Wandzik
Piotr Krzeczynski
Original Assignee
Instytut Farmaceutyczny
Instytut Leków
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Instytut Farmaceutyczny, Instytut Leków filed Critical Instytut Farmaceutyczny
Publication of WO2002081491A2 publication Critical patent/WO2002081491A2/en
Publication of WO2002081491A3 publication Critical patent/WO2002081491A3/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/26—Acyclic or carbocyclic radicals, substituted by hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
    • C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
    • C07D311/34—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 3 only
    • C07D311/36—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 3 only not hydrogenated in the hetero ring, e.g. isoflavones
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
    • C07F7/02—Silicon compounds
    • C07F7/08—Compounds having one or more C—Si linkages
    • C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
    • C07F7/1804—Compounds having Si-O-C linkages

Definitions

  • the present invention relates to the new genistein derivatives and pharmaceutical preparations containing them.
  • New genistein derivatives exhibit antitumour activity.
  • Natural isoflavones especially genistein (5,7,4'-trihydroxy-3- phenylchromen-4-on) are characterised by multidirectional biological activity, especially by ability to inhibit protein tyrosine kinases activity - the group of enzymes of significant importance for protooncogens and oncogens expression. This activity influences potential use of genistein in therapy and prevention of neoplastic diseases, circulatory diseases, osteoporosis and other diseases. The total amount of isoflavones in plant raw materials is small.
  • soy products contain from 0.1 to 0.3% of various components which in case of Glycine max Merill seeds (soy beans) are ⁇ -D-glucosides: genistin, daidzin, glycitin.
  • the aglycones corresponding to them are practically not present in the raw material and appear in significant amount only after thermal processing or fermentation. Due to a suggestion of researchers that glycosides of isoflavones, and not their aglycones are responsible for advantageous action of soy, interest in synthetic preparation of glycosides and other genistein derivatives is growing.
  • the present invention provides a number of genistein derivatives containing selectively introduced certain functional groups which may play important biological roles, especially they alter the pathways by which genistein may be eliminated or transformed as a result of metabolic processes in the organism.
  • the invention resides provides the new genistein derivatives of formula 1, wherein: R ⁇ and R 2 are the same or different and independently represent:
  • R 5 , R 6 and R 7 are the same or different and denote C ⁇ -6 alkyl or aryl, - mono-, di- or oligosaccharide group, whereby at least one hydroxyl group of saccharide group may be substituted by the same or different acyl, alkyl, acyloxyalkyl or aryl groups;
  • R 3 denotes hydrogen atom or -COCH 3 group;
  • Ri denotes hydrogen atom, group -SO 3 H, SO 3 " , -NH 2 or -NO 2 groups.
  • alkyl should be understood as a straight or branched hydrocarbon chain containing from 1 to 18 carbon atoms and, eventually, one or more unsaturated bonds excluding chains containing 17 carbon atoms corresponding to residues of oleinic and stearic acids.
  • aryl should be understood as cyclic unsaturated group containing at least four carbon atoms and, eventually, one or more heteroatoms chosen out of nitrogen, oxygen and sulfur.
  • saccharide group should be understood as a sugar derivative: pentose, hexose or heptose in a chain or cyclic form.
  • the preferable genistein derivatives are expressed by formula 1, wherein R ⁇ represent alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl, R 5 (R 6 )R 7 -Si- group, R 2 denotes hydrogen atom or group -COCH 3 , an R 3 and R_ t are simultaneously hydrogen atom.
  • the preferable genistein derivatives are expressed by formula 1, wherein Rjand R t are hydrogen atom, R 3 is hydrogen atom or -COCH 3 group, and R 2 represents alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl or R 5 (R 6 )R 7 -Si- group.
  • the preferable genistein derivatives are expressed by formula 1, wherein Ri and R are the same or different and denote alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl or R 5 (R 6 )R 7 -Si-, and R 3 is hydrogen atom or -COCH 3 , and R t is hydrogen atom.
  • the preferable group of compounds according to the invention are genistein glycosides expressed by formula 1, wherein R ls R 3 and R 4 are hydrogen atom, and R 2 is saccharide group.
  • glycosides are furanosides.
  • Genistein derivatives according to the invention exhibit a potent and selective antiproliferative and cytotoxic action and some of them, in addition, for example silyl derivatives, may be valuable intermediates for preparation of other derivatives.
  • Genistein derivatives expressed by formula 1 may be prepared, depending upon a kind and a place of a substituent position, by use of genistein, its synthetic precursor - 2,4,6,4'-tetradeoxybenzoin or genistin (7-O- ⁇ -D- glucopyranosylgenistein) as a substrate.
  • Si-, and R 3 and Rt are hydrogen atom.
  • Mono-O-acyl derivatives of genistein have conveniently diversified functional groups: base-labile 7-O-acyl group, acid-labile 4-O-alkylsilyl group and non-reactive hydroxyl group in position 5, "protected" by intramolecular hydrogen bond.
  • Such derivatives may serve (after partial deprotection) as substrates for another genistein derivatives preparation, which contain two different substituents in positions 4'- and 7-.
  • Diacyl genistein derivatives expressed by formula 1, wherein Ri and R 2 are the same and represent alkylcarbonyl or arylcarbonyl, and R 3 and Rt are hydrogen atom, are prepared by reaction of genistein with at least 4-fold molar excess of acylating agent, such as acid chloride or acid anhydride in the presence of a base.
  • acylating agent such as acid chloride or acid anhydride
  • Suitable bases are pyridine, substituted pyridines, trialkylamines and cyclic tertiary amines.
  • Genistein derivatives expressed by formula 1, wherein R l5 R 3 and R 4 are hydrogen atom, R 2 represents alkyl, allyl or arylalkyl, are prepared by known in the art method consisting in the reaction of genistein with a slight molar excess of alkylating agent, such as alkyl halide in the presence of base or by generating genistein phenoxide anion through the reaction with a potent base or metal hydride first, and then by the reaction with alkylating agent.
  • alkylating agent such as alkyl halide
  • genistein derivatives containing an alkyl substituent in position 5 are prepared by monoalkylation of tetradeoxybenzoin, which is then subjected to a formylation and cyclization reaction, similarly as described in the Polish Patent application pending No.343505 for genistein preparation.
  • genistein derivatives substituted in position 3' expressed by the general formula 1, wherein Rt represents SO 3 H, SO 3 " , -NH 2 or -NO 2 group, are prepared by typical reactions of sulfonation, nitration or stepwise introduction of an amino group applying genistein as a substrate.
  • Genistein derivatives expressed by formula 1, wherein Ri denotes hydrogen atom, R 2 denotes saccharide group, and R 3 and Rt are hydrogen atom, may be prepared by one, known in the chemistry of sugars, method of creating glycoside bond in the reaction of glycosyl donor with glycosyl acceptor in the presence of promoter, preferably in organic solvent, by use of thermal or microwave activation.
  • 1 -glycosyl derivatives are applied as glycosyl donors, such as halides, esters, ethers or glycals in which anomeric substituent may be transformed in a good leaving group.
  • glycosidation reaction may be carried out in acidic conditions in the presence of Lewis' or Br ⁇ nsted's acids or, in the case of anomeric exchange reaction, in basic conditions.
  • Lewis' or Br ⁇ nsted's acids are, for example, toluenesulfonic or trifluoromethylsulfonic acids, and proper Lewis' acids are boron trifluoride, tin tetrachloride, aluminium trichloride or titanium tetrachloride.
  • Genistein glycosides according to the invention are preferably prepared by anomeric exchange reaction known, for example, from publication in Nucleosides, Nucleotides, 15, 771 (1996).
  • As a glycosyl acceptor genistein or its acyl, alkyl or silyl derivatives are used. Use of selectively functionalized derivatives is preferable due to their better solubility in reaction medium.
  • This reaction exhibits unexpected regioselectivity of glycosidation reaction, allowing for preparation of genistein glycosides substituted in position 4', as main reaction products.
  • Glycosidation reaction is carried out in anhydrous conditions, preferably in neutral gas atmosphere, in aprotic solvent or their mixture, at elevated temperature. As a result of glycosidation, a mixture of anomers of genistein glycosides is obtained which, if needed, may be separated by gel chromatography or crystallization.
  • the obtained glycoside derivatives of genistein may be further selectively functionalised in position 5" of a pentofuranose or 6" of hexopyranose in the saccharide group by use of acylating, alkylating, silylating or glycosidating reagents.
  • the hydroxyl group in position 5" may be selectively protected by the method known in chemistry of sugars in the form of esters of mono- or dicarboxylic acids, orthoesters, carbamates, sulfonates, phosphates and ethers, for example: trityl, silyl, alcoxyalkyl o tetrahydropyranyl.
  • Genistein derivatives according to the invention are characterized by various physicochemical properties while maintaining the desired biological activity of the parent compound.
  • Selective functionalising of genistein allows obtaining compounds of desired properties, such as better solubility in body fluids, affinity to biopolymers, susceptibility to degradation, lipophilicity (affinity to cell membranes) which, in turn, influence pharmacokinetic and pharmacodynamic properties of the compound.
  • desired properties such as better solubility in body fluids, affinity to biopolymers, susceptibility to degradation, lipophilicity (affinity to cell membranes) which, in turn, influence pharmacokinetic and pharmacodynamic properties of the compound.
  • 4'-furanosides and 7-pyranosides of genistein and their ester and ether derivatives show multidirectional biological activity, both as direct precursors of genistein of high bioavailability and as integral ligands of functionally important endogenous biopolymers, so the range of possible biological activity and medical applications of new derivatives includes all actions of genistein, but is not limited by them.
  • Genistein derivatives according to the invention were examined from the point of view of biological activity in selected tumour cell lines in various tests of cytotoxicity and cytostatics using statistical cytometry (SC) and flow cytometry (FC) techniques.
  • SC statistical cytometry
  • FC flow cytometry
  • the potency of toxic activity (relative cytotoxicity) of the tested compounds was expressed with regard to genistein, as a ratio of percentage of alive cells in a culture with genistein to percentage of alive cells in a culture with the tested compound.
  • the influence of the examined compounds on the rate of proliferation was expressed with regard to genistein as a ratio of cell culture density in the presence of genistein to cell density in the presence of the tested compound.
  • the invention includes the pharmaceutical preparation containing the active substance which is the genistein derivative of formula 1, wherein Ri and R 2 are the same or different and independently represent:
  • R 5 , R 6 and R 7 are the same or different and denote alkyl C 1-6 or aryl, - mono-, di- or oligosaccharide group, while at least one hydroxyl group of saccharide group may be substituted by the same or different acyl, alkyl, acyloxyalkyl or aryl groups;
  • R 3 denotes hydrogen atom or -COCH 3 group, and
  • Ri denotes hydrogen atom, - SO 3 H, SO 3 " , -NH 2 or -NO 2 group or its pharmaceutically acceptable salt, together with the usual carriers and/or auxiliaries.
  • the preparation according to the invention may be in a pharmaceutical dosage form suitable for oral, parenteral, intranasal, sublingual or rectal use or for administration by inhalation or insufflation.
  • Especially the preparation may be in the dosage form of a tablet, pill, capsule, powder, granules, sterile solution or suspension, aerosol, ampoule, liposomal preparation or suppository.
  • Solid dosage forms such as tablets, pills, powders, granules or capsules are prepared by mixing the active substance with the pharmaceutically acceptable carrier, such as corn starch, lactose, saccharose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums and other pharmaceutical diluents, for example water, in order to obtain solid preliminary mixture containing homogenous mixture of the compound according to the invention or its pharmaceutically acceptable salt.
  • the mixture obtained in this way may be then tabletted or drageetted or the capsules may be filled with it.
  • Tablets or granules of the new composition may be coated or processed in another way in order to obtain unit form ensuring preferable prolonged action.
  • a range of various substances may be used for preparation of such protective or coating layers including various polymeric acids and mixtures of polymeric acids with such substances as shellac, cetyl alcohol or cellulose acetate.
  • Liquid dosage forms appropriate for oral or parenteral administration of the preparations according to the invention include water solutions, syrups, water or oil suspensions, emulsions with edible oils such as cotton seeds oil, sesame oil, coconut or peanut oil as well as elixirs with similar pharmaceutical carriers.
  • Appropriate dispergating or suspending agents for water suspensions include synthetic or natural gums such as tragacanth, acacia, alginate, dextran, carboxymethyl sodium cellulose, methylcellulose, polyvinyl pyrrolidone or gelatine.
  • compositions according to the invention may be used in chemotherapy and/or chemoprevention of neoplasms.
  • Example 2 7-tert-Butyldimethylsilyloxy-5-hydroxy-3-(4'-t- butyldimethylsilyloxyphenyl)chromen-4-on (3)
  • imidazole (2.72 g, 40 mmol) and tert-butyldimethylsilyl chloride (3.3 g, 22 mmol) were added to genistein solution 1 (2.7 g, 10 mmol) in DMF (30ml). The reaction was carried out at room temperature for 1.5 hours. 5.34 g of yellow precipitate was obtained, it was purified by chromatography with eluting mixture of hexamethyl acetate (20:1 v/v).
  • Acetic anhydride (1.5ml, 16 mmol) was added to the solution of compound 3 (4g, 8 mmol) (obtained according to the example 2) in anhydrous pyridine (15 ml). The reaction was carried out at room temperature for 3 hours. After that time the mixture was diluted with toluene (100 ml) and rinsed with cold water (3 x 30 ml). Toluene solution was dried with anhydrous MgSO 4 , concentrated under vacuum on the evaporator and additionally evaporated with toluene (2 x 10 ml). The obtained yellow solid (4.03 g) was purified on chromatographic column with use of eluting mixture of hexamethyl acetate (15:1 vol). 2.32g (yield: 62%) of product 4 in the form of white crystals and 1.02 g (yield: 30%) of product 5 in the form of light yellow crystals was obtained.
  • Genistein solution (1) (270mg, 1 mmol) in DMF (3 ml) with addition of diisopropylethylamine (DIPEA) (155 mg, 1.2 mmol) was stirred at room temperature for 20 minutes. After that time, pivaloyloxymethyl chloride (180, 1.2 M) and catalytic amount of 4-dimethylaminopyridine (DMAP) were added to homogenous mixture. The mixture was stirred at room temperature for 5 days.
  • DIPEA diisopropylethylamine
  • DMAP 4-dimethylaminopyridine
  • Catalytic amount of SnCl 4 (333mg,1.28 mmol) was added to the mixture of genistein (1) (270 mg, 1 mmol), 2,3,5-tri-O-benzylarabinofuranose(428 mg, 2 mmol) in anhydrous acetonitrile (5 ml) cooled to °C. Then the mixture was stirred at room temperature for 40 minutes. After that time the mixture was poured on water with ice (30 g) with addition of saturated solution of NaHCO 3 (20 ml), the neutral mixture was rinsed with ethyl acetate (50 ml) and water.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Saccharide Compounds (AREA)

Abstract

Genistein derivatives of Formula (1), wherein R1 and R2 are the same or different and independently represent hydrogen atom, alkyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl, while each of the above mentioned groups may be substituted in a chain or ring by amino, nitro or nitrile groups, R5(R6)R7-Si- group wherein R5, R6 and R7 are the same or different and denote C1-6alkyl or aryl, mono-, di- or oligosaccharide group while at least one hydroxyl group of saccharide group may be substituted by the same or different acyl, alkyl, acyloxyalkyl or aryl groups; R3 represents hydrogen atom or -COCH3 group; and R4 represents hydrogen atom, -SO3H, SO3- or -NH¿2? or -NO2 group; and their pharmaceutically acceptable salts exhibit antyproliferative and antitumour activity. New derivatives and pharmaceutical preparations containing them may be suitable in clinical oncology and/or chemoprevention of neoplasms.

Description

New genistein derivatives and pharmaceutical preparations containing them
The present invention relates to the new genistein derivatives and pharmaceutical preparations containing them. New genistein derivatives exhibit antitumour activity.
Natural isoflavones, especially genistein (5,7,4'-trihydroxy-3- phenylchromen-4-on) are characterised by multidirectional biological activity, especially by ability to inhibit protein tyrosine kinases activity - the group of enzymes of significant importance for protooncogens and oncogens expression. This activity influences potential use of genistein in therapy and prevention of neoplastic diseases, circulatory diseases, osteoporosis and other diseases. The total amount of isoflavones in plant raw materials is small. For example, different soy products contain from 0.1 to 0.3% of various components which in case of Glycine max Merill seeds (soy beans) are β-D-glucosides: genistin, daidzin, glycitin. The aglycones corresponding to them (genistein, daidzein and glycitein) are practically not present in the raw material and appear in significant amount only after thermal processing or fermentation. Due to a suggestion of researchers that glycosides of isoflavones, and not their aglycones are responsible for advantageous action of soy, interest in synthetic preparation of glycosides and other genistein derivatives is growing.
Recent reports on genistin (7-O-β-D-glucopyranosylgenistein) preparation (7-O-β-D-glucopyranosylgenistein) indicate, that hydroxy groups of genistein are not equivalent regarding their chemical reactivity and that 7-hydroxyl group shows ca. 100 times greater acidity than 4'-hydroxyl group. The expected (on the basis of calculation of acidity) sequence of substitution of hydrogen atoms in phenol groups (7-OH»4'-OH>5-OH) is, however, not reflected in known structures of simple genistein derivatives that are usually a result of exhaustive alkylation or acylation (for example, there are tris-trimethylsilyl, trimethyl, triethyl, triacetyl and tribenzoyl derivatives known).
Recently, in Tetrahedron 56(2000), 7805 publication, mono- and diesters of genistein with oleinic and stearic acids were described along with regioselective method of their preparation.
The present invention provides a number of genistein derivatives containing selectively introduced certain functional groups which may play important biological roles, especially they alter the pathways by which genistein may be eliminated or transformed as a result of metabolic processes in the organism.
The invention resides provides the new genistein derivatives of formula 1, wherein: R\ and R2 are the same or different and independently represent:
- hydrogen atom - alkyl
- allyl
- aryl
- alkyloaryl
- alkylcarbonyl - arylcarbonyl, while each of the above mentioned groups may be substituted in a chain or ring by amino, nitro or nitrile groups,
- R5(R6)R7-Si- group, wherein R5, R6 and R7 are the same or different and denote Cι-6alkyl or aryl, - mono-, di- or oligosaccharide group, whereby at least one hydroxyl group of saccharide group may be substituted by the same or different acyl, alkyl, acyloxyalkyl or aryl groups; R3 denotes hydrogen atom or -COCH3 group; and Ri denotes hydrogen atom, group -SO3H, SO3 ", -NH2 or -NO2 groups. The term "alkyl" should be understood as a straight or branched hydrocarbon chain containing from 1 to 18 carbon atoms and, eventually, one or more unsaturated bonds excluding chains containing 17 carbon atoms corresponding to residues of oleinic and stearic acids. The term "aryl" should be understood as cyclic unsaturated group containing at least four carbon atoms and, eventually, one or more heteroatoms chosen out of nitrogen, oxygen and sulfur.
The term "saccharide group" should be understood as a sugar derivative: pentose, hexose or heptose in a chain or cyclic form.
In the first embodiment of the invention, the preferable genistein derivatives are expressed by formula 1, wherein R} represent alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl, R5(R6)R7-Si- group, R2 denotes hydrogen atom or group -COCH3, an R3 and R_t are simultaneously hydrogen atom. In the second embodiment of the invention, the preferable genistein derivatives, according to the invention, are expressed by formula 1, wherein Rjand Rt are hydrogen atom, R3 is hydrogen atom or -COCH3 group, and R2 represents alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl or R5(R6)R7-Si- group.
In another embodiment of the invention, the preferable genistein derivatives, according to the invention, are expressed by formula 1, wherein Ri and R are the same or different and denote alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl or R5(R6)R7-Si-, and R3 is hydrogen atom or -COCH3, and Rt is hydrogen atom.
The preferable group of compounds according to the invention are genistein glycosides expressed by formula 1, wherein Rls R3 and R4 are hydrogen atom, and R2 is saccharide group.
Especially preferable glycosides are furanosides.
Genistein derivatives according to the invention exhibit a potent and selective antiproliferative and cytotoxic action and some of them, in addition, for example silyl derivatives, may be valuable intermediates for preparation of other derivatives.
Genistein derivatives expressed by formula 1 may be prepared, depending upon a kind and a place of a substituent position, by use of genistein, its synthetic precursor - 2,4,6,4'-tetradeoxybenzoin or genistin (7-O-β-D- glucopyranosylgenistein) as a substrate.
Genistein with hydroxyl groups in positions 7 and 4' protected in the form silyl derivatives, expressed by formula 1, wherein R\ and R2 simultaneously represents R5(R6)R7-Si-, and R3 and R4 are hydrogen atom, is prepared by reaction of genistein with a molar excess of alkylsilyl chloride in the presence of imidazole, in organic solvent such as dimethylformamide. Disilyl derivatives obtained by this way may be then subjected to the reaction of selective acylation in position 7, by use of proper chloride or acid anhydride in the presence of organic base, such as pyridine. Mono-O-acyl derivatives of genistein of formula 1 are obtained in this way, wherein R\ represents alkylcarbonyl or arylcarbonyl, R2 represents R5(R6)R7-
Si-, and R3 and Rt are hydrogen atom.
Mono-O-acyl derivatives of genistein have conveniently diversified functional groups: base-labile 7-O-acyl group, acid-labile 4-O-alkylsilyl group and non-reactive hydroxyl group in position 5, "protected" by intramolecular hydrogen bond. Such derivatives may serve (after partial deprotection) as substrates for another genistein derivatives preparation, which contain two different substituents in positions 4'- and 7-.
7-Mono-O-acyl genistein derivatives expressed by formula 1, wherein R denotes alkylcarbonyl or arylcarbonyl, and R2, R3 and Rt are hydrogen atom, are obtained from disilyl compounds by removing silyl protective group from position
4' by reaction with diluted acids or reagents liberating fluoride anion..
Diacyl genistein derivatives expressed by formula 1, wherein Ri and R2 are the same and represent alkylcarbonyl or arylcarbonyl, and R3 and Rt are hydrogen atom, are prepared by reaction of genistein with at least 4-fold molar excess of acylating agent, such as acid chloride or acid anhydride in the presence of a base.
Suitable bases are pyridine, substituted pyridines, trialkylamines and cyclic tertiary amines.
In the reaction of genistein with slight molar excess of acylating agent (1.2-1.8 M for 1 M of genistein), the mixture of 7-O-acetyl and 4'-O-acetyl derivatives of genistein is obtained, which may be then separated by chromatography method.
Genistein derivatives expressed by formula 1, wherein Rl5 R3 and R4 are hydrogen atom, R2 represents alkyl, allyl or arylalkyl, are prepared by known in the art method consisting in the reaction of genistein with a slight molar excess of alkylating agent, such as alkyl halide in the presence of base or by generating genistein phenoxide anion through the reaction with a potent base or metal hydride first, and then by the reaction with alkylating agent. Next, genistein derivatives containing an alkyl substituent in position 5 are prepared by monoalkylation of tetradeoxybenzoin, which is then subjected to a formylation and cyclization reaction, similarly as described in the Polish Patent application pending No.343505 for genistein preparation. Finally, genistein derivatives substituted in position 3' expressed by the general formula 1, wherein Rt represents SO3H, SO3 ", -NH2 or -NO2 group, are prepared by typical reactions of sulfonation, nitration or stepwise introduction of an amino group applying genistein as a substrate.
Genistein derivatives expressed by formula 1, wherein Ri denotes hydrogen atom, R2 denotes saccharide group, and R3 and Rt are hydrogen atom, may be prepared by one, known in the chemistry of sugars, method of creating glycoside bond in the reaction of glycosyl donor with glycosyl acceptor in the presence of promoter, preferably in organic solvent, by use of thermal or microwave activation. In the methods widely known in the art, 1 -glycosyl derivatives are applied as glycosyl donors, such as halides, esters, ethers or glycals in which anomeric substituent may be transformed in a good leaving group. Depending upon a type of anomeric substituent, glycosidation reaction may be carried out in acidic conditions in the presence of Lewis' or Brδnsted's acids or, in the case of anomeric exchange reaction, in basic conditions. Proper Bronsted's acids are, for example, toluenesulfonic or trifluoromethylsulfonic acids, and proper Lewis' acids are boron trifluoride, tin tetrachloride, aluminium trichloride or titanium tetrachloride.
Genistein glycosides according to the invention are preferably prepared by anomeric exchange reaction known, for example, from publication in Nucleosides, Nucleotides, 15, 771 (1996). As a glycosyl acceptor, genistein or its acyl, alkyl or silyl derivatives are used. Use of selectively functionalized derivatives is preferable due to their better solubility in reaction medium. This reaction exhibits unexpected regioselectivity of glycosidation reaction, allowing for preparation of genistein glycosides substituted in position 4', as main reaction products. Glycosidation reaction is carried out in anhydrous conditions, preferably in neutral gas atmosphere, in aprotic solvent or their mixture, at elevated temperature. As a result of glycosidation, a mixture of anomers of genistein glycosides is obtained which, if needed, may be separated by gel chromatography or crystallization.
The obtained glycoside derivatives of genistein may be further selectively functionalised in position 5" of a pentofuranose or 6" of hexopyranose in the saccharide group by use of acylating, alkylating, silylating or glycosidating reagents. Especially, the hydroxyl group in position 5" may be selectively protected by the method known in chemistry of sugars in the form of esters of mono- or dicarboxylic acids, orthoesters, carbamates, sulfonates, phosphates and ethers, for example: trityl, silyl, alcoxyalkyl o tetrahydropyranyl.
Genistein derivatives according to the invention are characterized by various physicochemical properties while maintaining the desired biological activity of the parent compound.
Selective functionalising of genistein allows obtaining compounds of desired properties, such as better solubility in body fluids, affinity to biopolymers, susceptibility to degradation, lipophilicity (affinity to cell membranes) which, in turn, influence pharmacokinetic and pharmacodynamic properties of the compound. Especially 4'-furanosides and 7-pyranosides of genistein and their ester and ether derivatives show multidirectional biological activity, both as direct precursors of genistein of high bioavailability and as integral ligands of functionally important endogenous biopolymers, so the range of possible biological activity and medical applications of new derivatives includes all actions of genistein, but is not limited by them.
Genistein derivatives according to the invention were examined from the point of view of biological activity in selected tumour cell lines in various tests of cytotoxicity and cytostatics using statistical cytometry (SC) and flow cytometry (FC) techniques.
Methods Examination of antitumour activity of genistein derivatives was carried out by evaluation of influence of these compounds on survival rate and proliferation in vitro. Leukemic cell lines were used in experiments: Tib-152, Molt-4, Hl-60, L- 1210 which were incubated in the presence of genistein (the reference substance) or tested substances in concentrations from 1 to 150 μM/1. Incubation time was 12 to 72 hours. Then the cells were stained with a set of fluorescent FDA stains (fiuorescein diacetate) and PI (propidium iodide). FDA stains alive cell, while PI - dead cells. (FDA/PI method is routinely used for cytotoxicity examination: Kenneth H., Senft JA., SenftJ.: J. Histochem. And Cytometry 33, 77, 1985). The stained samples were analysed by flow cytometry. The results were presented in two- dimensional cytograms and analysed taking into consideration the following criterions: a degree of FDA staining, a degree of PI staining, size and granularity of the cells. Each experimental point was repeated twice, and 5000 - 10000 cells were analysed during cytometric examination. Survival rate in the control culture was higher than 90%.
Results
The potency of toxic activity (relative cytotoxicity) of the tested compounds was expressed with regard to genistein, as a ratio of percentage of alive cells in a culture with genistein to percentage of alive cells in a culture with the tested compound.
The influence of the examined compounds on the rate of proliferation (relative cytostatic activity) was expressed with regard to genistein as a ratio of cell culture density in the presence of genistein to cell density in the presence of the tested compound.
The results of the experiments are summerized in Table 1.
Tablel
Figure imgf000009_0001
n.act.* - non-active compound
Antyproliferative (cytostatic) and antitumour (cytotoxic) activity of genistein derivatives exhibited in vitro unequivocally indicate the possibility of their potential use in clinical oncology (chemotherapy of neoplasms) and/or chemoprevention of neoplasms. Moreover, the invention includes the pharmaceutical preparation containing the active substance which is the genistein derivative of formula 1, wherein Ri and R2 are the same or different and independently represent:
- hydrogen atom
- alkyl - allyl
- aryl
- alkyloaryl
- alkylcarbonyl
- arylcarbonyl while each of the above mentioned groups may be substituted in a chain or ring by amino, nitro or nitrile groups;
- R5(R6)R7-Si- group, wherein R5, R6 and R7 are the same or different and denote alkyl C1-6 or aryl, - mono-, di- or oligosaccharide group, while at least one hydroxyl group of saccharide group may be substituted by the same or different acyl, alkyl, acyloxyalkyl or aryl groups; R3 denotes hydrogen atom or -COCH3 group, and Ri denotes hydrogen atom, - SO3H, SO3 ", -NH2 or -NO2 group or its pharmaceutically acceptable salt, together with the usual carriers and/or auxiliaries.
The preparation according to the invention may be in a pharmaceutical dosage form suitable for oral, parenteral, intranasal, sublingual or rectal use or for administration by inhalation or insufflation. Especially the preparation may be in the dosage form of a tablet, pill, capsule, powder, granules, sterile solution or suspension, aerosol, ampoule, liposomal preparation or suppository.
Appropriate pharmaceutical dosage forms of the preparation according to the invention are prepared by the methods known by those skilled in the art.
Solid dosage forms, such as tablets, pills, powders, granules or capsules are prepared by mixing the active substance with the pharmaceutically acceptable carrier, such as corn starch, lactose, saccharose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums and other pharmaceutical diluents, for example water, in order to obtain solid preliminary mixture containing homogenous mixture of the compound according to the invention or its pharmaceutically acceptable salt. The mixture obtained in this way may be then tabletted or drageetted or the capsules may be filled with it. Tablets or granules of the new composition may be coated or processed in another way in order to obtain unit form ensuring preferable prolonged action. A range of various substances may be used for preparation of such protective or coating layers including various polymeric acids and mixtures of polymeric acids with such substances as shellac, cetyl alcohol or cellulose acetate.
Liquid dosage forms appropriate for oral or parenteral administration of the preparations according to the invention include water solutions, syrups, water or oil suspensions, emulsions with edible oils such as cotton seeds oil, sesame oil, coconut or peanut oil as well as elixirs with similar pharmaceutical carriers. Appropriate dispergating or suspending agents for water suspensions include synthetic or natural gums such as tragacanth, acacia, alginate, dextran, carboxymethyl sodium cellulose, methylcellulose, polyvinyl pyrrolidone or gelatine.
Pharmaceutical preparations according to the invention may be used in chemotherapy and/or chemoprevention of neoplasms.
The invention is further illustrated by the following examples.
Example 1
7-tert-Butyldimethylsilyloxy-5-hydroxy-3-(4'-hydroxyphenyl)chromen-4-on (2a) and 5,7-dihydroxy-3-(4'-t-utyldimethylsilyloxyphenyl)chromen-4-on (2b)
Imidazole (1.36g, 20 mmol) and tert-butyldimethylsilyl chloride (1.65 g, 11 mmol) were added to genistein solution 1(2.7 g, 10 mmol) in anhydrous N,N- dimethylformamide (DMF), the reaction was carried out at room temperature for 24 hours. After that time the reaction solution was diluted by toluene (300 ml) and rinsed with water (2 x 50 ml). Toluene solution was dried with anhydrous MgSO4, concentrated on a rotary evaporator and the residue of 0.3 g of yellow oil soldifying at room temperature was purified by column chromatography method (flash) eluting with the mixture of hexa ethyl acetate (10:1 v/v). 0.27 g (yield: 7%) of the mixture of products 2a and 2b was obtained in the form of white crystals. For compounds 2a and 2b: 1H NMR (CDC13, δ (ppm); 0.21 (s, 6H, CH3), 0.99 (s, 9H, t-Bu), 6.33-638 (m, 2H, H-6, H-8), 6,86-6.92 (m, 2H, H-3', H-5'), 7.38 (d, 2H, H-2'. H-6', J=8.6 Hz), 7.85 (s, 1H, H-2), 12.92 (s.lH, 5-OH). HR MS for C21H24O5Si: calculated 384.1393, found 384.1382; M/z 384 (M+), 327 (100%).
Example 2 7-tert-Butyldimethylsilyloxy-5-hydroxy-3-(4'-t- butyldimethylsilyloxyphenyl)chromen-4-on (3) Following the procedure described in Example 1, imidazole (2.72 g, 40 mmol) and tert-butyldimethylsilyl chloride (3.3 g, 22 mmol) were added to genistein solution 1 (2.7 g, 10 mmol) in DMF (30ml). The reaction was carried out at room temperature for 1.5 hours. 5.34 g of yellow precipitate was obtained, it was purified by chromatography with eluting mixture of hexamethyl acetate (20:1 v/v). 4.19 g of the product 3 was obtained (yield:84%) and 0.26 g (yield:6%) of the mixture of products 2a and 2b was obtained in the form of white crystals. For compound 3: 1H NMR (CDC13), δ (ppm); 0.23 (s, 6H, CH3), 0.28 (s, 6H, CH3)0.99 (s,18H, t-Bu), 6.31 (d, IH, H-6, J=2.2 Hz), 6.35 (d, IH, h-8, J=2.2 Hz), 6.91 (d, 2H, H-3', H-5', J=8.6 Hz), 7.39 (d, 2H, H-2', H-6', J=8.6 Hz), 7.86 (s, IH, H-2), 12.82 (s, IH, 5-OH).
HR MS for C27H38O5Si2: calculated 498.2257, found 498.2248; m/z 498 (M+), 441 (100%).
Example 3
7-tert-Butyldimethylsilyloxy-5-acetoxy-3-(4'-acetoxyphenyl)chromen-4-on (4) 7-t-butyldimethylsilyloxy-5-hydroxy-3-(4'-acetoxyphenyl)chromen-4-on (5)
Acetic anhydride (1.5ml, 16 mmol) was added to the solution of compound 3 (4g, 8 mmol) (obtained according to the example 2) in anhydrous pyridine (15 ml). The reaction was carried out at room temperature for 3 hours. After that time the mixture was diluted with toluene (100 ml) and rinsed with cold water (3 x 30 ml). Toluene solution was dried with anhydrous MgSO4, concentrated under vacuum on the evaporator and additionally evaporated with toluene (2 x 10 ml). The obtained yellow solid (4.03 g) was purified on chromatographic column with use of eluting mixture of hexamethyl acetate (15:1 vol). 2.32g (yield: 62%) of product 4 in the form of white crystals and 1.02 g (yield: 30%) of product 5 in the form of light yellow crystals was obtained.
For compound 4: 1H NMR (CDC13), δ (ppm); 0.21 (s, 6H, CH3), 0.99 (s, 9H, t-Bu), 2.34 (s, 3H, CH3CO), 6.84 (d, IH, J=2.4 Hz), 6.87 (d, 2H, H-3'5 H-5', J=8.6 Hz), 7.23 (d, IH, h-6, J=2.4 Hz), 7.34(d, 2H, H-2', H-6', J=8.6 Hz), 7.85 (s, IH, H-2). HR MS for C25H28O7Si: calculated 468.1604, found 468.1620; M/z 468 (M+), 426 (54%), 369 (100%), 327 (99%). For compound 5: 1H NMR (CDC13), δ (ppm), 0.23 (s, 6H, CH3), 1.00 (s, 9H, tBu), 2.33 (s, 3H, CH3CO), 6.58 (d, IH, H-6, J=2.2 Hz), 6.75 (d, IH, H-8, J=2.2), 6.92 (d, 2H, H-3', H-5', J=8.6 Hz), 7.40 (d, 2H, H-2', H-6', J=8.6 Hz), 7.94 (s, IH, 5-OH). HR MS for C23H26O6Si: calculated 426.1498, found 426.1506; m/z 426 (M+), 369 5 (62%), 327 (100%).
Example 4 7-Hydroxy-5-acetoxy-3-(4'-acetoxyphenyl)chromen-4-on (6)
" 10 1M water solution of BU4NF (0.8 ml, 0.8 mmol) was added to the solution of compound 4 (0.8 g, 1.6 mmol) (obtained according to Example 3) in THF. The solution was stirred at room temperature while controlling the reaction course by TLC method. After several minutes the reaction was ended, the reaction mixture was diluted with methylene chloride (50 ml) and rinsed with water (20 ml).
15 Methylene chloride solution was dried with anhydrous MgSO4, concentrated and the residue in the form of precipitate was purified by column chromatography method using eluting mixture of benzene: ethyl acetate (4:1 /). 0.3 g of product 6 (yield: 53%) was obtained in the form of white crystals. For compound 6: 1H NMR (CDCI3), δ (ppm), 2.28 (s, 3H, CH3CO), 2.29 (s, 3H,
20 CH3CO0, 6.60 (d, IH, H-6, J=2.38 Hz), 6.80 (d, IH, H-8, J2.38 Hz), 7.21 (d, 2H, H-3', H-5', J=8.6 Hz), 7.50 (d, 2H, H-2', H-6', J=8.6 Hz), 8.36 (s, IH, H-2), 11.19 (s, IH, 7-OH).
HR MS for C19H14O7: calculated 354.0739, found 354.0722; m/z 354 (M+), 312 (37%), 27- (100%).
25
Example 5 7-O-Benzoil-4'-O-tert-butyldimethylsilylgenistein (7)
Following the procedure described in Example 3, anhydride of benzoic acid
30 (452 mg, 2 mmol) was added do the solution of 7,4'-di(O-tert- butyldimethylsilyl)genistein (3) in anhydrous pyridine. The solution was stirred at room temperature for 24 hours. The raw product 7 was purified by chromatography with use of eluting mixture of hexan: ethyl acetate. 312 mg (yield: 64%) of genistein monobenzoate was obtained. (7).
For compound 7: UN nm (log ε) 257 (4.62), 328; IR (KBr) 3073, 2955, 2929, 2858, 1742 and 1670 cm-1; MS (LS S(+) NBA); m/z 489 (M.+H)+;
Properties of the compounds obtained in examples 1-5 are presented in table 2. Rt for all compounds is hydrogen atom.
Figure imgf000014_0001
Example 6 7-O-Genistein palmitate
7,4'-O-Genistein dipalmitate
Following the procedure described in Example 3, palmitoyl chloride (328 mg,1.2 mmol) was added to gemstein solution (1) (270 mg, 1 mmol) in anhydrous pyridine (5 ml). The solution was stirred at room temperature for 24 hours. The raw mixture of 565 mg of mono- and diacyl derivative of genistein (8 and 9) was separated by chromatography with use of eluting mixture of chloroform: ethanol
(98:2 v/v). 125 mg of compound 8 (yield 24.6%) and 190 mg of compound 9
(yield:25.5%) was obtained in the form of white precipitates.
For compound 8: 1H NMR (CDCI3), δ (ppm), 0.9 (t, 3H, J=6.2 Hz), 1.16-1.36 (m, 24H), 1.7 (m, 2H), 2.58 (t,2H, J=7.4 Hz), 6.56 (d, IH, J=2.2 Hz), 6.7 (d, IH, J=2.2
Hz), 6.85 (dd, 2H, J^δ.8 Hz, J2=2.2 Hz), 7.4 (dd, 2H, Jr=8.8 Hz, J2=2.2 Hz), 7.9 (s,
IH), 12.8 (s, IH);
HR MS for C3ιH40O6: calculated 508.2825, found 508.2837;
M/z 508 (M+), 270 (100%). For compound 9: 1H NMR (CDC13), δ (ppm); 0.9 (t, 6H, J=6.2 Hz), 1.16-1.36 (m, 48H), 1.7 (m, 4H), 2.58 (t, 4H, J=7.4 Hz), 6.56 (d, IH, J=2.2 Hz), 6.7 (d, IH, J=2.2 Hz), 7.17 (dd, 2H, ^=8.8 Hz, J2=2.2 Hz), 7.54 (dd, 2H, Jι=8.8 Hz, J2=2.2 Hz), 7.96 (s, IH), 12.8 (s, IH); HR MS (LSIMS(+), [M.+HJ+: for C47H7ιO7: calculated 747.5199, found 747.5249; m z 747 [M.+H]+.
Example 7
Genistein 7-O-palmitate (8) Following the procedure described in Example 6, potassium tert-butanolate
(45 mg, 0.4 mmol) was added to the genistein solution (1) (108 mg, 0.4 mmol) in N,N-dimethylformamide (DMF) (15 ml) and the mixture was stirred at room temperature for 1 hour. Then palmityloyl chloride (121 mg, 0.44 mmol) was added to the mixture and stirring was continued at room temperature for 24 hours. After that time the mixture was poured on water with ice (75 g) and extracted with 2 x 30 ml of mixture of diethyl ether: ethyl acetate (1:1 v/v). Collected extracts were rinsed with saturated solution of NaHCO3, water, dried with anhydrous sodium sulfate, concentrated on the evaporator and the residue was purified by chromatography. 90 mg of 7-O-palmitate derivative of genistein (8) (yield: 44.3%) was obtained in the form of white precipitate for which spectioscopic 1H NMR data are consistent with those described for compound 8 obtained according to Example 6.
Example 8
Genistein 4'-O-palmitate (10) Following the procedure described in Example 7, 95 mg (yield: 46.8%) of
4'-O-palmitate derivative of genistein (10) was obtained in the form of white precipitate in the reaction of genistein (1) (108 mg, 0.4 mmol) with palmityloyl chloride (121 mg, 0.44 mmol) in the presence of potassium tert-butanolate (148 mg, 1.32 mol). For compound 10: 1H NMR (CDC13), δ (ppm), 0.9 (t, 3H, J=6.2 Hz), 1.26-1.35 (m, 24H), 1.7 (m, 2H), 2.58 (t, 2H, J=7.4 Hz), 6.24 (d,lH, J=2.0 Hz), 6.30 (d, IH, J=2.0 Hz), 7.15 (d, 2H, J=8.4 Hz), 7.5 (d, 2H, J=8.4 Hz), 7.8 (s, IH), 12.8 (s, IH); HR MS for C31H40O6: calculated 508.2825, found 508.2800;m/z 508 (M+), 270 (100%).
Example 9 Genistein 4 ' -O-anthranilate (11)
Following the procedure described in Example 7, isatin anhydride (0.897 g,
5.5 mmol) and catalytic amount of 4-dimethylaminopyridine (DMAP) (61 mg,0.5 mmol) were added to genistein solution (1) (1.35 g, 5 mmol). The mixture was stirred at 95-100°C for 6 hours. After cooling to room temperature, the mixture was poured on water with ice (90 g), raw precipitate of product 11 was filtered, rinsed with water, dried and purified by chromatography with use of eluting mixture of chloroform: ethanol (100:2 v/v). 1.33 g (yield:68%) of compound 11 was obtained in the form of light-yellow precipitate.
For compound 11: 1HNMR (DMSO-d6), δ (ppm); 6.25 (d, IH, J=2.2 Hz), 6.42 (d, IH, J=2.2), 6.6 (m., IH), 6.7 (bs, 2H), 6.84 (dd, IH, J=8.6 Hz), 7.3 (m., 3H), 7.6
(dd, 2H,
Figure imgf000016_0001
1.6 Hz), 8.49 (s, IH),
10.95 (bs, IH), 12.88 (s, IH);
UN run (log ε) 259 (4.57); TR (KBr) 3458, 3352, 1689, 1652 and 1620 cm"1;
HR MS for C22Hι5O6Ν: calculated 389.0899, found 389.0882; m/z 389 (M+).
Properties of the compounds obtained in Examples 6-9 are presented in Table 3. 4 is hydrogen atom in all compounds. Table 3
Figure imgf000016_0002
Example 10
7-O-Pivaloyloxymethylgenistein (12a) and 4'-O-pivaloyloxymethylgenistein (12b) Genistein solution (1) (270mg, 1 mmol) in DMF (3 ml) with addition of diisopropylethylamine (DIPEA) (155 mg, 1.2 mmol) was stirred at room temperature for 20 minutes. After that time, pivaloyloxymethyl chloride (180, 1.2 M) and catalytic amount of 4-dimethylaminopyridine (DMAP) were added to homogenous mixture. The mixture was stirred at room temperature for 5 days. Then the mixture was poured on water with ice (50 g) and agitated with methylene chloride (2x 25 ml). Combined extracts were rinsed with water to neutral reaction, dried with anhydrous Na2SO4 and concentrated on the evaporator. The oily residue was dissolved in methylene chloride and injected on a column filled with silica gel. Products of genistein alkylation (12a and 12b) were washed out with the mixture of chlorofornr.ethanol (100:2 v/v). After concentration of the fraction, 120 mg (yield: 31%) of white-cream precipitate of the mixture of compounds 12a and 12 b was obtained. For compounds 12a and 12b: UN nm (log ε) 260 (4.59); IR (KBr) 3438 2973 1733 lόSSand lό^ cm"1;
HR MS (LS S(+), [M.+H]+) : for C21H21O7: calculated 385.1287, found 385.1299; m/z 385 [M.+H]+.
Example 11
7-O-Allylgenistein (13)
Following the procedure described in Example 10, the mixture of genistein
(1) (270 mg, 1 mmol) with allyl bromide (145 mg, 1.2 mmol) in the presence of diisopropylethylamine (155 mg, 1.2mmol) in DMF (5 ml) was heated at 60°C for 24 hours. The raw product 13 was purified by chromatography with use of eluting mixture of hexa ethyl acetate (7:3 v/v). 150 mg (yield: 48%) of compound 13 was obtained in the form of white-cream precipitate.
For compound 13: 1H ΝMR (DMSO-d6), δ (ppm); 4.7 (d, 2H, J=5.2 Hz), 5.27-5.47 (m, 2H), 5.95-6.14 (m., IH), 6.43 (d, IH, J=2.2 Hz), 6.68 (d, IH, J=2.2 Hz), 6.81(d,
2H, J=8.6 Hz), 7.38 (d, 2H, J= 8.6 Hz), 8.41 (s, IH), 9.62 (bs,lH), 12.95 (s, IH);
UN nm (log ε) 262 (4.57); IR (KBr) 3126, 1666, 16121573 and 1517 cm-1. Example 12 7-O-AUyl-5-acetoxy-3-(4'-acetoxyphenyl)clιromen-4-on (14)
Acetate anhydride (5 ml) was added to the solution of compound (13) (310 mg, 1 mmol) in anhydrous pyridine (5 ml). The mixture was stirred at room temperature for 24 hours. Then the mixture was diluted with ethyl acetate (50 ml), rinsed with water (50 ml), 1M solution of hydrochloric acid, saturated solution of NaHCO3 and brine. The extract of ethyl acetate was dried with anhydrous Na2SO4, concentrated on the evaporator and the residue was purified by chromatography with use of eluting mixture of hexa ethyl acetate (7:3 v/v). 315 mg (yield: 80%) of compound 14 was obtained in the form of white precipitate.
For compound 14: 1H NMR (CDC13), δ (ppm); 2.31 (s, 3H), 2.41 (s, 3H), 4.63 (d, 2H, J=5.3 Hz) 5.33-5.49 (m, 2H), 5.95-6.15 (m, IH), 6.64 (d, IH, J=2.4 Hz), 6.79 (d, IH, J=2.4 Hz), 7.14 (d, 2H, J=8.6Hz), 7.83 (s,lH); UN nm (log ε) 251 (4.45); IR (KBr) 3123, 3081 1773, 1644 1564 and 1507 cm"1. MS (LSIMS(+), NBA): m/z 395 [M.+H]+.
Example 13 4'-O-[2-(3-Cyano-5-nitropyridyl)]genistein (15)
Following the procedure described in Example 10, the mixture of genistein (1) (540 mg, 2 mmol), 2-chloro-3-cyano-5-nitropyridine (403 mg, 2.2 mmol) in the presence of diisopropylethylamine (400 mg,3.1 mmol) in DMF (6 ml) was heated at 80°C for 3.5 hours. Then the mixture was chilled to room temperature and poured on water with ice (60g). The precipitate of product 15 was filtered, rinsed with water and dried. 718 mg (yield: 86%) of product 15 was obtained in the form of light-beige precipitate. The analytical sample (100 mg) was purified by chromatography with use of eluting mixture of hexan: acetate (2:1 v/v). For compound 15: 1H NMR (DMSO-d6), δ (ppm), 6.25 (d, IH, J=2.2 Hz), 6.43 (d, 1H,J= 2.2 Hz), 7.41 (d, 2H, J=8.7 Hz), 7.70 (d, 2H, J=8.7 Hz), 8.51 (s, IH), 9.25 (d, IH, J=2.7), 9.35 (d, IH, J=2.7), 9.95 (bs,lH), 12.85 (s, IH);
UN nm (log ε) 262 (4.57); IR (KBr), 3382, 3082, 2248 (CΝ), 1654, 16051578 and 1516 cm"1; MS (LSrMS(+), NBA): m/z 418 [M.+H]+.
Example 14
Caesium salt of genistein 4'-O-propylsulfonate (16)
The mixture of caesium genisteinate (100 mg, 0.25 mmol) (obtained in the form of cream precipitate in the reaction of genistein (270 mg, 1 mmol) with caesium carbonate (162 mg, 0.5 mmol) in methanol (10 ml) carried out at room temperature for Vz hour and evaporating methanol to dryness), propano-l,3-diyl sulfate (42 mg, 0.3 mmol) in DMF (3 ml) was heated at 60°C for 1.5 hours. Then the mixture was evaporated under vacuum and the residue was crystallised from ethanol. 85 mg (yield: 65%) of compound 16 was obtained in the form of crystalline precipitate.
For compound 16: 1H NMR (DMSO-d6), δ (ppm); 1.87 (t, 2H, J=6.0 Hz), 3.55 (t, 2H, J=6.0 Hz), 4.15 (t, 2H, J=6.0 Hz), 6.39 (d, IH, J=2.2 Hz), 6.64 (d, IH, J=2.2
Hz), 6.82 (d, 2H, j=8.6 Hz), 7.38 (d, 2H, J=8.6 Hz), 8.39 (s, IH), 9.59 (bs, IH),
12.95 (s, IH);
UN nm (log ε) 262 (4.38); TR (KBr), 1668, 1573 and 1520 cm"1;
Properties of the compounds obtained in Examples 10-14 are presented in
Table 3. 4 is hydrogen atom for all compounds.
Table 3
Figure imgf000019_0001
Figure imgf000020_0001
Example 15
3'-Nitrogenistein (17)
Solution of 65% nitric acid (1 ml) was added to the mixture of genistein (1)
(270 mg, 1 mmol) in glacial acetic acid (3 ml) and the mixture was stirred for 2 hours at 0-5°C, and then at room temperature for 4 hours. After that time the mixture was poured to water with ice (40 g). The precipitate of product 17 was filtered, rinsed with water and dried. 242 mg (yield: 77%) of compound 17 was obtained in the form of yellow precipitate. The analytical sample (100 mg) was purified by chromatography with use of the eluting mixture of hexan: acetate (2:1 v/v).
For compound 17: 1H NMR (DMSO-d6), δ (ppm), 6.07 (d, IH, J=2.0Hz), 6.25 (d, IH, J=2.2 Hz), 7.02 (d,lH, J=8.79 Hz), 7.58 (d, IH, Jl=8.6Hz, J2=2.2 Hz), 7.97 (d,
IH, J=2.2 Hz), 8.33 (s, IH), 10.8 (bs, IH), 11.05 (bs, IH), 12.6 (bs, IH);
UN nm (log ε) 262 (4.57); IR (KBr), 3515, 3449, 3279, 3087, 2656, 1663, 1633,
1612, 1581, 1537 and 1495 cm"1;
Example 15 3'-Sulfogenistein (18)
Genistein suspension (1) (270 mg, 1 mmol) in concentrated sulfuric acid (5 ml) was stirred at room temperature for 12 hours. After that time the mixture was poured to water with ice (50 g) and extracted with the mixture of n-butanol: ethyl acetate (8:2 v/v)2 x 25 ml. The combined extracts were rinsed with the mixture of brine:water (6:4 v/v) 3 x 25 ml to pH = 2.5 and dried with anhydrous sodium sulfate. After removal of solvents to dryness, 213 mg (yield: 60%) of compound 18 was obtained in the form of crystalline precipitate. For compound 18: 1H NMR (DMSO-d6), δ (ppm); 6.22 (d, IH, J=2.2 Hz), 6.34 (d, IH, J=2.2 Hz), 6.81 (d, IH, J=8.4 Hz), 7.37 (dd, IH, Jl=8.4 Hz, J2=2.3 Hz), 7.67 (d, IH, J=2.2 Hz), 8.4 (s, IH), 10.6 (bs, IH), 10.85 (s, IH), 12.95 (s, IH); UN nm (log s) 262 (4.49); IR (KBr), 1665, 1633, 1612, 1512, 1437 and 1197 cm"1.
Properties of the compounds obtained in Examples 15-16 are presented in
Table 5.
Table 5
Figure imgf000021_0001
Example 17
7-O-(3 ",4"-di-O-Acetyl-6-deoxy-glucopyranosyl)genistein (19)
3,4-di-O-Acetyl-6-deoxy-L-glucal (428 mg, 2 mmol) and catalytic amount of triphenylphospbine hydrobromide (PPri3xHBr) (34 mg, 0.1 mmol) were added to genistein solution (1) (270 mg, 1 mmol) in anhydrous THF (5 ml). The mixture was stirred at 50°C for 24 hours. After cooling to room temperature, the mixture was poured water with ice (50 g) with addition of saturated solution of NaHCO3, the neutral mixture was rinsed with ethyl acetate (50 ml) and water. The extract of ethyl acetate was dried with anhydrous Na SO4, concentrated and the residue was purified by chromatography with use of the eluting mixture of chloroform:ethanol (100:2 v/v). 90 mg (yield: 18%) of compound 19 was obtained in the form of colourless precipitate. For compound 19: 1H NMR (CDC13), δ (ppm), 1.16 (m, 3H), 2.07 (s, 6H), 2.46 (m, IH), 3.94 (m, IH), 4.84 (t, IH), 5.47 (m, IH), 5.62 (d,lH), 6.36 (d, IH, J=2.2 Hz),
6.39 (d, IH, J=2.2 Hz), 7.12 (d, 2H, J=8.6 Hz), 7.42 (d, 2H,J=8.6 Hz), 7.79 (s, IH),
9.40 (bs, IH), 12.78 (s, IH);
UN nm (log ε) 261 (4.53); IR (KBr) 1745,1654, 1512 and 1369 cm"1. Example 18
7-O-(2' ' ,3 " ,5 " -tri-O-Benzyl-arabinofuranosyl)genistein (20)
Catalytic amount of SnCl4 (333mg,1.28 mmol) was added to the mixture of genistein (1) (270 mg, 1 mmol), 2,3,5-tri-O-benzylarabinofuranose(428 mg, 2 mmol) in anhydrous acetonitrile (5 ml) cooled to °C. Then the mixture was stirred at room temperature for 40 minutes. After that time the mixture was poured on water with ice (30 g) with addition of saturated solution of NaHCO3 (20 ml), the neutral mixture was rinsed with ethyl acetate (50 ml) and water. The extract of ethyl acetate was dried with anhydrous Na2SO , concentrated and the residue was purified by chromatography with use of eluting mixture of hexa ethyl acetate (8:2 v/v). After evaporation of appropriate fractions, 150 mg (yield: 22%) of compound 20 was obtained in the form of colourless precipitate. For compound 20: 1H NMR (CDC13), δ (ppm); 3.69 (d, 2H), 3.99 (, IH), 4.19-4.28 (m, 2H) 4.26 (q, 2H), 4.48 (q, 2H), 5.59 (s, 2H), 6.41 (s, IH), 6.89 (d, 2H), 7.05 (m, 2H), 7.18-7.35 (m, 14H), 7.39 (d, 2H), 7.83 (s, IH), 9.79 (s, IH), 13.21 (s, IH); HR MS (LSIMS(+), {M. +H]+): for C41H37O9: calculated 673.2437, found 673.2452; m z 673 [M.+H]+.
Example 19
4'-O-(2",3 ",5"-tri-O-Acetyl-α,β-D-riboruranosil-genistein and
4'-O-(2",3",5"-tri-O-Acetyl-α,β-D-ribofuranosil)genistein (21a, 21b)
4.36 ml of 0.87M solution of SnCl4 in CH2C12 (990 mg, 3.8 mmol) was added at room temperature to anhydrous solution of tetraacetylribose (954.8mg, 3.0 mmol) in 40 ml of methylene chloride. After 20 minutes this solution was added to stirred genistein suspension (540.5 mg, 2.0 mmol) in anhydrous acetonitrile and clear, yellow solution was obtained. After 50 minutes of stirring at room temperature the reaction was ended by adding 40 ml of cold saturated solution of NaHCO3 and 80 ml of chloroform. The organic layer was separated, the water layer was rinsed with the mixture of chloroform-acetonitrile 4:1 (50 ml). The combined organic layers were dried over Na SO and then evaporated to dryness. The residue after evaporation was treated with chloroform and the obtained suspension was filtered. The filtrate was evaporated to oil which was dissolved in 5 ml of the mixture chloroform:ethanol 4:1 (v/v) and separated on chromatographic column with silica gel 60H (5 x 8 cm) in chloroform-ethanol gradient (from 98:2 to 9:1). After evaporation of the solvent, 550 mg (52%) of the main product was obtained, which was the mixture of α and β anomers in a ratio of 1 :2 (NMR).
Example 20 4'-O-(α-D-Ribofuranosil)genistein and
4'-O-(β-D-Ribofuranosil)genistein (22a i22b).
The raw mixture of α and β anomers of triacetylribogenistein obtained in the previous example (400 mg, 0.757 mmol) was dissolved in 12 ml of methanol and 4 ml of 25%) NH OH was added. After 24 hours at room temperature, a complete unblocking of hydroxyl groups was determined on the base of TLC. The mixture was evaporated to dryness, then it was co-evaporated with water (2 x 50 ml) and with ethanol (2 x 50 ml). The obtained oil was dissolved in ethanol (100 ml) and adsorbed on silica gel (70-230 mesh) by evaporation. The product was separated on a column with silica gel 60H (3.5 xl5 cm) in toluene-ethanol system (4: 1). The following products were obtained:
128.8 mg (42%) of anomer β in the form of white precipitate which was crystallised from water, then fine needles of slightly pink colour were obtained, t.t. 199.5- 201°C, purity according to HPLC- 98.9%. 1H-NMR (DMSO-d6), δ (ppm): 12.91 (s, IH); 10.92(s, IH), 8.38 (s, IH), 7.49 (d, 2H), 7.04 (d, 2H), 6.40 (d, IH); 6.23 (d, IH), 5.51 (s, IH); 5.36 (d, IH), 5.03 (d, IH), 4.70 (t,lH); 4.02 (m, 2H); 3.91 (m, IH); 3.54 (m, IH); 3.36 (m, IH); 64.4 mg (21.1% ) of anomer α which was crystallised from 30% ethanol to obtain very fine pink needles of t.t. 158-160°C, purity according to HPLC - 90.0%. 1H-NMR (DMSO-d6), δ (ppm): 12.92 (s, IH); 10.93 (bs, IH); 8.39 (s, IH); 7.49 (d, 2H); 7.11 (d, 2H); 6.40 (d, IH); 6.24 (d, IH); 5.63 (d, IH); 4.72 (d, IH); 4.92 (d, IH); 4.82 (t, IH); 4.07 (m, IH), 3.96 (m, 2H), 3.49 (m, 2H). Properties of the compounds obtained in Examples 17-20 are presented in Table 6. 4 is hydrogen atom in all compounds.
Table 6
Figure imgf000024_0003
Structures of substituents in formulae of compounds 19-22 are presented below.
19: R1= a:
Figure imgf000024_0001
20: Rχ= b:
21a: R2= C:
21b: R2= d:
Figure imgf000024_0002
b: R2= e:
Figure imgf000025_0001
b: R2= f
Figure imgf000025_0002

Claims

Claims
1. Genistein derivatives of formula 1 wherein R and R2 are the same or different and independently represent:
- hydrogen atom,
- alkyl,
- aryl,
- alkyloaryl, - alkylcarbonyl,
- arylcarbonyl, while each of the above mentioned groups may be substituted in a chain or ring by amino, nitro or nitrile groups,
- R5(R6)R7-Si- group wherein R5, R6 and R7 are the same or different and denote C1-6alkyl or aryl,
- mono-, di- or oligosacccharide group while at least one hydroxyl group of saccharide group may be substituted by the same or different acyl, alkyl, acyloxyalkyl or aryl groups;
R3 represents hydrogen atom or -COCH3 group; and Rt represents hydrogen atom, -SO3H, SO3 " or -NH2 or -NO group; and their pharmaceutically acceptable salts.
2. Genistein derivatives of formula 1 according to claim 1 wherein Ri represents alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl, R5(R6)R -Si- group, R2 represents hydrogen atom or -COCH3 group, and R3 and Rt are simultaneously hydrogen atom.
3. Genistein derivatives of formula 1 according to claim 1 wherein
Figure imgf000026_0001
and Rt are hydrogen atom, R3 is hydrogen atom or -COCH3 group, and R2 represents alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbonyl or R5(R6)R7-Si-.
4. Genistein derivatives of formula 1 according to claim 1 wherein Ri and R2 are the same or different and represent alkyl, allyl, aryl, alkyloaryl, alkylcarbonyl, arylcarbolnyl or R5(R6)R7-Si-, and R3 is hydrogen atom or -COCH3, and t is hydrogen atom.
5. Genistein derivatives of formula 1 according to claim 1 wherein Ri, R3 and Rt are hydrogen atom, and R2 is saccharide group.
6. Genistein derivatives of formula 1 according to claim 5 wherein R1? R3 and Rt are hydrogen atom, and R is furanose group.
7. Pharmaceutical preparation containing a genistein derivative of formula 1 wherein: Rt and R2 are the same or different and independently represent:
- hydrogen atom,
- alkyl,
- allyl, alkyloaryl, - alkylcarbonyl,
- arylcarbonyl, while each of the above mentioned groups may be substituted in a chain or ring by amino, nitro or nitrile groups,
- R5(R6)R7-Si- group wherein R5, R6 and R7 are the same or different and represent C1-6alkyl or aryl,
- mono-, di- or oligosaccharide group while at least one hydroxyl group of saccharide group may be substituted by the same or different acyl, alkyl, acyloxyalkyl or aryl groups;
R3 represents hydrogen atom or -COCH3, and R4 represents hydrogen atom, -SO3H, SO3 ", -NH2 or -NO2 groups or its pharmaceutically acceptable salt together with the usual carriers and/or auxiliaries.
PCT/PL2002/000029 2001-04-09 2002-04-08 New genistein derivatives and pharmaceutical preparations containing them WO2002081491A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PL346955A PL205635B1 (en) 2001-04-09 2001-04-09 Novel derivatives of genistein and pharmacological agents containing such derivatives
PLP-346955 2001-04-09

Publications (2)

Publication Number Publication Date
WO2002081491A2 true WO2002081491A2 (en) 2002-10-17
WO2002081491A3 WO2002081491A3 (en) 2004-02-26

Family

ID=20078583

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/PL2002/000029 WO2002081491A2 (en) 2001-04-09 2002-04-08 New genistein derivatives and pharmaceutical preparations containing them

Country Status (2)

Country Link
PL (1) PL205635B1 (en)
WO (1) WO2002081491A2 (en)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007515389A (en) * 2003-05-15 2007-06-14 成都迪康▲ヤオ▼物研究所 Isoflavone derivatives of tectorigenin, their preparation, and antiviral agents containing them as active ingredients
EP2373326A1 (en) * 2008-12-11 2011-10-12 Axcentua Pharmaceutucals AB Crystalline forms of genistein
US8476313B2 (en) * 2004-08-26 2013-07-02 The Ohio State University Research Foundation Heteroaryl-containing isoflavones as aromatase inhibitors
CN107652261A (en) * 2017-09-30 2018-02-02 桂林医学院 A kind of calycosin derivative and its synthetic method
WO2023212130A1 (en) * 2022-04-28 2023-11-02 GreenStone Biosciences, Inc. Treatment of the adverse cardiovascular effects of cannabinoids

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PL399467A1 (en) 2012-06-08 2013-12-09 3G Therapeutics Inc. The use of genistein for lowering the storage organic compounds in cells in the treatment and/or prevention of lysosomal storage diseases
CN104974204B (en) * 2015-06-19 2017-12-19 武汉光谷人福生物医药有限公司 The method and system of separating flavone oxygen glycosides compound monomer is extracted from Desmodium styracifolium

Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997029779A2 (en) * 1996-02-16 1997-08-21 Regents Of The University Of Minnesota Egf-genistein conjugates for the treatment of cancer
WO1997032872A1 (en) * 1996-03-05 1997-09-12 Imperial College Of Science Technology And Medicine Compounds with a sulfamate group
WO1998011876A1 (en) * 1996-09-18 1998-03-26 Marigen S.A. Bioflavonol glycoside peroxy esters and their preparation for pharmacologically effective concentrations and ultramicroemulsions
WO1998048790A1 (en) * 1997-04-28 1998-11-05 Anticancer, Inc. Use of genistein and related compounds to treat certain sex hormone related conditions
WO1999022728A1 (en) * 1997-10-31 1999-05-14 Arch Development Corporation Methods and compositions for regulation of 5-alpha reductase activity
FR2781154A1 (en) * 1998-07-15 2000-01-21 Lafon Labor Compositions containing isoflavonoids or chromones to prevent proliferation of clonogenic cells in tumors, as supplement to conventional chemotherapeutic treatment
WO2000066576A1 (en) * 1999-04-30 2000-11-09 G.J. Consultants Pty Ltd Isoflavone metabolites
WO2001017985A1 (en) * 1999-09-03 2001-03-15 Indena S.P.A. Novel derivatives of flavones, xanthones and coumarins
WO2001049281A2 (en) * 1999-12-30 2001-07-12 Proteotech, Inc. POLYHYDROXYLATED AROMATIC COMPOUNDS FOR THE TREATMENT OF AMYLOIDOSIS AND α-SYNUCLEIN FIBRIL DISEASES

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997029779A2 (en) * 1996-02-16 1997-08-21 Regents Of The University Of Minnesota Egf-genistein conjugates for the treatment of cancer
WO1997032872A1 (en) * 1996-03-05 1997-09-12 Imperial College Of Science Technology And Medicine Compounds with a sulfamate group
WO1998011876A1 (en) * 1996-09-18 1998-03-26 Marigen S.A. Bioflavonol glycoside peroxy esters and their preparation for pharmacologically effective concentrations and ultramicroemulsions
WO1998048790A1 (en) * 1997-04-28 1998-11-05 Anticancer, Inc. Use of genistein and related compounds to treat certain sex hormone related conditions
WO1999022728A1 (en) * 1997-10-31 1999-05-14 Arch Development Corporation Methods and compositions for regulation of 5-alpha reductase activity
FR2781154A1 (en) * 1998-07-15 2000-01-21 Lafon Labor Compositions containing isoflavonoids or chromones to prevent proliferation of clonogenic cells in tumors, as supplement to conventional chemotherapeutic treatment
WO2000066576A1 (en) * 1999-04-30 2000-11-09 G.J. Consultants Pty Ltd Isoflavone metabolites
WO2001017985A1 (en) * 1999-09-03 2001-03-15 Indena S.P.A. Novel derivatives of flavones, xanthones and coumarins
WO2001049281A2 (en) * 1999-12-30 2001-07-12 Proteotech, Inc. POLYHYDROXYLATED AROMATIC COMPOUNDS FOR THE TREATMENT OF AMYLOIDOSIS AND α-SYNUCLEIN FIBRIL DISEASES

Non-Patent Citations (22)

* Cited by examiner, † Cited by third party
Title
A.C.JAIN ET AL.: "A Study of Nuclear Prenylation and Allylation of Isoflavones and Synthesis of 4'-Methyl Ethers of Asajin and Warangalone." TETRAHEDRON, vol. 26, no. 8, 1970, pages 1977-1988, XP001109153 *
A.C.JAIN ET AL.: "Synthesis of Alpium Isoflavone, Osajin, and Warangalone." JOURNAL OF ORGANIC CHEMISTRY., vol. 39, no. 15, 1974, pages 2215-2217, XP001126263 AMERICAN CHEMICAL SOCIETY. EASTON., US ISSN: 0022-3263 *
B.S.BAJWA ET AL.: "Synthesis of Eupatoriochromene, Encecalin, Isoevodionol, Alloptaeroxylin & Related Compounds." INDIAN JOURNAL OF CHEMISTRY, vol. 9, 1971, pages 17-19, XP001126915 *
C.L.HOLDER ET AL.: "Quantification of Soy Isoflavones, Genistein and Daidzein, and Conjugates in Rat Blood Using LC/EC-MS." JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, vol. 47, no. 9, 1999, pages 3764-3770, XP001126265 *
CHEMICAL ABSTRACTS, vol. 132, no. 20, 15 May 2000 (2000-05-15) Columbus, Ohio, US; abstract no. 264129, "Production, isolation and structure elucidation of novel isoflavonoid compound K3-D4, K3-D5, K3-D6" XP002222414 & DATABASE CHEMICAL ABSTRACTS [Online] Chemical Abstracts Service, Columbus; C.A.132:264129 compound with RN 263554-80-9, & ZHONGGUO KANGSHENGSU ZAZHI, vol. 24, no. 4, 1999, pages 254-257, *
CHEMICAL ABSTRACTS, vol. 57, no. 1, 7 January 1963 (1963-01-07) Columbus, Ohio, US; abstract no. 9798h, A.VIGIER ET AL.: "Hydrolsyis of delta-oxonitriles and hydrogenation of delta oxoacids." XP002222415 & DATABASE CHEMICAL ABSTRACTS [Online] Chemical Abstracts Service, Ohio; & COMPT. REND., vol. 254, 1962, pages 4473-5, *
D.R.DOERGE ET AL.: "Analysis of Soy Isoflavone Conjugation in Vitro and in Human Blood using Liquid Chromatography-Mass Spectrometry." DRUG METABOLISM AND DISPOSITION, vol. 28, no. 3, 2000, pages 298-307, XP001126264 *
J.BORYSKI ET AL.: "A Regioselective Synthesis of Genistein 4'-O-Ribofuranosides." SYNTHESIS., no. 14, November 2001 (2001-11), pages 2170-2174, XP002222413 GEORG THIEME VERLAG. STUTTGART., DE ISSN: 0039-7881 *
J.YUN ET AL. : "Differential Inhibitors Effects of Sophoricoside Analogs on Bioactivity of Several Cytokines." LIFE SCIENCES, vol. 67, 2000, pages 2855-2863, XP002222411 *
K.NISHIYAMA ET AL.: "Syntheses of Isoflavones and Isoflavone Glycosides, and Their Inhibitory Activity Against Bovine Liver B-Galactosidase." BIOSCIENCE, BIOTECHNOLOGY AND BIOCHEMISTRY, vol. 57, no. 1, 1993, pages 107-114, XP001109133 *
L.FARKAS ET AL.: "About the Existence of Padmakastein and Padmakastin" TETRAHEDRON, vol. 25, no. 5, 1969, pages 1013-1019, XP001109134 *
L.FARKAS ET AL.: "Transacylation reactions in the flavonoid series. IV. New syntheses of 5-methylgenistein, prunetin, biochanin-A and sissotrin" ACTA CHIMICA ACADEMIAE SCIENTIARUM HUNGARICAE, vol. 60, no. 3, 1969, pages 293-299, XP001126906 *
LEWIS P ET AL: "SYNTHESIS OF ANTIOXIDANT ISOFLAVONE FATTY ACID ESTERS" SPECIAL PUBLICATION - ROYAL SOCIETY OF CHEMISTRY, ROYAL SOCIETY OF CHEMISTRY, LONDON, GB, vol. 240, 1999, pages 215-217, XP001122875 ISSN: 0260-6291 *
M.BRUM-BOUSQUET ET AL.: "Isolement et Etude du Sarothamnoside, Nouvel Heteroside d'Isoflavone de Divers Sarothamnus." PLANTA MEDICA, vol. 43, no. 4, 1981, pages 367-374, XP001126912 *
P.BEDNAREK ET AL.: "Profiling Changes in Metabolism of Isoflavonoids and their Conjugates in Lupinius albus Treated with Biotic Elicitor." PHYTOCHEMISTRY, vol. 56, no. 1, 2001, pages 77-85, XP001126919 *
P.T.LEWIS ET AL.: "Synthesis of Antioxidant Isoflavone Fatty Acid Esters" TETRAHEDRON, vol. 56, no. 39, 2000, pages 7805-7810, XP002222410 *
Q-H.MENG ET AL.: "Incorporation of Esterified Soybean Isoflavones with Antioxidant Activity into Low density Lipoprotein." BIOCHIMICA ET BIOPHYSICA ACTA, vol. 1438, 1999, pages 369-376, XP002222409 *
R.J.BASS ET AL.: "5-Acetoxyisoflavones and their Chloro-Derivatives." JOURNAL OF THE CHEMICAL SOCIETY. CHEMICAL COMMUNICATIONS., no. 6, 1979, page 264 XP002222412 CHEMICAL SOCIETY. LETCHWORTH., GB ISSN: 0368-1769 *
S-J. LEE ET AL.: "Antiinflammatory Activity of Isoflavanoids from Peuraria Radix and Biochanin A Derivatives." ARCHIVES OF PHARAMACAL RESEARCH , vol. 17, no. 1, 1994, pages 31-35, XP000886583 *
S-Y.NAMGOONG ET AL.: "Effects of Isoflavanoidson Mouse Lymphocyte Proliferation In Vitro." ARCHIVES OF PHARMACAL RESEARCH, vol. 17, no. 4, 1994, pages 236-239, XP001126903 *
T.TAKEDA ET AL.: "New Isoflavone Glycosides from the Woods of Sophora Japonica" PHYTOCHEMISTRY, vol. 16, no. 5, 1977, pages 619-620, XP001126918 *
Y.SHIBUYA ET AL.: "New Isoflavone Glucosides from White Lupin (Lupinus albus L.)" ZEITSCHRIFT F]R NATURFORSCHUNG, C: JOURNAL OF BIOSCIENCES, vol. 46, no. 7-8, 1991, pages 513-518, XP001122873 *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2007515389A (en) * 2003-05-15 2007-06-14 成都迪康▲ヤオ▼物研究所 Isoflavone derivatives of tectorigenin, their preparation, and antiviral agents containing them as active ingredients
US8476313B2 (en) * 2004-08-26 2013-07-02 The Ohio State University Research Foundation Heteroaryl-containing isoflavones as aromatase inhibitors
CN104860913A (en) * 2008-12-11 2015-08-26 艾克赛特药品有限公司 Crystalline forms of genistein
JP2015193630A (en) * 2008-12-11 2015-11-05 アクセンチュア ファーマシューティカルズ アーベー Crystalline forms of genistein
EP2373326A4 (en) * 2008-12-11 2012-09-05 Axcentua Pharmaceutucals Ab Crystalline forms of genistein
CN102292097A (en) * 2008-12-11 2011-12-21 艾克赛特药品有限公司 Crystalline forms of genistein
US9012495B2 (en) 2008-12-11 2015-04-21 Axcentua Pharmaceuticals Ab Crystalline forms of genistein
CN102292097B (en) * 2008-12-11 2015-05-27 艾克赛特药品有限公司 Crystalline forms of genistein
EP2373326A1 (en) * 2008-12-11 2011-10-12 Axcentua Pharmaceutucals AB Crystalline forms of genistein
JP2012512164A (en) * 2008-12-11 2012-05-31 アクセンチュア ファーマシューティカルズ アーベー Crystalline form of genistein
AU2009324489B2 (en) * 2008-12-11 2015-11-12 Axcentua Pharmaceuticals Ab Crystalline forms of genistein
US9492425B2 (en) 2008-12-11 2016-11-15 Axcentua Pharmaceuticals Ab Crystalline forms of genistein
CN104860913B (en) * 2008-12-11 2018-05-04 艾克赛特药品有限公司 The crystal form of genistein
CN107652261A (en) * 2017-09-30 2018-02-02 桂林医学院 A kind of calycosin derivative and its synthetic method
CN107652261B (en) * 2017-09-30 2020-08-07 桂林医学院 Calycosin derivative and synthetic method thereof
WO2023212130A1 (en) * 2022-04-28 2023-11-02 GreenStone Biosciences, Inc. Treatment of the adverse cardiovascular effects of cannabinoids

Also Published As

Publication number Publication date
PL346955A1 (en) 2002-10-21
WO2002081491A3 (en) 2004-02-26
PL205635B1 (en) 2010-05-31

Similar Documents

Publication Publication Date Title
EP2646456B1 (en) Process for the preparation of morpholinyl anthracycline derivatives
KR100252451B1 (en) A process for anomerizing nucleosides
US5036055A (en) Acylated derivatives of etoposide
EP0014853B1 (en) Anthracycline derivatives, a process for their preparation and pharmaceutical compositions containing them
JPS61204193A (en) Production of cytosine nuceoside
WO2002081491A2 (en) New genistein derivatives and pharmaceutical preparations containing them
WO1996005211A1 (en) Ganglioside gm3 analog having sialic acid residue fluorinated at the 9-position and intermediate therefor
US4351937A (en) N-Glycosyl derivatives of anthracycline antibiotics and the method of their preparation
Scheigetz et al. A Synthesis of 4-α-Guanidino-2-Deoxy-2, 3-Didehydro N-Acetylneuraminic Acid
KR900006217B1 (en) Nitrogen containing derivatives of epipododhyllotoxin glycodides
CA1306250C (en) 3',4'-dinitrogen substituted epipodophyllotoxin glucoside derivatives
Komiotis et al. Biologically Important Nucleosides: A General Method for the Synthesis of Unsaturated Ketonucleosides of Uracil and its Analogs
EP0350292B1 (en) Process for preparing 2'-deoxy-beta-adenosine
JPH0592987A (en) 4'-demethylepipodophyllotoxin glycosides
NZ227286A (en) Epipodophyllotoxin glucoside 4'-acyl derivatives and anti-tumour medicaments
Srivastava et al. Synthesis and biological evaluation of certain 2'-deoxy-. beta.-D-ribo-and 2. beta.-D-arabinofuranosyl nucleosides of purine-6-carboxamide and 4, 8-diaminopyrimido [5, 4-d] pyrimidine
JPH0692986A (en) Substituted nucleoside derivative, its production and medicinal composition containing it
US4522815A (en) Anthracycline glycosides
US5034380A (en) Alkoxymethylidene epipodophyllotoxin glucosides
Franzyk et al. Synthesis of Carbocyclic Homo‐N‐Nucleosides from Iridoids
Jiang et al. Synthesis and stereochemical assignments for 2-(α-and β-D-arabinofuranosyl) thiazole-4-carboxamides
FR2699535A1 (en) Etoposide derivatives, process for their preparation, their use as a medicament and their use for the preparation of a medicament for anticancer treatment
KR920001690B1 (en) Nitrosoures derivatives process for their preparation and medicaments containing them
EP2128170B1 (en) "Process for the glycosidation of colchicine and thiocolchicine"
CN108822170B (en) Anthraquinone benzimidazole nucleoside analogues and synthesis method and application thereof

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A2

Designated state(s): BA BG BR CA CZ EE GE HR HU IL IN JP KR LT LV MK MX NO RO SI SK UA US YU

AL Designated countries for regional patents

Kind code of ref document: A2

Designated state(s): AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase in:

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP