WO2009031708A1

WO2009031708A1 - Coumarin compound and use thereof

Info

Publication number: WO2009031708A1
Application number: PCT/JP2008/066486
Authority: WO
Inventors: Hideki Kubo; Hirohisa Nagahori; Yoshitaka Tomigahara; Junya Takahashi
Original assignee: Sumitomo Chemical Company, Limited
Priority date: 2007-09-05
Filing date: 2008-09-05
Publication date: 2009-03-12
Also published as: JP2009062300A

Abstract

Disclosed are: a type-3 17β-hydroxysteroid dehydrogenase-inhibiting composition which is characterized by comprising a coumarin compound represented by the formula (I) or the like and an inert carrier; and others.

Description

Specification

Coumarin compounds and their uses

Technical field

The present invention relates to a coumarin compound and its use. Background art

Prostate cancer, benign prostatic hypertrophy, acne, seborrhea, hirsutism, androgenetic alopecia, premature sexual maturity, adrenal hypertrophy and polycystic ovary syndrome, such as onset and progression Promoted by androgenic activity.

Type 3 1 7 / 3—Hydroxysteroid dehydrogenase is a low activity male hormone

It is an enzyme that catalyzes the conversion to testosterone, a highly active male hormone. For example, in cancerous prostate tissue, it has been observed that the expression level of type 3 1 7] 3-hydroxycysteide dehydrogenase gene is higher than that in normal tissues (eg, The Pros tate, 53, 154-159, (2002)). Disclosure of the invention

An object of the present invention is to provide a compound that can inhibit type 3 17 i3-hydroxysteride dehydrogenase, and thus is useful in the treatment or prevention of male hormone-dependent diseases.

Specifically, the present invention relates to compounds represented by the following formulas (I) and (Π) having the ability to inhibit type 3 17; 3-hydroxysteroid dehydrogenase (hereinafter collectively referred to as the present compound). ).

More specifically, the present invention provides:

1. Formula (I)

[In the formula, I, is a hydrogen atom, a methoxymethyl group, a phenyl group, a benzyl group or fluorine. Represents an optionally substituted CI-C4 alkyl group, represents a hydrogen atom, a halogen atom, a CI-C4 alkyl group, a methoxymethyl group or a phenyl group, and represents a hydrogen atom or a halogen atom. And represents a hydrogen atom, a C1-C4 alkylcarbonyl group or a C1-C4 alkylaminocarbonyl group. However, I, is a hydrogen atom or a methyl group, and Y! When is a hydrogen atom, R! Does not become a hydrogen atom. ]

A type 3 17β-hydroxysteroid dehydrogenase inhibitor composition comprising a coumarin compound represented by formula (II) and an inert carrier;

2. a coumarin compound represented by formula (II);

3. A type 3 1 7 3-hydroxysteroid dehydrogenase inhibitor composition comprising the compound according to item 2 and an inert carrier;

4.3 use of a compound contained as an active ingredient in the composition according to item 1 as an active ingredient for inhibiting type 3 1 7] 3-hydroxysteroid dehydrogenase;

5. Type 3 1 7) Use of the compound according to item 2 as an effective component for inhibiting 3-hydroxysteroid dehydrogenase;

6. A method of inhibiting type 3 1 7 / 3-hydroxysteroid dehydrogenase, wherein the method inhibits a therapeutically effective amount of type 3 1 7 / 3-hydroxystero dedehydrogenase according to item 1 above Administering a coumarin compound contained in the composition as an active ingredient to a patient in need of such inhibition;

7. A method for treating or preventing an androgen-dependent disease, said method comprising a therapeutically effective amount of type 3 17 β according to the preceding paragraph 1 for a patient in need of said treatment or prevention. A method comprising the step of administering a coumarin compound contained as an active ingredient in a hydroxysterol dehydrogenase inhibitor composition; and

8. Male hormone-dependent diseases include prostate cancer, benign prostatic hyperplasia, prostatic intraepithelial neoplasia, hirsutism, acne, seborrhea, androgenetic alopecia, sexual prematurity, adrenal hypertrophy or polycyst The method according to item 7 above, which is ovarian ovary syndrome;

Etc. are provided. According to the present invention, a composition for treating or preventing a male hormone-dependent disease, such as by inhibiting the activity of type 3 17 β-hydroxysteride dehydrogenase and suppressing the enhancement of male hormone activity in tissues, etc. Development and provision are possible. BEST MODE FOR CARRYING OUT THE INVENTION

In the present invention, examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

In the present invention, examples of the CI-C4 alkyl group include a methyl group, an ethyl group, an isopropyl group, a t-butyl group, and the like. Examples of the C 卜 C4 alkylcarbonyl group include a acetyl group and a propionyl group. Examples of the C 卜 C4 alkylaminocarbonyl group include a methylaminocarbonyl group. The compounds also represent their pharmacologically acceptable salts. The pharmacologically acceptable salt represents a salt of the present compound with an inorganic acid, a salt with an organic acid, a salt with an inorganic base or a salt with an organic base. Examples of salts with inorganic acids include hydrochlorides and hydrobromides, and examples of salts with organic acids include acetates and benzoates, and salts with inorganic bases. Examples thereof include potassium salts and sodium salts. Examples of salts with organic bases include pyridine salts and morpholine salts. Among these compounds, compounds (A) represented by compound numbers (1) to (2 4) are exemplified in Tables 1 to 2.

This compound has the ability to inhibit the activity of type 3 17 / 3-hydroxysteride dehydrogenase. This ability is important to prevent the onset and progression of male hormone-dependent diseases by suppressing the production of testosterone, a male active male hormone, leading to decreased male hormone activity in tissues. Thus, the present compound treats or prevents male hormone-dependent diseases by inhibiting type 3 17 / 3-hydroxysteroide dehydrogenase activity and suppressing the enhancement of male hormone activity in tissues. It can be used as an active ingredient in other compositions (pharmaceuticals, cosmetics, food additives, etc.).

For example, the compound is a method of inhibiting type 3 17] 3-hydroxysteroid dehydrogenase, wherein a therapeutically effective amount of at least one compound is administered to a patient in need of such inhibition. It can utilize for the method including the process of administering to. Also for example, the compound is a method of treating or preventing a male hormone dependent disease, comprising the step of administering a therapeutically effective amount of at least one compound to a patient in need of the treatment or prevention. It can be used for the method of inclusion. This compound is useful for the prevention or treatment of male hormone-dependent diseases whose onset or progression is promoted by the activity of male hormones. Male hormone-dependent diseases to which this compound can be applied include, for example, prostate fistula and other androgen-dependent neoplasms, benign prostatic hypertrophy, prostatic intraepithelial neoplasia, androgenetic alopecia (ie male patients) Pattern baldness in both female and female patients), hirsutism, polycystic ovary syndrome, premature sexual maturity, adrenal hypertrophy, seborrhea and acne. For example, treat or prevent prostate cancer, benign prostatic hypertrophy, prostatic intraepithelial neoplasia, hirsutism, acne, seborrhea, androgenetic alopecia, sexual prematurity, adrenal hypertrophy or polycystic ovary syndrome It can be used in a method comprising administering to a patient in need of such treatment or prevention a therapeutically effective amount of at least one compound. A preferred dosage (effective amount) is about 0.001 to 50 O mg of the compound per kg body weight per day. A more preferred dosage is about 0.01 to 25 mg of the present compound or a pharmaceutically acceptable salt of the present compound or a solvate of the present compound per day per kilogram of body weight. The compound is usually administered in the form of a composition comprising at least one compound, or a pharmaceutically acceptable salt of the compound or a solvate of the compound, and at least one inert carrier. The present compounds contained in these compositions are usually from 0.01% by weight to 99.99% by weight, and the inert carrier is usually from 99.99% by weight to 0.9% by weight. 0 1% by weight. The inert carrier is a pharmaceutically acceptable carrier or excipient. The composition of the present invention may further contain a pharmaceutical additive, a cosmetic additive, a food additive and the like. Pharmaceutically acceptable carriers, excipients, pharmaceutical additives, food additives, cosmetic additives, etc. used to prepare the above composition containing the present compound depend on the specific use of the composition. Can be selected as appropriate. Also, the form of the composition can be used for specific purposes. Depending on the way, it can be in the form of various solids, liquids, etc. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. The powder and disintegrant can be comprised of from about 5 to about 95 percent active ingredient. Examples of the solid carrier include magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and methods of manufacture for various compositions are described in A. Gennaro (ed., Remington's Pharmaceutical Sciences, 18th edition, (1990), Mack Publishing Co., Easton, Penn sylv an ia Liquid form preparations include solutions, suspensions, emulsions, etc. For example, for parenteral injection Water or water-propylene glycol solution of the following: Sweeteners and opacifiers may be added to oral solutions, suspensions, and emulsions Examples of liquid form preparations are intranasal. It may also be a liquid formulation for administration, or an aerosol formulation suitable for inhalation, in which case it may include a solution and a solid in powder form. Is a drug such as inert compressed gas (eg nitrogen) Solid form preparations intended to be converted to liquid form preparations, either prior to use, for either oral or parenteral administration Such liquid forms include solutions, suspensions, and emulsions The compounds may be administered transdermally This transdermal composition comprises creams, mouths For this purpose it can be in the form of a cisyon, aerosol and Z or emulsion In addition, it may be contained in a matrix-type transdermal patch or a reservoir-type transdermal patch as usual in the art.

The compound may also be delivered subcutaneously. More preferably, the compound may be administered orally. The composition can also be in unit dosage form. In such form, the composition is subdivided into suitably sized unit doses containing appropriate quantities of the active component, eg, an effective amount to achieve the desired purpose. The amount of active compound in a unit dose of the composition is adjusted to be from about 1 mg to about 100 mg, preferably from about 1 mg to about 50 mg, more preferably from about 1 mg to about 25 mg, according to the particular application. May be. The actual dosage used may vary depending on the weight of the patient and the severity of the disease being treated. If necessary for convenience, the total daily dosage may be divided into several doses. The dosage and frequency of administration of the compound and / or pharmaceutically acceptable salt of this compound will depend on factors such as the patient's age, condition and physique, and the severity of the condition being treated. Consider and adjust according to the judgment of the attending physician. A typical recommended daily dosage regimen for oral administration can range from about lmg to about 500 mg per day, preferably from about lmg to about 20 Omg per day. These can be administered in 2 to 4 divided doses. The present invention includes a therapeutically effective amount of at least one compound, or a pharmaceutically acceptable salt of the compound or a solvate of the compound, and a pharmaceutically acceptable carrier, excipient or A kit comprising a diluent is also provided. Such a kit is one of the kits. When the present compound that can contain the above components in the above container is used by adding it to cosmetics, specific examples of cosmetics to which the compound is added include, for example, liquid, milky, cream, lotion, ointment, Gels, aerosols, mousses and the like can be mentioned. Lotions can be manufactured according to conventional methods using cosmetic additives such as suspending agents, emulsifying agents, preservatives and the like.

The dose varies depending on the age, sex, body weight, degree of disease, type of composition of the present invention, dosage form, etc. of the patient to be administered. About 5 O mg should be administered. In addition, the above-mentioned daily dose can be administered once or divided into several times. When this compound is used as a food additive, the specific form of the food to which the additive is added includes, for example, powder, tablets, beverages, ingestible gel or syrup mixed liquid, For example, seasonings, Japanese confectionery, Western confectionery, frozen confectionery, beverages, spreads, pastes, pickles, canned bottles, processed meat products, fish 'fishery products, milk' processed eggs products, processed vegetables products, processed fruit products, processed cereal products, etc. You can list general food and drinks and tastes. It can also be added to feed and feed for domestic animals, poultry, bees, cormorants, fish and other domestic animals.

The dose varies depending on the age, sex, body weight, degree of disease, type of composition of the present invention, dosage form, etc. of the patient to be administered. 50 O mg may be administered. In addition, the above-mentioned daily dose can be administered once or divided into several times. Example

The following examples further illustrate the present invention. Example 1 The synthesis of this compound is described in Example 1_1. Example 1 1 1 Compound [Synthesis of Compound No. (16)]

A 2.5 ml solution of resorcinol 0.1 g and ethyl 4, 4, 5, 5, 5-pentafluoropropionyl acetate 0.23 g in trifluoroacetic acid was stirred for 15 hours under reflux. The mixture was stirred for 15 hours under reflux. The reaction solution was concentrated under reduced pressure, and the resulting residue was subjected to silica gel column chromatography and recrystallized from a mixed solvent of chloroform and hexane to give a white powder of compound No. (16), 4 Omg Got.

ιΗ— 匪 R (270MHz, DMSO- d ₆ ) δ (ppm): 6. 73 (s, 1H), 6. 83 (d, 1H, J -2.4 Hz), 6. 89 (dd, 1H, J = 2. 7 Hz, 8. 9 Hz), 7. 61 (d, 1 H, J = 8.4 Hz) Example 2 Measurement of the ability of this compound to inhibit type 3 17 / 3-hydroxysteroid dehydrogenase

(2-1) Preparation of plasmid for human type 3 173-hydroxysteride dehydrogenase expression

The cDNA of human type 3 176-hydroxysteride dehydrogenase gene was cloned from human testis cDNA (MTC Multiple Tissue cDNA Panels / Human II, Clontech) by PCR. Add a primer consisting of the base sequence shown in SEQ ID NO: 1 and a primer consisting of the base sequence shown in SEQ ID NO: 2 to a final concentration of 0.4 M, respectively, and use a heat-resistant DNA polymerase (TaKaRa LA Taq, Yukara Bio) The PCR reaction was then performed with a thermal cycler (GeneAmp PCR System 9700, Applied Biosystem). The reaction conditions are as follows: 95 ° C for 5 minutes, 95 30 seconds, then 59 ° C for 1 minute, 72 ° C for 45 seconds, 30 cycles, and 72 ° C for 7 minutes. Keep warm. The obtained PCR product was subjected to agarose gel electrophoresis, and about 1. lkb of DNA was recovered.

The expression vector pcDNA3.1 / Hyg (Invitrogen) was digested with the restriction enzyme Hindlll (Yu-Karabio), and then treated with T4 DNA P01 ymerase (DNA Blunting Kit, Yu-Kara Bio) to smooth the ends. The approximately 1.1 kb DNA obtained above and the linearized expression vector were mixed and ligated using T 4 DNA Ligase (DNA Ligation Kit Ver.2.1, Yukara Bio). It was introduced into a DH5 combi- tive cell (Yu-Kakara Bio). From the obtained transformant, a plasmid for human 3 type 17 / S-hydroxysteride dehydrogenase expression was obtained. The nucleotide sequence of the DNA cloned into the plasmid is the corresponding base sequence of the human type 3 1 β-hydroxysteride dehydrogenase gene disclosed in GenBank, a public database, under the registration number U05659. Matched the sequence.

(2-2) Preparation of transiently expressing cells of human type 17ι3-hydroxysteroid dehydrogenase

HeLa cells 1.6Xl0 ⁶ cells were seeded cell suspension of 1 OML was suspended in a cell culture dish with a diameter of 10cm in 10% FCS-containing D-MEM medium, and allowed to stand at 20-24 hours C0 ₂ incubator. Add the plasmid for human type 3 obtained in (2-1) 17) 3-hydroxysteroid dehydrogenase expression to FCS-free D-MEM medium 300 1 and gently pipet twice to mix gently. I let you. To the obtained DM solution, 501 transfection reagent (PolyFect Transfection Reagents Qiagen) was added, mixed by pipetting 5 times, and allowed to stand at room temperature for 10 minutes. To this DNA solution was added lmL of 10 CS])-MEM medium, and the DNA solution was mixed by pipetting twice. The medium of the cells after standing for 20 to 24 hours was replaced with 8 mL of fresh medium, and the MA solution was added. After standing at 20-24 hours C0 ₂ incubator, the cells were used for enzyme activity measurement plated on 96 © El plate detached with trypsin.

(2-3) Measurement of enzyme activity

Evaluation was performed by adding androstenedione to HeLa cells in which human type 17; 3-hydroxysteroid dehydrogenase was transiently expressed, and measuring the concentration of testosterone produced by the conversion. HeLa cells introduced with the human 3 type 17; 6-hydroxysteroid dehydrogenase expression plasmid prepared in (2-2) were suspended in D-MEM medium containing 10% FCS. The resulting cell suspension is lx cells per well on a 96 well plate (IOO D was added, diluted in 20-24 hours C0 ₂ incubator one was allowed to stand. After standing to take disconnect the medium, newly a FCS-free medium was added 80 L. 1% DMS0 containing FCS-free medium 10 L of the compound solution was added and left still in a C0 ₂ incubator for 30 minutes, and 10 siL of androstenedine solution diluted to 5 OOnM in a FCS-free medium was added to it for 20 minutes. and allowed to stand at ₂ incubator. Thereafter, time-resolved fluorescence reagent (DELFIA Testo sterone reagents, Perkin Elmer I) was used to measure the concentration of testosterone in the medium in accordance with instructions of the reagent comes. measurements multi functional · Plate reader (manufactured by Tecan, Ultra) was used Measurement wavelength was excitation 340ηιη, fluorescence was 612nm, Lag time was 400 sec, Integration time was 400 sec. After adding 10 L each of 1IDMS0-containing FCS-free medium and 500iiM androstenedione solution, the testosterone concentration was measured in the same way, and as a control value for 100% inhibition, the above-mentioned 20 ˜24 hours after addition of lO ^ L to 1¾D MS0-containing FCS-free medium and FCS-free medium, the testosterone concentration was measured in the same manner. The inhibition rate of 173-hydroxysteride dehydrogenase activity by each concentration of compound was calculated Compound (A) represented by Compound No. (1) is a type 3 17 i3-hydroxyloxylo at 5 / M concentration. The enzyme inhibition rate of idedehydrogenase was 100% Compound Nos. (2) to (11), (14), (16), (18) to (20), (22), (23) The compound (A) represented by any of the above is human type 3 173-H The enzyme inhibition rate of loxosteroid dehydrogenase was 80% or more Compound (A) represented by Compound No. (1 3) or (15) is human type 3 17 / 3-hydroxyloxy at a concentration of 1 The enzyme inhibition rate of dodehydrogenase was over 70%. The compound (Α) represented by the compound number (17) had an enzyme inhibition rate of human type 3 17ι8-hydroxycysteide dehydrogenase of 70% or more at a concentration of 1Ο. Industrial applicability

The present invention inhibits the activity of type 3 17 3-hydroxysteroide dehydrogenase and suppresses the enhancement of male hormone activity in tissues, thereby It is possible to develop and provide a composition or the like for treating or preventing a human dependent disease. Sequence listing free text

SEQ ID NO: 1

An oligonucleotide primer designed to amplify the human type 1 7 3-hydroxysteride dehydrogenase gene

SEQ ID NO: 2

An oligonucleotide primer designed to amplify the human type 3 1 7] 3-hydroxysteride dehydrogenase gene

Claims

14 Claim

1. Formula (I)

[Where

X, represents a hydrogen atom, a methoxymethyl group, a phenyl group, a benzyl group or a C 卜 C4 alkyl group which may be substituted with a fluorine atom,

Y x represents a hydrogen atom, a halogen atom, a C 卜 C4 alkyl group, a methoxymethyl group or a phenyl group,

Ζ, represents a hydrogen atom or a halogen atom,

R _r represents a hydrogen atom, a CI-C4 alkylcarbonyl group or a CI-C4 alkylaminocarbonyl group.

However, X! When is a hydrogen atom or a methyl group and _Yr is a hydrogen atom, cannot be a hydrogen atom. ]

A composition for inhibiting type 3 17 β-hydroxysteride dehydrogenase, which comprises a coumarin compound represented by the formula (1) and an inert carrier.

2. A coumarin compound represented by the formula (II).

3. A type 3 17) 3-hydroxysteroid dehydrogenase inhibitor composition comprising the coumarin compound according to claim 2 and an inert carrier.

4. It is contained as an active ingredient for inhibiting type 3 17 / 3-hydroxysteride dehydrogenase in the composition of type 3 1 3; 6 to hydroxysteroide dehydrogenase inhibitor according to claim 1. Use of coumarin compounds.

5.3 Type 3 1 7) Use of a coumarin compound according to claim 2 as an active ingredient for inhibiting 3-hydroxysteride dehydrogenase.

6. A method of inhibiting type 3 1 7) 3-hydroxysteroid dehydrogenase, wherein the method comprises a therapeutically effective amount of type 3 17 / 3-hydroxysterodedehydrogenase inhibition according to claim 1. A method comprising administering a coumarin compound contained as an active ingredient in a composition to a patient in need of such inhibition.

7. A method of treating or preventing an androgen-dependent disease, said method comprising a therapeutically effective amount of type 3 1 7 according to claim 1 to a patient in need of said treatment or prevention. ) A method comprising administering a coumarin compound contained as an active ingredient in a 3-hydroxysteroid dehydrogenase inhibitor composition.

8. Male hormone-dependent diseases include prostate cancer, benign prostatic hyperplasia, prostatic intraepithelial neoplasia, hirsutism, acne, seborrhea, androgenetic alopecia, sexual prematurity, adrenal hypertrophy or polycyst The method according to claim 7, which is ovarian ovary syndrome.