JPH10511346A - Chalcone and esters thereof with antiproliferative activity in uterus, ovary and breast tumors - Google Patents

Abstract

  (57) [Summary] The present invention relates to a number of natural or synthetic chalcones and saturated and unsaturated linear or branched fatty acids containing up to 22 carbon atoms in the treatment and prevention of uterine, ovarian and breast tumors. It relates to the use of esters, as well as the formulations containing them. Chalcones such as isocordoin, cordoin, 2-hydroxyderricin, 2 ', 4'-dihydroxychalcone, 4,2', 4'-trihydroxychalcone have marked affinity for the type II estrogen receptor as well as uterus, ovary , And has antiproliferative activity against breast tumor cell lines. These molecules have been found to be useful in both treating and preventing such tumors.

Description

The present invention relates to chalcone compounds or up to 22 carbons in the treatment and prevention of uterine, ovarian and breast tumors. The present invention relates to the use of saturated and unsaturated straight-chain or branched fatty acids containing atoms and their esters, and to formulations containing them. Recently, some flavonoids have been shown to have anticancer activity (Cancer Research 48, 5754, 1988) and chemopreventive activity (J. Nat. Prod. 53, 23, 1990) in some tumors. . In particular, quercetin, an almost ubiquitous flavonoid in plants, has been shown to enhance human leukemia cell proliferation (Br. J. of Haematology 75, 489, 1990) and other synergistic activities with conventional chemotherapy. Cell lines (Br. J. Cancer 62, 942, 1990-Int. J. Cancer 46, 1112, 1990-Cancer Chemother. Pharma macol. 28, 255, 1991-Gynecological Oncology 45, 13, 1991, 1992) showed some inhibitory activity. Although the mechanism of such inhibitory effects on proliferation is unknown, it appears to be involved in the interaction of this flavonoid with the type II estrogen receptor (J. Steroid Biochem. 30, 71, 1988). These sites were originally described by Clark in the rat uterus in 1978 (J. Biol. Chem. 253, 7630, 1978) and show the same steroid and tissue specificity, but they are estrogen receptor (ER) is present at higher concentrations and has a distinct affinity dissociation constant (KD: 10-20 nM) for estradiol that is lower than the ER (KD: 0.2-1 nM); different. This time, surprisingly, chalcone structural compounds, isocordoin, 4-hydroxyderricin, 2-hydroxyderricin, 3-hydroxyderricin, 2 ', 4'-dihydroxychalcone, 4,2', 4'-trihydroxychalcone And cordoin were found to have a markedly higher affinity for the type II estrogen receptor than the known products, with marked antiproliferative activity against uterine, ovarian, and breast tumor cell lines. The structures of the compounds listed above are as follows: Other chalcones strictly related to the former as far as their chemical structures are concerned, such as 4-hydroxycordoin and dihydrocordoin, whose structures are shown below, show no affinity for the above receptors. Accordingly, the present invention relates in a first aspect to the use of a compound of formula (I) for the preparation of a medicament having anticancer activity, in particular for the treatment of a tumor expressing the type II estrogen receptor. Provide use. The invention also provides, according to a further aspect, an ester of a compound of formula (I) with a saturated or unsaturated straight or branched chain fatty acid containing up to 22 carbon atoms. Particularly preferred are esters with acetic acid, butyric acid, palmitic acid or xymenic acid. The esters of the invention can be administered orally, and they appear to act as prodrugs of chalcone I. Finally, the invention provides a pharmaceutical composition comprising Compound I or an ester thereof as an active ingredient in admixture with suitable excipients. Compound (I) may be prepared by conventional methods as reported by Il Pharmaco, 30, 449-55, 1975; Il Pharmaco, 32, 261-69, 1977; Il Pharmaco, 30, 326-42, 1975. Can be. The affinity of Compound I for the type II estrogen receptor and its antiproliferative activity on ovarian tumor cells are reported in the table below. Evaluation of estrogen receptor binding has been performed on tumor cells of the ovary and other organs. Cells were grown in monolayer culture on minimal essential medium supplemented with 200 units / ml penicillin to keep the calf serum and medium sterile. To be able to replicate the test, these cells were treated with trypsin weekly, placed on plates at a density of 8 × 10 ⁻⁴ and cultured at 37 ° C. in an atmosphere containing 5% CO ₂ and moisture. . To assess the activity of the compounds, these cells were placed in wells (Falcon 3046, Becton Dickin son NY) at a concentration of 1 × 10 ⁻⁵ / ml with a minimal amount of substrate. After 24 hours, the substrate was replaced with fresh substrate and chalcone dissolved in absolute ethanol was added. Controls were similarly treated with the carrier without the active ingredient being tested. The above procedure was repeated at 24-hour time intervals during the 72-hour test period. After 24 hours, cells were incubated at 4 ° C. for 2.5 hours at 4 ° C. in the presence of only graded amounts of labeled estradiol ( ³ H-E2 40 Ci / mmol Amershan UK) or 100 volumes of diethylstilbestrol. At the end of the incubation period, the cells were quickly washed with fresh substrate and incubated with 1 M NaOH for 30 minutes. Radioactivity was measured by a scintillator and binding specificity was calculated as the difference between preparations with and without diethylstilbestrol. Results are expressed as number of binding sites per cell according to conventional methods. Inhibition on cell growth is assessed by direct calculation of cells comparing control growth to that of treated ones. Compounds of the invention inhibited cell proliferation in vivo, as shown by measuring the size of tumors implanted in nude athymic mice according to conventional conditions in the literature. Treatment of these animals at doses from 1 to 100 mg / kg clearly showed these significant regressions until the examined tumors disappeared in a high percentage of individuals. In humans, Compound I has shown higher activity against ovarian, breast and uterine tumors than known drugs such as tamoxifen. In the tests listed above, isocordoin, cordoin, and 2 ', 4'-dihydroxychalcone showed particularly significant activity. The compounds of the present invention can be conveniently administered orally or by infusion, and are particularly useful for oral administration because natural or synthetic phospholipids form stable, lipophilic complexes with chalcone And medium chain triglycerides and similar additives have also been found to be useful. The dosage of the compounds of the present invention can vary within a wide range, for example from 10 to 300 mg / day, when administered orally. The following examples further illustrate the invention. Example I-Preparation of cordoin xymeninate 7 g of 4-O-prenyl-2-hydroxyacetophenone are reacted with 7 g of benzaldehyde in 10 g of piperidine and 70 ml of ethanol at 60-70 <0> C. After 36 hours, the solvent is removed in vacuo and the residue is dissolved in 100 ml of benzene, which is washed thoroughly with 2N HCl. After removing the benzene, the residue is purified on a silica gel column to give 3.5 g of cordoin. The obtained cordoin is reacted with 3 g of xymenic acid chloride in 20 ml of anhydrous pyridine. The reaction mixture is poured into water and the product is extracted with methylene chloride. After crystallization from methanol, m.p. p. There are obtained 4.2 g of cordoin ximenate having 164-166 ° C. Example II-Preparation of isocordoin palmitate 10 g of 3-C-prenyl res acetophenone and 10 g of p-hydroxybenzaldehyde are dissolved in 15 g of piperidine and 200 ml of absolute ethanol and kept at 60 ° C for 4 hours. . After removal of the solvent, the residue is suspended in 50 ml of 2N HCl and the product is extracted with methylene chloride. After removing the chlorinated solvent, the residue was purified on silica gel, m.p. p. 4.1 g of isocordoin having a temperature of 160-1 ° C. are obtained. The product is reacted with 8 g of palmitoyl chloride in 30 ml of anhydrous pyridine. After diluting the reaction mixture with water and purifying on silica gel, m.p. p. 6.2 g of dipalmitoyl isocordoin having 131-132 ° C. are obtained. Example III-Preparation of acetate and butyrate esters of cordoin and isocordoin These products are prepared according to Examples I and II, using the corresponding acid chloride or anhydride, respectively. (Coldoin acetate mp 131-3 ° C; cordoin butyrate mp 124-6 ° C) Example IV-Preparation of a complex of dipalmitoylphosphatidylcholine and isocordoin 3.08 g of isocordoin was suspended in 30 ml of methylene chloride. 7.9 g of dipalmitoyl phosphatidylcholine are added and left to react for 1 hour with stirring. Upon complete dissolution of these reagents, the reaction mixture is concentrated to a small volume and the concentrate is poured into 50 ml of n-hexane. The precipitated solid material was filtered and dried under vacuum at 40 ° C. overnight, m.p. p. 7.2 g of phospholipid isocordoin complex having a temperature of 70 ° C. are obtained.

[Procedure of Amendment] Article 184-8, Paragraph 1 of the Patent Act [Submission date] November 21, 1996 [Correction contents]                                The scope of the claims 1. Acetic acid, butyric acid, palmitic acid, xymenic acid, isocordoin, cordoy , 4-hydroxyderricin, 2-hydroxyderricin, 3-hydroxyderici 2 ', 4'-dihydroxychalcone, 4,2', 4'-trihydroxychalcone Con ester. 2. An ester of cordoin or isocordoin according to claim 1. 3. Drugs with antagonistic activity against anticancer and type II estrogen receptors Acetic acid, butyric acid, palmitic acid, xymenic acid and isocordoin for preparation Cordoin, 4-hydroxyderricin, 2-hydroxyderricin, 3-hydroxy Sidederin, 2 ', 4'-dihydroxychalcone, 4,2', 4'-trihydro Use of esters of xychalcone. 4. Natural or synthetic phospholipids, isocordoin, cordoin, 4-hydro Xydericin, 2-hydroxyderricin, 3-hydroxyderricin, 2 ', 4' A complex of dihydroxychalcone and 4,2 ', 4'-trihydroxychalcone. 5. In a mixture with a suitable additive, the isocordoin of claim 1-2, Rudoin, 4-hydroxyderricin, 2-hydroxyderricin, 3-hydroxy Delicin, 2 ', 4'-dihydroxychalcone, 4,2', 4'-trihydroxy Contains an ester of sicalcon or the complex of claim 4 as an active ingredient. Formulation composition.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI C07C 69/28 C07C 69/28 // A61K 31/66 A61K 31/66 (81) Designated country EP (AT, BE, CH, DE , DK, ES, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, ML, MR, NE) , SN, TD, TG), AP (KE, LS, MW, SD, SZ, UG), AL, AM, AT, AU, BB, BG, BR, BY, CA, CH, CN, CZ, DE , DK, EE, ES, FI, GB, GE, HU, IS, JP, KG, KP, KR, KZ, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, X, NO, NZ, PL, PT, RO, RU, SE, SG, SI, SK, TJ, TM, TT, UA, US, UZ, VN

Claims (1)

[Claims] 1. Drugs with antagonistic activity against anticancer and type II estrogen receptors Isocordoin, cordoin, 4-hydroxyderricin, 2-h for preparation Droxydelicin, 3-hydroxyderricin, 2 ', 4'-dihydroxychalcone Containing 4,2 ', 4'-trihydroxychalcone and up to 22 carbon atoms. Of saturated or unsaturated linear or branched fatty acids and their esters for. 2. Saturated or unsaturated straight-chain or branched containing up to 22 carbon atoms Fatty acids, isocordoin, cordoin, 4-hydroxyderricin, 2-hydride Roxydelicin, 3-hydroxyderricin, 2 ', 4'-dihydroxychalcone Esters of 4,2 ', 4'-trihydroxychalcone. 3. The claim, wherein the acid is selected from acetic acid, butyric acid, palmitic acid, xymenic acid. The ester according to range 2. 4. The beauty treatment of a cordoin or an isocordoin according to claim 2 or 3. Le. 5. Natural or synthetic phospholipids, isocordoin, cordoin, 4-hydro Xydericin, 2-hydroxyderricin, 3-hydroxyderricin, 2 ', 4' A complex of dihydroxychalcone and 4,2 ', 4'-trihydroxychalcone. 6. In a mixture with appropriate additives, isocordoin, cordoin, 4-hydr Roxydelicin, 2-hydroxyderricin, 3-hydroxyderricin, 2 ', 4 '-Dihydroxychalcone, 4,2', 4'-trihydroxychalcone, or Claims 2-4 esters, or claims A pharmaceutical composition comprising the complex of range 5 as an active ingredient.