JP2006525291A - 1-Aryl-3- (indol-5-yl) prop-2-en-1-one, compositions containing the compounds, and uses thereof - Google Patents

Abstract

The present invention relates to novel compounds, and more particularly to novel 1-aryl-3- (indol-5-yl) prop-2-en-1-ones, compositions containing them, and uses thereof.
The present invention relates to novel substituted 1-aryl-3- (indol-5-yl) prop-2-en-1-ones that are particularly useful for therapeutic activity, especially cancer.
[Chemical 1]

Description

  The present invention relates to novel compounds, and more particularly to novel 1-aryl-3- (indol-5-yl) prop-2-en-1-ones, compositions containing them, and their use as pharmaceuticals. .

  More specifically, the present invention relates to a novel specific 1-aryl-3- (indol-5-yl) prop-2-en-1-one that exhibits anticancer activity, particularly tubulin polymerization inhibitory activity.

  1-Aryl-3- (indol-5-yl) prop-2-en-1-one represents a subclass of chalcones in which the phenyl ring is replaced with indole. In general, chalcones have been extensively described in the literature for over a century. However, although hundreds of publications are aimed at the therapeutic use of chalcones, few mention the use of them in tumors.

  Among the documents describing the use of chalcones for tumors, the following patents and publications can be cited.

  WO 01/72980 discloses substituted chalcones that exhibit anti-cancer and anti-inflammatory activities.

  WO 99/22728 is particularly and generally for the treatment of medical conditions such as alopecia, baldness, obesity, skin disease, prostate cancer and breast cancer in order to suppress the activity of 5α-reductase on steroid hormones. Claims for substituted chalcones.

  WO 99/00114 claims the use of chalcones whose prop-2-en-1-one chain may be saturated or unsaturated.

  WO 98/58913 describes chalcones derived from 1- (2-hydroxyphenyl) -3-aryl-prop-2-en-1-one having growth inhibitory action.

EP 288794-B1 uses 1-aryl-3- (4-X-phenyl) prop-2-en-1-one (where X is a substituent NR ₂ or NHCOR, R = alkyl) for tumors Claims.

  WO 91/17749 claims a cancer treatment using specific chalcones. These chalcones are very generally described and claimed.

  Michael L. Edwards et al. Introduce chalcones that can be used as antimitotic agents in the document "J. Med. Chem. 1990, Vol. 33, pp 1948-1954". Chalcones have been shown and tested in cancer cell lines in vitro.

  Sylvie Ducki et al. Introduces chalcones having anti-mitotic activity in the document “Bioorg. Med. Chem. Letters 1988, vol. 8, pp1051-1056”. Their study was conducted by Michael L., cited above. Based on the work of Edwards et al. The authors observed that replacing 4-N, N-dimethylamino substituents with 4-methoxy and 3-hydroxy substituents significantly improved anti-mitotic activity, especially for K562 cells. Yes.

  However, surprisingly, it has been found that products having 1-aryl-3- (indol-5-yl) prop-2-en-1-one units have high tubulin polymerization inhibitory activity. ing.

  Furthermore, these products induce necrosis very greatly in vivo, which is a highly desirable result for future development of effective drugs for the treatment of cancer.

  Also, with the product of the present invention, tumor necrosis occurs within minutes of test product injection, the tumor center is completely destroyed within a day, and is evident to surrounding normal cells It has been observed that there is no significant effect. Therefore, these products are very risky for surgical resection for inoperable tumor patients, i.e. (i) for the immediate life of the patient, or (ii) for the possible impact on the patient's quality of life. It can be useful in the treatment of patients who give sex (ineffectiveness).

  Furthermore, the products of the present invention are metabolized almost rapidly by the body, limiting long-term effects.

  Another challenge given was to obtain a product having all of the above advantages without suffering the disadvantages often encountered in the study of chalcone products. These disadvantages are (i) the issue of cardiotoxicity, which can be specifically assessed by measuring the inhibition of the Herg receptor, (ii) in the end, the degree is relatively Although it is low, it is a problem of mutagenicity, and this can be specifically evaluated by the Ames test.

  To date, the Applicant has made considerable research efforts in the above fields, but its findings finally identify two very small product classes that satisfy all these criteria simultaneously. I was able to.

ここで、
ａ）Ｙは、ハロゲン、メチルからなる群より選択され、
ｂ）Ａｒは、下記の群から選択され、

ｃ）Ｒは次の通りである：
−ＣＨ₂−ＣＨ₂−ＯＨ。 These products correspond to the following formula (I):

here,
a) Y is selected from the group consisting of halogen, methyl;
b) Ar is selected from the following group:

c) R is as follows:
-CH ₂ -CH ₂ -OH.

  The products of the present invention include isomers. Of the isomers, the Z and E isomers form part of the present invention. The E isomer is preferred.

Preferably Y is CH ₃ .

  Furthermore, in this class, two products can be distinguished due to the additional advantages associated with noticeable properties, in particular solubility problems. This solubility problem is usually only unsatisfactory solved by preparing prodrugs or galenical preparations that can cause serious side effects.

及び
- Ｅ−２−メチル−３−［１−（２−ヒドロキシエチル）−１−Ｈ−インドール−５−イル］−１−（２，５−ジメトキシフェニル）プロペノン：

These two products are:
-E-2-methyl-3- [1- (2-hydroxyethyl) -1-H-indol-5-yl] -1- (3,4,5-trimethoxyphenyl) propenone:

as well as
-E-2-methyl-3- [1- (2-hydroxyethyl) -1-H-indol-5-yl] -1- (2,5-dimethoxyphenyl) propenone:

  The value of the product of the present invention can be advantageously increased by preparing it in the form of a pharmaceutical composition in combination with a pharmaceutically acceptable excipient.

  The products according to the invention can be used for the manufacture of a medicament useful for the treatment of pathological conditions, in particular for the treatment of cancer.

  The invention also relates to a therapeutic composition comprising a compound of the invention in combination with a pharmaceutically acceptable excipient depending on the dosage form selected. The pharmaceutical composition can be provided in solid or liquid form or in the form of liposomes.

  Solid compositions include the described powders, gelatin capsules, and tablets. Oral dosage forms can also include solid forms that are protected from the acidic medium of the stomach. Carriers used in this solid form include inter alia inorganic carriers such as phosphates and carbonates, or organic carriers such as lactose, cellulose, starch or polymers. Liquid forms include solutions, suspensions, or dispersants. They include water or organic solvents (such as ethanol and NMP) or mixtures of surfactants and solvents or mixtures of complexing agents and solvents as dispersible carriers.

  The liquid form is preferably injectable and will therefore have a formulation that is acceptable for such use.

  Acceptable routes of administration by injection include intravenous, intraperitoneal, intramuscular, and subcutaneous routes, with the intravenous route being preferred.

  The dosage of the compound of the invention to be administered is adjusted by the doctor according to the route of administration, the patient, and the patient's condition.

The compounds of the present invention can be administered alone or as a mixture with another anticancer agent. Possible concomitant drugs are shown below.
Alkylating agents, especially cyclophosphamide, melphalan, ifosfamide, chlorambucil, busulfan, thiotepa, predonimustine, carmustine, lomustine, semustine, steptozotocin, decarbazine, temozolomide, procarbazine, and hexamethylmelamine platinum derivatives, especially cisplatin , Carboplatin, oxaliplatin ・ antibiotics, especially bleomycin, mitomycin, dactinomycin ・ antimicrotubules, especially vinblastine, vincristine, vindesine, vinolerubin, taxoid (paclitaxel, docetaxel)
Anthracyclines, especially doxorubicin, daunorubicin, idarubicin, epirubicin, mitoxantrone, rosoxanthroneGroup I and II topoisomerases such as etoposide, teniposide, amsacrine, irinotecan, topotecan, and tomdex fluoropyrimidine, such as 5- Fluorouracil, UFT, Floxuridine-Cytidine analogs such as 5-azacytidine, cytarabine, gemcitabine, 6-mercaptomuline, 6-thioguanine adenosine analogs such as pentostatin, cytarabine, or fludarabine phosphate methotrexate and folinic acid Various enzymes and compounds such as L-asparaginase, hydroxyurea, trans-retinoic acid, suramin, dexrazoki Emissions, amifostine, herceptin, estrogens, androgenic hormones, anti-angiogenesis agents, for example, combretastatin or colchicine derivatives and prodrugs thereof

  The compounds of the present invention can also be combined with radiation therapy. These treatments can be performed simultaneously, separately or sequentially. This treatment will be adapted to the patient being treated by the doctor.

  The product according to the present invention can promote the destruction of cell populations derived from vascular tissue. More particularly, the products of the present invention can be used for initial therapeutic applications that inhibit the growth of cancer cells and simultaneously destroy existing blood vessels. Inhibition of angiogenesis is quantified by the following cell detachment test.

A test that quantifies the detachment of test endothelial cells that allows the quantification of angiogenesis inhibition was developed to select products for "in vitro" activity. This test for quantifying endothelial cell detachment is tested after planting and culturing the endothelial cells in a plate medium, the bottom of which is preferably covered with a binder selected from gelatin, fibronectin or vitronectin The cells are supplemented with a medium containing the compound to be added, the cells are then labeled with a fluorescent substance, the detached cells are washed away, and the fluorescence of the remaining cells is counted with a fluorometer.

  This test preferably consists of measuring the detachment of endothelial cells cultured on a substrate based on a binder selected from fibronectin, vitronectin or gelatin. Preferably, for example, one day after seeding the cells in a plate medium containing 96 wells, the medium is replaced with a medium that contains the compound to be tested and does not contain serum. Similar preparations are made for 6 controls at 3 different concentrations (0.1, 0.3, and 0.6 μM) and 6 controls with no anti-angiogenic product added. Two hours after treatment with the substance to be tested, cells are labeled with calcein-AM (1.6 μg / ml) in medium supplemented with 0.1% BSA. The detached cells are washed away with a medium containing 0.1% bovine serum albumin, and 100 μl of medium is added to each well. Remaining cell fluorescence is counted with a fluorometer. Data obtained are expressed relative to control (untreated cells).

  Evaluation of endothelial cell detachment in vitro is quantified in the following manner. HDECC cells (Human Dermal Microvascular Endothelial Cell (human skin microvascular endothelial cells), Promocell, c-122102) with 5% fetal calf serum, growth factors (EGF 10 ng / ml, hydrocortisone 1 μg / ml, heparinized) Incubate in ECGM-MV medium containing 0.4% growth supplement) and antibiotics (amphotericin 50 ng / ml, gentamicin 50 μg / ml). For detachment testing, 5000 cells HDMEC are planted in clean bottom 96-well plate media (Coster) precoated with fibronectin (10 μg / ml) or vitronectin (1 μg / ml) or gelatin. After 2 hours, the medium is replaced with ECGM-MV 0.1% BSA medium containing the indicated product. Test concentrations are 0.1-0.3 μM and 1 μM for each product. Two hours after treatment, label with calcein (1.6 μg / ml, Molecular Probes) in ECGM-MV 0.1% BSA medium for 1 hour. The detached cells are then washed away with ECGM-MV 0.1% BSA medium and 100 μl of medium is added to each well. The fluorescence of the cells remaining bound to the well substrate is counted using a fluorimeter Spectrafluor Plus (Tecan excitation 485 nm, emission 535 nm). Data is the average of 6 different samples and is expressed as a percentage of the control (untreated cells).

  A cell detachment effect of 15% or more is considered significant.

  The product according to the invention can be useful for inhibiting tubulin polymerization. Inhibition of tubulin can be performed in vitro. Examples of methods for inhibiting tubulin in vitro are described below.

Evaluation of Tubulin Polymerization Inhibition Tubulin was prepared according to the published method “Shelanski et al., Proc. Natl. Acad. Sci. USA, 70, 765-768. Weingarten et al., 1975, Proc. Natl. Acad. Sci. USA, 72, 1858-1862 "purified from porcine brain. Briefly, the brain is ground and centrifuged in extraction buffer. Tubulin contained in the supernatant of the extract was polymerized at 37 ° C. and separated from MAPs (Microtubule Associated Protein) by chromatography using a phosphocellulose P11 column (Whatman). It is subjected to two successive cycles of depolymerization at ° C. The tubulin thus separated has a purity of more than 95%. This was stored in a buffer of 30% glycerol with the designation RB / 2, the composition of which is as follows: 50 mM MES-NaOH [2- (N-morpholino) ethanesulfonic acid], pH 6.8; 0.25 mM MgCl ₂ ; 0.5 mM EGTA; 30% (v / v) glycerol, 0.2 mM GTP (guanosine-5′-triphosphate).

Tubulin polymerization into microtubules is monitored by nephelometry as follows: 10 μM of tubulin in 30% glycerol buffer in RB / 2 supplemented with 1 mM GTP and 6 mM MgCl _2. 1 mg / ml). Polymerization is induced by raising the temperature from 6 ° C. to 37 ° C. in a 1 cm path length cuvette placed in a UVIKON 931 spectrophotometer (Kontron) equipped with a temperature controlled cuvette holder. The increase in turbidity of the solution is monitored at 350 nm.

The product is dissolved 10 mM in DMSO and added to the tubulin solution at various concentrations (0.5-10 μM) before polymerization. IC ₅₀ is defined as the concentration of product that inhibits polymerization at a rate of 50%. Products with an IC ₅₀ of 3 μM or less are considered very active.

Evaluation of growth inhibition of Hela tumor cells or endothelial cells HDMEC Proliferation of Hela or HDMEC cells is assessed by measuring ¹⁴ C thymidine incorporation as follows. Hela cells (human-derived epithelial tumor cells) are cultured in DMEM medium (Gibco) containing 10% fetal bovine serum and antibiotics (1% penicillin, 1% streptomycin). To perform a proliferation test, cells are seeded in 96-well Cytostar microplates (Amersham) at a rate of 5000 cells per well. ¹⁴ C thymidine (0.1 μCi / well) and the product to be evaluated are then added. The product is used at various concentrations below 10 μM and DMSO (the solvent used to solubilize the product) should not exceed 0.5% in the medium. After 48 hours in culture at 37 ° C., the radioactivity incorporated into the cells is measured by counting the plate medium in a TRI-LUX counter (Wallac). IC ₅₀ is defined as the concentration of product at which the radioactivity is reduced by 50% compared to the untreated control. Products with an IC ₅₀ of less than 1 μM are considered cytotoxic.

Evaluation of tumor necrosis in vivo Mice were obtained from species obtained from Jackson Laboratories (Bar Harbor, ME, USA) by IFFA-CREDO (Domaines des Incins, 69210 L'Arbresle, France) or from Charles River (USA). The seeds are raised by Charles River France (76410 St Aubinles Elbeuf, France). The mice initially weigh more than 18 g at the start of the test. Mice have free access to food (UAR reference 113, Villemoisson, 91160 Epinay sur Org, France) and water.

  The tumor used is currently transplanted into our laboratory. All of these tumors are from the National Cancer Institute (NCI) Frederick Cancer Research Facility (Fredrick, MD, USA), or the American Type Culture Collection (ATCC, Rockville, MD), US.

  Tumor transplantation, chemotherapy, and data analysis techniques have been published in detail (Corbett et al., 1982a; Corbett et al., 1982b).

  In summary, the animals required for one experiment are collected and transplanted bilaterally at 0 (zero) days. Solid tumor growth proceeds freely to the desired size. The mice are then treated with an intravenous injection of the test compound in solution.

  Tumor sample collection is usually (but not essential) performed 24 hours after treatment.

  Displace the brain to kill the mouse. The transplanted tumor, the overlying skin, and surrounding tissue are collected and stored in 10% (v / v) formaldehyde (Carlo Erba, Val de Reuil, France).

Samples are then treated, cut into sections, stained with hematoxylin, eosin, and saffron yellow, and then examined visually. Tumor necrosis (necrosis ± degeneration) is assessed by microscopic examination using grades ranging from 0 to 5.
0 = no necrosis 1 = very little <5%
2 = low 5-25%
3 = Moderate 25-50%
4 = remarkable 50-75%
5 = Large amount> 75%

  The values given for tumor necrosis 24 hours after administration of the test compound correspond exclusively to necrosis depending on the product, which should be clearly distinguished from all existing necrosis resulting from the experiment. Can do.

  Experimental necrosis was assessed based on untreated controls.

  The tumor model is C51 murine adenocarcinoma. The colon tumor is a grade III mucosal colon adenoma. This is maintained in female BALB / c mice by successive subcutaneous passages every 18 days. Experiments were performed on female BALB / c mice.

The following results were obtained for the following examples under the conditions described in the results.
Example 2:
Selected dosing: 35 mg / kg-Grade 5 necrosis

References CORBETT, T.W. H. , LEOPOLD, W.M. R. , DYKES, D.M. J. et al. , ROBERTS, B.R. J. et al. , GRISWOLD, D.M. P. , Jr. and SCABEL, F.A. M.M. , Jr. , Toxicity and anticancer activity of a new triazine antiflate (NSC 127755). Cancer Res. 1982a, 42, 1707-1715.
CORBETT, T.W. H. , ROBERTS, B.R. J. et al. , TRADER, M .; W. , LASTER, W.M. R. , Jr. , GRISWOLD, D.M. P. , Jr. and SCABEL F. M.M. , Jr. , Response of trans- plantable tumors of mice to anthracene derivatives alone and in combination with clinically used agents. Cancer Treat. Rep. 1982b, 66, 1187-11200.

The definition “halogen” is an element selected from F, Cl, Br, and I.

（ここに、Ａｒは先に定義されたものであり、Ｙはハロゲン原子と異なるものであり、ＲはインドールのＮＨ官能基を保護する基である）は、一般式（II）の芳香族ケトン（Ｙはハロゲン原子と異なるものである）と一般式（III）の芳香族アルデヒドを、スキーム（Ｉ）にしたがい「Ｊ．Ｍｅｄ．Ｃｈｅｍ．，１９９０，３３，１９４８」に記載の条件下で結合させることによって製造することができる。

Chalcones of general formula (I):

(Wherein Ar is as defined above, Y is different from a halogen atom, and R is a group protecting the NH functional group of indole) is an aromatic ketone of the general formula (II) (Y is different from a halogen atom) and an aromatic aldehyde of the general formula (III) bonded under the conditions described in “J. Med. Chem., 1990, 33, 1948” according to scheme (I). Can be manufactured.

  This operation is generally performed in a Soxhlet-type apparatus at the reflux temperature of an alcohol such as ethanol in the presence of piperidine, acetic acid, and molecular sieves.

  It is understood that the coupling between the ketone of general formula (II) and the aldehyde of general formula (III) can be carried out using protecting groups such as trimethylsilyl, triethylsilyl, or tert-butyldimethylsilyl. The attachment and cleavage of the protecting group for the NH function of the indole can be performed under the conditions described in “Protective Groups in Organic Chemistry (Wiley publisher)”.

  Aromatic aldehydes of general formula (III) are commercially available or have already been described in the literature.

  Aromatic ketones of general formula (II) are described in the literature and can generally be prepared from commercially available corresponding aromatic aldehydes. When Y is a methyl group, the procedure is to react the aldehyde with an appropriately selected organometallic reagent and then obtain the benzyl obtained under the conditions described in “J. Med. Chem., 1990, 33, 1948”. This can be done successfully by oxidizing the alcohol.

（ここに、Ａｒは上記の通りであり、Ｒはハロゲン交換の条件下で置換されない置換基、Ｙはハロゲン原子、好ましくは臭素又はヨウ素原子である）は、ハロゲンの付加とその後のカルコンの脱ハロゲン化水素反応によって製造することができ、ここで、Ｙはスキーム（II）においてハロゲン原子である。インドールの窒素を例えばトリメチルシリル、トリエチルシリル、ｔｅｒｔ−ブチルジメチルシリル、又はｔｅｒｔ−ブチルオキシカルボニルによって保護する形態でこの一連の付加・脱離を行い、次いで脱保護することは、とりわけ有益である。Ｙがフッ素の場合、当業者には公知のように、シリル化された保護基の使用は推奨されない。 Chalcones of general formula (I):

(Where Ar is as described above, R is a substituent that is not substituted under halogen exchange conditions, and Y is a halogen atom, preferably a bromine or iodine atom) is the addition of halogen followed by the removal of chalcone. It can be produced by a hydrogen halide reaction, where Y is a halogen atom in scheme (II). It is particularly beneficial to perform this series of additions and removals followed by deprotection in the form of protecting the indole nitrogen with, for example, trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, or tert-butyloxycarbonyl. When Y is fluorine, the use of silylated protecting groups is not recommended as is known to those skilled in the art.

  The addition of halogen, preferably bromine or chlorine, is generally carried out in a solvent such as chloroform or carbon tetrachloride at a temperature of 0-50 ° C. Dehydrohalogenation is generally carried out in a solvent such as dichloromethane in the presence of an organic or inorganic base such as triethylamine or sodium hydroxide or potassium carbonate at a temperature from 0 ° C. to the reflux temperature of the reaction medium. This dehydrohalogenation can be accompanied by at least partial deprotection of the indole nitrogen, in particular in the case of the tert-butyloxycarbonyl group, depending on the protecting group used.

（ここに、Ａｒ、Ｒ、及びＹは本発明に一致する）は、置換基Ｒが水素原子である一般式（Ｉ）のカルコンのアルキル化によって製造することができる。この反応は、２つの段階で行われる。第１の段階において、例えば、ＮａＨ、ＬｉＨ、又はＫＨのようなアルカリ金属水素化物を利用してアルカリ金属塩の形態でインドールの窒素を活性化した後、２−ブロモエトキシ−ｔｅｒｔ−ブチルジメチルシランのようなハロゲン化アルキルを、Ｒが２−ｔｅｒｔ−ブチルジメチルシリルオキシエチルである一般式（Ｉ）の生成物を生成させるために反応させる。第２の段階は、テトラ−Ｎ−ブチルアンモニウムフルオリドのようなアルカリ金属フッ化物を利用して、トリメチルシリル保護基を開裂させ、Ｒが−ＣＨ₂ＣＨ₂ＯＨである本発明の生成物を生成させることからなる。 Chalcones of general formula (I):

(Wherein Ar, R and Y are consistent with the present invention) can be prepared by alkylation of chalcones of general formula (I) wherein the substituent R is a hydrogen atom. This reaction takes place in two stages. In the first step, after activation of the indole nitrogen in the form of an alkali metal salt using, for example, an alkali metal hydride such as NaH, LiH or KH, 2-bromoethoxy-tert-butyldimethylsilane Are reacted to produce a product of general formula (I) wherein R is 2-tert-butyldimethylsilyloxyethyl. The second step utilizes an alkali metal fluoride such as tetra-N-butylammonium fluoride to cleave the trimethylsilyl protecting group to produce the product of the invention where R is —CH ₂ CH ₂ OH. Made up of.

〔Example〕
The following examples are presented to illustrate the present invention.

Example 1 : E-2-methyl-3- [1- (2-hydroxyethyl) indol-5-yl] -1- (3,4,5-trimethoxyphenyl) propenone

Step 1 : 2.24 g of 1- (3,4,5-trimethoxyphenyl) propanone ("Biorg. Med. Chem. 1998, 8 (9), 1051") in 100 ml of ethanol containing 2 ml of piperidine and 1 ml of acetic acid. Thus, 2.76 g of 1-tert-butyloxycarbonylindole-5-carboxaldehyde (which can be prepared according to “J. Org. Chem. 2002, 67 (17), 6256-59”) The flask was sequentially placed in a 25 ml three-necked flask equipped with a sieve filled soxhlet. The reaction medium was heated under reflux for 48 hours. After cooling, the reaction medium is concentrated under reduced pressure and then taken up in 100 ml of ethyl acetate, the organic phase is washed with water, dried over magnesium sulphate and concentrated under reduced pressure. The crude product is purified by flash chromatography on silica gel (70-230 mesh) and elution with a mixture of cyclohexane and ethyl acetate (volume ratio 70/30), in the form of a pale yellow oil. 2 g of pure E-2-methyl-3- [1- (1-tert-butyloxycarbonyl) -1-H-indol-5-yl] -1- (3,4,5-trimethoxyphenyl) propenone Got. This was used as such for the next step.

Step 2 : 0.7 g E-2-methyl-3- [1- (1-tert-butyloxycarbonyl) -1-H-indol-5-yl] -1- (3,4,5-trimethoxy Phenyl) propenone was dissolved in 15 ml of THF. Subsequently, 1.5 ml of methanol and 0.25 g of sodium methoxide were added successively, and then the reaction medium was stirred at room temperature for 18 hours. After concentration under reduced pressure, the reaction medium is taken up in 75 ml of ethyl acetate and 35 ml of water. The organic phase was separated by decantation, washed with water, dried over magnesium sulfate and concentrated under reduced pressure. After purification by flash chromatography on silica gel (70-230 mesh) and elution with a mixture of dichloromethane and diisopropyl ether (volume ratio 50/50), 505 mg of pure, in the form of an orange oil. E-2-methyl-3- (1-H-indol-5-yl) -1- (3,4,5-trimethoxyphenyl) propenone was obtained. The characteristics were as follows:
Mass spectrum (EI): m / z = 351
Elemental analysis: C = 71.26%, H = 6.54%, N = 3.72%

Step 3 : 1.4 g of E-2-methyl-3- (1-H-indole-) obtained in Step 2 in 40 ml of pyridine cooled to 0 ° C. with 360 mg of sodium hydride (60% in oil) Slowly added to a solution of 5-yl) -1- (3,4,5-trimethoxyphenyl) propenone. After stirring for 1 hour at 0 ° C. until gas evolution ceased, 960 mg of 2-bromoethoxy-tert-butyldimethylsilane was added and the reaction medium was heated at 60 ° C. for 2 hours. After adding 80 ml of water, the reaction medium was extracted three times with 50 ml of ethyl acetate. The combined organic phases were washed with water, dried over magnesium sulfate and concentrated to dryness under reduced pressure. The resulting brown oily residue was purified by flash chromatography on silica gel (70-230 mesh) and elution with a mixture of dichloromethane and diisopropyl ether (volume ratio 50/50). In this way 1.5 g of E-2-methyl-3- [1- (2-tert-butyldimethylsilyloxyethyl) indol-5-yl] -1- (3,4,5-trimethoxyphenyl) Propenone was obtained in the form of a yellow oil, which was used as such in the next step.

Step 4 : 1.5 g E-2-methyl-3- [1- (2-tert-butyldimethylsilyloxyethyl) indol-5-yl] -1- (3,4,5- A solution of trimethoxyphenyl) propenone was stirred for 48 hours at room temperature with 23.5 ml of a 1M solution of tetra-N-butylammonium fluoride in tetrahydrofuran. After adding 25 ml of water, the reaction medium was extracted 3 times with 5 ml of ethyl acetate. The combined organic phases were washed with water, dried over magnesium sulfate and concentrated to dryness under reduced pressure. The orange residue was purified by flash chromatography on silica gel (70-230 mesh), elution with a mixture of dichloromethane and ethanol (volume ratio 95/5) and further crystallization from isopropanol. There were thus obtained 750 mg of E-2-methyl-3- [1- (2-hydroxyethyl) indol-5-yl] -1- (3,4,5-trimethoxyphenyl) propenone, which has the following characteristics: It was as follows.
Mass spectrum (EI): m / z = 395
Elemental analysis: C = 69.72%, H = 6.25%, N = 3.53%
Melting point (Kofler) = 119 ° C.

Example 2 : E-2-methyl-3- [1- (2-hydroxyethyl) indol-5-yl] -1- (3-methoxy-4,5-methylenedioxyphenyl) propenone

Step 1 : In a 250 ml three-necked flask under an argon atmosphere, 5 g of indole-5-carboxaldehyde was dissolved in 90 ml of DMF and 18 ml of DMSO, and then the reaction medium was cooled to 0 ° C. Then, 2.06 g of 60% sodium hydride in oil was added in portions, and then the reaction medium was stirred until gas evolution ceased while returning to room temperature. Then 8.6 g of (2-trimethylsilylethyl) oxymethyl chloride was poured dropwise and the reaction medium was stirred at room temperature for 20 hours. The reaction medium was then poured onto a mixture of 300 ml of water and 100 g of crushed ice and extracted three times with 150 ml of ethyl acetate. The combined organic phases were washed with a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and concentrated to dryness under reduced pressure. The resulting brown oil was purified by flash chromatography on silica gel (70-230 mesh) and elution with a mixture of cyclohexane and ethyl acetate (volume ratio 70/30). In this way 9 g of 1- (2-trimethylsilylethyl) oxymethylindole-5-carboxaldehyde was obtained in the form of an orange oil, which was used as such in the next step.

Stage 2 : 2.1 g of 1- (3-methoxy-4,5-methylenedioxyphenyl) propanone (which can be prepared according to “J. Org. Chem. 1981, 46 (14), 2969-71”); Soxhlet filled with 3Å molecular sieves over 96 hours with 2.76 g 1- (2-trimethylsilylethyl) oxymethylindole-5-carboxaldehyde in 100 ml ethanol containing 2 ml piperidine and 1 ml acetic acid. It put into the placed 250 ml three neck flask sequentially. After cooling, the reaction medium is concentrated under reduced pressure, taken up in 100 ml of ethyl acetate, the organic phase is washed with water, dried over magnesium sulphate and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel (70-230 mesh), elution with a mixture of cyclohexane and ethyl acetate (volume ratio 70/30) and crystallization from diisopropyl ether. 2 g of pure E-2-methyl-3- [1- (2-trimethylsilylethyl) oxymethyl-1-H-indol-5-yl] -1- (3-methoxy-4,5 in the form of white crystals -Methylenedioxyphenyl) propenone was obtained. This characteristic was as follows.
Melting point (Kofler) = 90 ° C.

Step 3 : 2 g E-2-methyl-3- [1- (2-trimethylsilylethyl) oxymethyl-1-H-indole-5 in 42 ml THF in a 250 ml 3-neck flask under argon atmosphere Yl] -1- (3-methoxy-4,5-methylenedioxyphenyl) propenone, then 4.3 ml of a 1M solution of tetra-N-butylammonium fluoride in THF is added and the reaction medium is refluxed Heated under 20 hours. After concentration under reduced pressure, the reaction medium is taken up in 75 ml of ethyl acetate and 75 ml of water. The organic phase was separated by decantation, washed with water, dried over magnesium sulfate and concentrated to dryness under reduced pressure. The resulting red oil was purified by flash chromatography on silica gel (70-230 mesh), elution with a mixture of cyclohexane and ethyl acetate (volume ratio 70/30) and further crystallization from isopropanol. In this way in the form of a beige solid, 420 mg of pure E-2-methyl-3- (1-H-indol-5-yl) -1- (3-methoxy-4,5-methylenedioxy Phenyl) propenone was obtained. This characteristic was as follows.
Melting point (Kofler) = 140 ° C.

Step 4 : Following the procedure of Step 3 of Example 1, except that 671 mg of E-2-methyl-3- (1-H-indol-5-yl) -1- (3 Starting from -methoxy-4,5-methylenedioxyphenyl) propenone, 180 mg 60% sodium hydride in oil, 479 mg 2-bromoethoxy-tert-butyldimethylsilane in 20 ml pyridine, silica gel (70-230 mesh) After purification by flash chromatography above and elution with a mixture of ethyl acetate and cyclohexane (volume ratio 30/70), 830 mg of E-2-methyl-3- [1- (2-tert-Butyldimethylsilyloxyethyl) indol-5-yl] -1- (3-methoxy-4,5-methylenedioxy Phenyl) propenone was obtained. This was used as such for the next step.

Step 5 : The procedure of Step 4 of Example 1 was followed for 20 hours, except that 830 mg of E-2-methyl-3- [1- (2-tert-butyldimethylsilyloxyethyl) indole-in 25 ml of tetrahydrofuran Starting with 13.5 ml of a 1M solution of 5-yl] -1- (3-methoxy-4,5-methylenedioxyphenyl) propenone and tetrabutylammonium fluoride in tetrahydrofuran on silica gel (70-230 mesh) After purification by flash chromatography on silica gel, elution with a mixture of ethyl acetate and cyclohexane (volume ratio 50/50) and crystallization from isopropyl ether, 385 mg of E-2 in the form of a beige powder -Methyl-3- [1- (2-hydroxyethyl) indole-5 Yl] -1- (3-methoxy-4,5-methylenedioxyphenyl) propenone was obtained. This characteristic was as follows.
Mass spectrum (EI): m / z = 379
Elemental analysis: C = 69.84%, H = 6.50%, N = 3.62%
Melting point (Kofler) = 83 ° C.

Example 3 : E-2-methyl-3- [1- (2-hydroxyethyl) -1-H-indol-5-yl] -1- (2,5-dimethoxyphenyl) propenone

Step 1 : 6.13 g of 1-tert-butyloxycarbonylindole-5-carboxaldehyde in 280 ml of ethanol containing 4.86 g of 1- (2,5-dimethoxyphenyl) propanone and piperidine 5 ml and acetic acid 2.53 ml (Manufactured according to “J. Org. Chem. 2002, 67 (17), 6256-59”) was heated for 3 hours with heating under reflux for 48 hours. Sequentially added to the neck flask. After cooling, the reaction medium is concentrated under reduced pressure and then taken up in 100 ml of ethyl acetate, the organic phase is washed with water, dried over magnesium sulphate and concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel (70-230 mesh) and elution with a mixture of cyclohexane and ethyl acetate (volume ratio 90/10). 2.3 g of pure E-2-methyl-3- [1- (1-tert-butyloxycarbonyl) -1-H-indol-5-yl] -1- (2,5- in the form of a yellow oil Dimethoxyphenyl) propenone was obtained and used as such in the next step.

Step 2 : 2.3 g of E-2-methyl-3- [1- (1-tert-butyloxycarbonyl) -1-H-indol-5-yl] -1- (2,5-dimethoxyphenyl) propenone Was dissolved in 57 ml of tetrahydrofuran. Then 5.7 ml of methanol and 0.73 g of sodium methoxide were added sequentially and the reaction mixture was stirred at room temperature for 20 hours. After concentration under reduced pressure, the reaction medium was taken up in 75 ml of ethyl acetate and 35 ml of water. The organic phase was separated by decantation, washed with water, dried over magnesium sulfate and concentrated under reduced pressure. After purification by flash chromatography on silica gel (70-230 mesh) and elution with a mixture of ethyl acetate and cyclohexane (volume ratio 30/70) 1.5 g of pure in the form of a yellow paste E-2-methyl-3- (1-H-indol-5-yl) -1- (2,5-dimethoxyphenyl) propenone was obtained. This characteristic was as follows.
Mass spectrum (EI): m / z = 321
Melting point (Kofler) = 52-55 ° C.

Step 3 : Followed the procedure of Step 3 of Example 1, except that 1 g of E-2-methyl-3- (1-H-indol-5-yl) -1- (2,5-dimethoxyphenyl) propenone, Starting with 280 mg of 60% sodium hydride in oil and 750 mg of 2-bromoethoxy-tert-butyldimethylsilane in 30 ml of pyridine, flash chromatography on silica gel (70-230 mesh), ethyl acetate and cyclohexane After purification by elution with a mixture of (volume ratio 30/70), 1.45 g of E-2-methyl-3- [1- (2-tert-butyldimethylsilyloxyethyl) in the form of a brown oil ) Indol-5-yl] -1- (2,5-dimethoxyphenyl) propenone was obtained. This was used as such for the next step.

Step 4 : The procedure of Step 4 of Example 1 was followed for 20 hours, except that 1.45 g E-2-methyl-3- [1- (2-tert-butyldimethylsilyloxyethyl) indole in 45 ml tetrahydrofuran. Flash chromatography on silica gel (70-230 mesh) starting with 25 ml of a 1M solution of -5-yl] -1- (2,5-dimethoxyphenyl) propenone and tetra-N-butylammonium fluoride in tetrahydrofuran. After purification by chromatography, elution with a mixture of ethyl acetate and cyclohexane (volume ratio 30/70) and further crystallization from isopropanol, 610 mg of E-2-methyl-3- [1- (2-Hydroxyethyl) indol-5-yl] -1- (2,5-dimethoxy Eniru) propenone was obtained. This characteristic was as follows.
Mass spectrum (EI): m / z = 365
Melting point (Kofler) = 112 ° C.

Claims (11)

Formula (I):

(Where
a) Y is selected from the group consisting of halogen and methyl;
b) Ar is the following group:

Selected from
c) R is —CH ₂ —CH ₂ —OH)
And the isomers thereof. The product of claim 1 wherein Y is CH ₃ . Following formula:

E-2-methyl-3- [1- (2-hydroxyethyl) -1-H-indol-5-yl] -1- (3,4,5-trimethoxyphenyl) propenone represented by: Following formula:

E-2-methyl-3- [1- (2-hydroxyethyl) -1-H-indol-5-yl] -1- (2,5-dimethoxyphenyl) propenone represented by: Following formula:

E-2-methyl-3- [1- (2-hydroxyethyl) -1-H-indol-5-yl] -1- (3,4,5-trimethoxyphenyl) propenone represented by:   A pharmaceutical composition comprising the product of any one of claims 1-5 in combination with a pharmaceutically acceptable excipient.   Use of the product according to any one of claims 1 to 6 as a tubulin polymerization inhibitor.   Use of a product according to any one of claims 1 to 6 for promoting the detachment of endothelial cells forming the blood vessel wall supplying the tumor.   Use of the product according to any one of claims 1 to 6 for promoting tumor necrosis.   Use of a product according to any one of claims 1 to 6 for the manufacture of a medicament useful for the treatment of a pathological condition.   Use according to claim 10, wherein the pathological condition is cancer.