WO2011047596A1

WO2011047596A1 - Use of hydroxy benzopyrone compounds for preparing medicine useful for for precaution and treatment of fracture and osteoporosis

Info

Publication number: WO2011047596A1
Application number: PCT/CN2010/077337
Authority: WO
Inventors: 李靖; 孟坤
Original assignee: 北京盛诺基医药科技有限公司
Priority date: 2009-10-20
Filing date: 2010-09-26
Publication date: 2011-04-28
Also published as: CN102038674A

Abstract

Disclosed is a use of compound of formula(Ⅰ), pharmaceutically acceptable salts or pharmaceutical composition thereof for preparing a medicine for the precaution and the treatment of fracture, osteoporosis and the disorders regulated by female hormone receptor subtypes of ER-α and ER-β.

Description

Description Hydroxybenzopyrone compounds are prepared

Application in the prevention and treatment of fractures and osteoporosis drugs

This invention relates to the use of hydroxybenzopyrone compounds. The compounds of the present invention are useful for preventing bone fractures, reducing the incidence of fractures, and promoting the repair of damaged bones after fractures, as well as preventing and treating osteoporosis. The invention further relates to pharmaceutical compositions comprising these compounds, and to the use of said compounds, alone or in combination with other agents, to prevent fractures and to promote repair of damaged bones after fracture, and to prevent and treat osteoporosis.

Background technique

Estrogen is a steroid hormone produced by the endocrine system in the body and plays an important role in the reproductive system, cardiovascular system, central nervous system, immune system and skeletal system. Recent studies have shown that estrogen exerts biological effects through two signaling pathways, the nuclear pathway and the cell membrane pathway; estrogen and its receptor signal transduction system are widely involved in cell growth, differentiation and apoptosis in multiple system tissues in vivo. . A large number of studies have confirmed the presence of estrogen receptors in the skeletal system, and their receptor expression levels are associated with skeletal development and osteoporosis; ER-α-mediated signal transduction pathways are likely to be involved in osteoporosis The formation and development process.

Medical research has long established that serum estrogen concentrations below a certain level will result in rapid, sustained bone loss and osteoporosis. Estrogen has been used to treat osteoporosis for many years. Clinical observations have found that estrogen has a positive effect on reducing rapid bone loss after menopause, increasing bone mass, alleviating bone pain caused by osteoporosis, and reducing the incidence of fracture. Numerous studies have also confirmed that estrogen receptors are present in the skeletal system, and their receptor expression levels are associated with skeletal development and osteoporosis; estrogen receptor-mediated signal transduction pathways are involved in the formation of osteoporosis and development process. Selective estrogen receptor modulators developed for estrogen receptors, such as raloxifene, exhibit potent estrogen stimulatory effects on the skeletal system, increase bone density, and improve osteoporosis.

Current studies have shown that estrogen receptors are divided into two subtypes, alpha and beta; wherein ER-ct includes three homologs, ER-a66, ER-a46 and ER-a36. The isoform ER-a36 is from the first ER-a66 gene The promoter in the intron starts transcription and is encoded by ER- (a partial exon of x66. Therefore, ER-CL36 lacks two transcriptional functional regions, retaining the DNA-binding functional region and the dimerization functional region; It is important that the hormone ligand binding region of ER-a36 lacks a partial helix region, thus completely changing its specificity and affinity for binding to hormone ligands. ER-CL36 is mainly distributed in cell membranes and cytoplasm, and is present in small amounts in In the nucleus, ER-CL36 regulates cell growth and apoptosis through the estrogen cell membrane signal transduction pathway (1. ZY,

Wang; XT, Zhang; P, Shen; BW, Loggie; YC, Chang; TF, Deuel. A variant of estrogen-α, hER-a36: Transduction of estrogen- and antiestrogen-dependent membrane-initiated mitogenic signaling. 2006, 103(24): 9063-9068. 2. L, Lee; J, Cao; H, Deng; P, Chen; Z, Gatalica; ZY, Wang. ER-a36, a novel variant of ER-a, is Expressed in ER-positive and -negative human breast carcinomas. Anticancer Res.

2008, 28(lB): 479-483. )

Summary of the invention

The compounds of the present invention are useful as modulators of ER-ct and ER-β bioinformatics systems for the prevention and treatment of fractures and osteoporosis.

The present invention relates to a compound of formula (I), and a pharmaceutically acceptable salt or pharmaceutical composition thereof for the preparation of a prophylactic, therapeutic fracture and bone

RR ² and R ³ are independently of each other selected from the group consisting of hydrogen, methyl, ethyl, propyl, and isopropyl.

In a particularly preferred embodiment of the invention, the compound of formula (I) is: 3,5,7-trihydroxy-2-(4-methoxyphenyl)-8-(3-methyl-2- Butenyl-)-4H-benzopyran-4-one (aka: Epimedin, Icaritin), that is, the compound of formula (I) has the structure of formula (Π):

(Π)

In a particularly preferred embodiment of the invention, the compound of formula (I) is a demethylated product of a compound of formula (Π): 3,5,7-trihydroxy-2-(4-hydroxyphenyl 8-(3) -Methyl-2-butene-)-4H-benzopyran-4-one (aka: demethylated icariin),

(III)

The compound of the formula (I), and a pharmaceutically acceptable salt or pharmaceutical composition thereof, are prepared by the estrogen receptor ER-α and ER-β subtypes

among them,

R1, R2 and R3 are independently of each other selected from the group consisting of hydrogen, methyl, ethyl, propyl and isopropyl.

In a particularly preferred embodiment of the invention, the compound of formula (I) is: 3,5,7-trihydroxy-2-(4-methoxyphenyl)-8-(3-methyl-2- Butenyl-)-4H-benzopyran-4-one (aka: Epimedhasin, lcaritin), that is, the compound of formula (I) has the structure of formula (Π):

(Π)

In a particularly preferred embodiment of the invention, the compound of formula (I) is a demethylated product of a compound of formula (Π): 3,5,7-trihydroxy-2-(4-hydroxyphenyl 8-(3) -Methyl-2-butene-)-4H-benzopyran-4-one (aka: demethylated icariin), that is, the compound of formula I) has the structure of formula (ΠΙ):

(III)

The compound of the formula ω, (π), cm) and a pharmaceutically acceptable salt or pharmaceutical composition thereof, for use in the preparation of a medicament for the regulation of estrogen receptor ER-α and ER-β subtypes For use, the ER-ct subtypes include ER-a36, ER-a46 and ER-a66.

The compounds of the formula (I), (Π), (III) and pharmaceutically acceptable salts or pharmaceutical compositions thereof are modulators of the estrogen receptor ER-α and ER-β subtypes.

The term "compounds of formula (I)" includes compounds of the formulae as defined herein, as well as all embodiments, preferred embodiments, more preferred embodiments and particularly preferred embodiments of the compounds, including the compounds disclosed in the specific names or examples, Each of these compounds is a particularly preferred embodiment of a compound defined by the formula. The term "compound of the present invention" means any compound of the formula (I) as defined above.

The invention also relates to salts, polymorphs, solvates and hydrates of the compounds of the invention. The term "pharmaceutically acceptable salt" as used herein, unless otherwise stated, includes salts which may be present in the compounds of the invention.

The verb "treatment" as used in the present invention, unless otherwise stated, refers to the process of reversing, alleviating, inhibiting a disease or condition, or preventing such a disease or condition. The term "treatment" as used herein refers to the act of treatment, The treatment therein is as defined above.

The invention further relates to a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable carrier. For example, the pharmaceutical composition may be administered orally in the form of a tablet, a capsule, a pill, a powder, a sustained release, a solution, or a suspension; the pharmaceutical composition may also be administered parenterally in the form of a sterile solution, suspension or emulsion. The pharmaceutical composition may also be administered topically in the form of an ointment or cream; the pharmaceutical composition may also be administered rectally in the form of a suppository. The pharmaceutical compositions may be presented in unit dosage form suitable for single administration of precise dosages. The pharmaceutical composition includes a conventional pharmaceutical carrier or excipient and a compound of the present invention as an active ingredient. In addition, the pharmaceutical composition may include other medical or pharmaceutical preparations, carriers, adjuvants. Suitable pharmaceutical carriers include inert diluents or fillers, water, and various organic solvents. The pharmaceutical composition of the present invention may contain other ingredients such as a flavoring agent, a binder, an excipient, etc., if necessary. Thus for oral administration, the tablets may contain various excipients, for example citric acid and various disintegrating agents such as starch, alginic acid and certain complex silicates and binders such as sucrose, gelatin and arabic may be used. gum. In addition, the tablets may also contain lubricants such as magnesium stearate, sodium lauryl sulfate and talc, which are often used in tablets. A similar type of solid composition can also be used with soft and hard filled capsules. Preferred materials include lactose or milk sugar and high molecular weight polyethylene glycol. When it is desired to administer the active compound of the invention orally, the aqueous suspension or elixirs can be combined with various sweetening or flavoring agents, pigments or dyes and, if desired, with emulsifiers or suspending agents, and diluents such as water, ethanol, Propylene glycol, glycerol or a combination thereof is administered in combination.

Exemplary parenteral administration forms include solutions or suspensions of the active compounds in sterile solutions, for example, propylene glycol or aqueous dextrose. Such dosage forms can be suitably buffered if desired. The aqueous composition of the present invention may include other pharmaceutically acceptable solutes, including additives and other drugs, as the case may be. Suitable additives are those well known in the art including, but not limited to, antioxidants, antibacterials, surfactants, chelating agents, sugars, and preservatives. The aqueous composition of the present invention can be administered by injection, wherein the injection can be intramuscular, intravenous or subcutaneous.

The compounds of the invention may be administered alone in terms of the intended route of administration and standard pharmaceutical practice, but are often administered with a suitable pharmaceutical excipient, diluent or carrier well known in the art. If appropriate, adjuvants may be added, including adjuvants, antioxidants, flavoring agents, or coloring agents. The compounds of the invention may be formulated into suitable pharmaceutical compositions to provide direct, delayed, modified, sustained, pulsed or controlled release, depending on the particular route of administration and the specificity of the release profile, and Treatment needs to correspond.

The compounds of the invention may be administered in a route such as, but not limited to, oral (including buccal, sublingual, etc.) forms well known in the art. In this case, "oral" refers to an oral administration method in which the oral use amount of the animal is expressly specified, for example, in a food or beverage, directly into the oral cavity, or freely selected. In the present invention, the term "animal" refers to warm-blooded animals having adaptive functions in the animal kingdom, such as dogs and humans.

Typical oral solid forms can include tablets, powders, multiparticulate formulations (granules), capsules, chewables, lozenges, films, patches, and the like. Typical oral liquid (including semi-solid and gelatinous) forms can include solutions, elixirs, gels, sprays, liquid-filled chews, and the like. Other oral administration forms may also be employed in which the active agent is suspended in a liquid or semi-solid carrier phase, such as a suspension.

The compounds of the invention may also be administered parenterally. The term parenteral administration means that the route of administration does not pass through the mouth. Preferably, for the compounds of the invention, parenteral administration may include topical and transdermal, rectal, vaginal, nasal, inhalation and parenteral administration (requiring means of penetration through the skin barrier by needle and needle-free methods, including implants) And reservoir).

Compositions of the invention include tablets. In a preferred embodiment, the tablets are prepared by a process selected from the group consisting of direct compression or wet, dry or melt granulation, melt coagulation processes and extrusion. In another embodiment, the tablet core portion of the compositions of the present invention may be single or multi-layered, coated with a suitable coating material well known in the art.

The oral liquid form of the compound of the present invention is preferably a solution or suspension in which the active compound is sufficiently dissolved or partially dissolved. In one embodiment, the solution comprises the active compound and a pharmaceutically acceptable solvent suitable for oral administration. In a preferred embodiment, the solvent is a solvent which exhibits good solubility for the compounds of the present invention. These solvents typically comprise a major portion of the formulation, i.e., greater than 50% by weight, preferably greater than 80%, such as 95%. In a preferred embodiment, the solution further comprises an adjuvant or an additive. In a preferred embodiment thereof, the additive or adjuvant is a taste masking agent, a palatant, a flavoring agent, an antioxidant, a stabilizer, a texture modifier, a viscosity modifier, and a solubilizer.

Another embodiment is a process for the preparation of a preferred oral liquid form of a compound of the invention (see the pharmaceutical composition portion) wherein, in a suitable temperature range, each preferred component is optionally mechanically agitated or in a manner which facilitates dissolution rate Mix by ultrasonic agitation.

The preparation of the compound of the present invention is isolated and extracted from the raw material of the Chinese medicinal material. Preparation of the compounds of the invention According to the method described in another invention patent "Preparation method of icariin" (Meng Kun, CN 101302548A), the chemical ingredient icariin is isolated and extracted from the Chinese medicinal material Epimedium, that is, the embodiment of the present invention 1 Compound (11).

Demethylated icariin was purchased from Shanghai Yousi Biotechnology Co., Ltd.

The 24-week-old SD rats were treated with sham operation (Sham) or ovariectomy (OVX), and were randomly divided into groups of 8 animals each. Rats received different drug interventions. After 8 weeks, the uterus of each animal was collected and the uterine weight was weighed. The femurs of each animal were collected and the femur bone mineral density (Femur BMD) was measured. The expression of ER-CL36 receptor in rat endometrium was determined by immunohistochemistry. Western blot was used to detect ER-a66 and ER-a36 in rat endometrium and distal femoral cancellous bone. Receptor protein expression. DRAWINGS

Figure 1. Effect of compound (Π) on femur bone density and uterine weight in rats.

1, sham operation group; 2, ovariectomy group; 3, estradiol group; 4, compound (Π) group; 5, raloxifene group. #P<0.05, compared with the sham operation group; *P<0.05, compared with the ovariectomy group.

Figure 2. Effect of compound (Π) on endometrial ER-CL36 receptor expression.

A is a sham operation group, B is an oophorectomy group, C is an estradiol group, D is a raloxifene group, and E is a compound (Π) group.

Figure 3. Effect of compound (Π) on endometrial ER-CL36 receptor protein expression.

Figure 4. Effect of compound (Π) on the expression of ER-CL36 receptor protein in cancellous bone of distal femur.

Among them, Fig. 3 and Fig. 4 are 1 : sham operation group; 2: oophorectomy group; 3: raloxifene group; 4: estradiol group, 5: compound (Π) group. Specific implementation method

The following examples illustrate the preparation of the compounds of the invention. It should be understood that the present invention is not limited by the specific details provided below. Experimental material 1

(Π)

3,5,7-trihydroxy-2-(4-methoxyphenyl)-8-(3-methyl-2-butenyl)-4Η-benzopyran-4-one:

The preparation of the compound of this example is carried out according to the method described in another invention patent "Preparation method of icariin" (Meng Kun, CN 101302548A), and the chemical monomer icariin is isolated and extracted from the Chinese medicinal material Epimedium. That is, the compound of the examples of the present invention.

Experimental material 2

(III)

3,5,7-trihydroxy-2-(4-hydroxyphenyl)-8-(3-methyl-2-butene-)-4H-benzopyran-4-one:

This example compound was purchased from Shanghai Yousi Biotechnology Co., Ltd.

Experimental Example 1 Biological experiment of the compound of formula (I)

It has been previously described that the present invention relates to a compound of formula (I), and a pharmaceutically acceptable salt or pharmaceutical composition thereof, for use in the prevention and treatment of fractures and osteoporosis. The compound of the example (Π) is now taken as an example and will be described in detail.

(1) The specific experimental method of this experiment is as described above. The experiment was divided into 5 groups, namely sham operation group, ovariectomy group, estradiol group, compound (Π) group, raloxifene group; compound (Π) was used at a dose of 100 μ _§ /13⁄4.

(2) Experimental results: It was found through experiments that the femur density and uterine weight of the rats after ovariectomy were higher than that of the artificial hand. The rats in the experimental group were significantly reduced, and the compound (Π) significantly increased the bone density of the femur in rats, while the compound

(Π) had no significant effect on rat uterine weight (Fig. 1). Immunohistochemistry and Western blot analysis showed that ER-CL36 protein was positive in endometrial glands of rats in sham operation group and ovariectomy group; ER-CL36 protein in endometrial gland of rats in raloxifene group was weakly expressed; The ER-a36 protein was negatively expressed in the endometrial glands of the alcohol group and the compound (Π) group (Fig. 2 and Fig. 3). The cancellous bone tissues of the distal femur of each group were extracted. Western blot analysis showed that the expression of ER-a36 receptor protein in ovariectomized rats was significantly higher than that in other groups. The ER-C136 receptor in estradiol rats The protein expression was less than that of the sham-operated group, and the expression of ER-CL36 receptor protein was significantly reduced in the raloxifene group and the compound (Π) group (Fig. 4).

Claims

Claim

A compound of the formula α) and a pharmaceutically acceptable salt or pharmaceutical composition thereof for the preparation of a medicament for the prevention, treatment of fractures and osteoporosis

(I)

among them

2. The use according to claim 1, wherein in the compound of the formula (I), R ¹ is a methyl group, and R ² and R ³ are each hydrogen, that is, a compound having the formula (Π structure:

(Π)

3. The use according to claim 1, wherein R^R ² and R ³ in the compound of formula (I) are both hydrogen, ie, having the structure of formula (III)

4. The use of a compound of formula (I), and a pharmaceutically acceptable salt or pharmaceutical composition thereof, for the manufacture of a medicament for a disease associated with estrogen receptor ER-α and ER-β subtypes,

(I)

among them

5. The use according to claim 4, wherein R ¹ in the compound of formula (I) is methyl, R ² and R ³ are all hydrogen, ie have the formula

6. The use according to claim 4, wherein R^R ² and R ³ in the compound of formula (I) are both hydrogen, ie, having the structure of formula (III)

(III)

7. The use according to any of claims 4-6, said ER-α subtype comprising ER-a36, ER-a46 Wokou ER-a66.

The use according to any one of claims 1 to 3, wherein the compound of the formula (I) and a pharmaceutically acceptable salt or pharmaceutical composition thereof are modulators of the ER-ct and ER-β subtypes of the estrogen receptor .