CN101284795B - Non-steroidal male hormone receptor regulating agent and medical uses thereof - Google Patents

Abstract

The invention relates to a non-steroidal male sex hormone receptor modulator or the receivable slat thereof in pharmacy, in particular to a compound for preparing non-steroidal drugs used for preventing and/or curing the partial androgen deficiency in aging males, osteoporosis, myophagism, female sexual insufficiency, anorexia, female polytrichia, severe androgen-dependent type alopecia, acne, cachexia, benign prostatic hyperplasia, prostatic cancer, breast cancer, alzheimer disease, parkinsonism syndrome, etc. The invention also relates to a drug combination containing a general formula I compound or an acceptable salt thereof in pharmacy, and the use of the drug combination containing the general formula I compound or the acceptable salt in pharmacy for preparing drugs to cure the abovesymptoms or diseases.

Description

Class I non-steroid androgen acceptor regulator and medical usage thereof

Technical field

The present invention relates to class I non-steroid androgen acceptor regulator and pharmaceutical composition thereof and preventing and/or treating symptom that male sex hormone and androgen receptor functional disorder cause or the purposes in the disease medicament; Include, but are not limited to that the middle-aging male male sex hormone lacks partly that syndrome, osteoporosis, muscle wasting (Sarcopenia), femal sexual function are complete, apocleisis, female hirsutism, the alopecia of severe male sex hormone dependent form, acne, evil matter disease, benign prostatic hyperplasia, prostate cancer, mammary cancer, alzheimer's disease (Alzheimer ' s disease; AD) and parkinsonism (Parkinson ' s disease, PD) etc.

Background technology

Male sex hormone (Androgen) is one type of important steroidal class sexual hormoue in the human body; Through with androgen receptor (Androgen Receptor; AR) combine to promote cytodifferentiation and tissue growth, participate in numerous crucial physiological functions such as male fetus reproductive organ (like prostate gland, seminal vesicle, epididymis etc.) formation, secondal sexual character growth with keep and sperm generation etc.All have a certain proportion of male sex hormone in the sex body, as androsterone (Testosterone, T), (Dihydrotestosterone is DHT) with adrenosterone etc. for androstanediol.In female body, androsterone can change oestrogenic hormon in most of target organ, but the physiological function of in portion of tissue such as brain, bringing into normal play.In prostate gland and skin, androsterone is converted into DHT by 5, and compares with T, and DHT will exceed 3-5 times to the avidity of AR.

AR is a member of nuclear receptor superfamily; Metal ligand activated transcription factor; Be distributed widely in propagation and non-propagation tissue; Comprise prostate gland and seminal vesicle, male and female genital organ, testis, ovary, skin, hair follicle, sweat gland, cartilage, cardiac muscle, bone and unstriated muscle, adrenal cortex, liver, pineal gland and a plurality of palliumes district and cortex inferior segment, comprise dynamoneure (Negro-Vilar, J Clinical Endocrinology and Metabolism; 1999,84:3459-3462).AR albumen has three structural domains: N end structure territory (N terminal domain; NTD), DNA combines territory (DNA binding domain; DBD) and ligand binding domain (Ligandbinding domain, LBD) (He B, Kemppainen JA; 1999, J.Biol.Chem.274:37219-25).After the LBD of male sex hormone and AR formed mixture, (Androgen response element ARE) combined, and plays the effect that activates or suppress expression of target gene, thereby regulates and control the physiological function of target tissue to reply original paper with the male sex hormone that is positioned at the target gene promoters district.The generation of a lot of AR relative diseases, or because hormonal readiness is not normal, or, destroyed normal interaction equilibrium relationship because function of receptors is disorderly.In diseased individuals, medicine can reach the effect of treating relative disease through strengthening or suppressing the receptor activation level.Therefore, in the middle of the treatment multiple disease and symptom relevant with androgenic effect, AR is important target.For metabolism and incretion disease; The AR regulator can be used for preventing/treating and age and male hormone metabolism diseases associated, as the elderly men male sex hormone reduce that disease, women's hypoandrogenism disease, osteoporosis, muscle wasting, femal sexual function are complete, evil matter disease when apocleisis, hirsutism, the alopecia of severe male sex hormone dependent form, acne, several morbid state etc.In oncology, the AR regulator can be used for treating benign prostatic hyperplasia, hormonal dependent prostate cancer and mammary cancer etc.; For nervous system disorders, the AR regulator can be used for treating melancholia, presenile dementia and parkinsonism etc. (Segal S., Naraynan R.and Dalton J.Expert Opin.Investig.Drugs2006,15:377-387).

Clinical AR regulator commonly used is divided into steroidal class and nonsteroidal from structure.Steroidal class male sex hormone in body natural generation; The ester (like cipionate, propionic ester, phenylpropionic acid ester, cyclopentanepropanoiacid acid ester, heptanoate and decylate) that also comprises androsterone, and other synthetic type male sex hormone (like 7-methylnortestosterone and acetic ester thereof).Then (Cyproterone acetate CPA) is representative to androgen antagonist with cyproterone acetate.Nonsteroidal AR regulator is used more have flutamide (Flutamide), RU-23908 (Nilutamide) and bicalutamide, and (Bicalutamide Casodex) etc., is receptor antagonist.Document and patent report have also the been arranged AR agonist of some non-steroidals, but all do not get into clinical application at present.Be that steroidal and nonsteroidal AR regulator all have certain spinoff, comprise causing the estrogen and androgen metabolism disorder and making gynaecomastia and hepatotoxicity etc.Spinoffs such as hyperplasia of prostate/patients with prostate cancer people takes for a long time that flutamide or bicalutamide gastrointestinal discomfort can occur, feel sick, vomiting, insomnia, weak, headache, anxiety, blurred vision and hyposexuality; And " androgen antagonist go down syndromes " (Antiandrogen withdrawalsyndrome can appear in single taking behind certain androgen antagonist medicine; AWS); Rising rapidly, gross tumor volume increase the repressed PSA level of script again after showing as medication for some time; Can only withdrawal or migrate other androgen antagonist medicines (DickerAP, 2003, Lancet Oncol.4:30-36; Laufer M, Sinibaldi VJ, 1999, Urology54:745].There is report to show that there is interaction in the AR regulator with the antithrombotics tonka bean camphor in use in addition.Therefore, press for the nonsteroidal AR regulator that research and development has brand-new chemical structure clinically.

Researchist of the present invention discloses class I non-steroid androgen acceptor regulator and preparation method thereof, pharmaceutical composition and purposes in the CN200510026252.2 patented claim.Though in purposes, disclose the effect that this compounds has androgen receptor modifier, only disclose the purposes of compound, and its purposes as the androgen receptor agonist aspect is not done any elaboration as androgen receptor antagonists.

Summary of the invention

The defective that exists to above-mentioned prior art or not enough, the present invention is through chemosynthesis and structure activity study, prepares and optimized one type of novel nonsteroidal micromolecular compound.Androgen receptor competition combines experiment to show, this compounds to the avidity of acceptor less than 10nM, suitable with DHT; Luciferase reporter gene cotransfection experiments and prostate cancer cell strain LNCaP proliferation experiment show that it has excitement of AR part and antagonistic activity, has showed the potentiality that are developed further into to novel androgen receptor modifier.

Therefore, the object of the present invention is to provide one type of Nonsteroic androgen acceptor regulators compound or its pharmacy acceptable salt with following general formula I structure.

Another purpose of the present invention is to provide the compound that comprises the general formula I structure or the pharmaceutical composition of its pharmacy acceptable salt.

Compound or its pharmacy acceptable salt that a purpose more of the present invention is to provide the general formula I structure as the nonsteroidal androgen receptor modifier in the preparation prevention or/and symptom that treatment causes because of male sex hormone and androgen receptor functional disorder or the purposes in the disease.

Compound with general formula I structure of the present invention or its pharmacy acceptable salt as the nonsteroidal androgen receptor agonist in the preparation prevention or/and symptom that treatment causes because of male sex hormone and androgen receptor functional disorder or the purposes in the disease.

Compound with general formula I structure of the present invention or its pharmacy acceptable salt as the nonsteroidal androgen receptor antagonists in the preparation prevention or/and symptom that treatment causes because of male sex hormone and androgen receptor functional disorder or the purposes in the disease.

Nonsteroic androgen acceptor regulators provided by the invention or its pharmacy acceptable salt have the structure of following general formula I:

Wherein: R is Cl or Br; Specific examples is compound 3-phenyl-3-(4-oil of mirbane amido)-1-(4-chloro-phenyl-)-1-acetone (MWW6003) and 3-phenyl-3-(4-oil of mirbane amido)-1-(4-bromophenyl)-1-acetone (MWW6015), and its structural formula is following:

For compound or its pharmacy acceptable salt of general formula I structure of the present invention, when having chiral carbon in the molecule, it is raceme or optically active body.

The compound of general formula I structure of the present invention can obtain acceptable salt on its pharmacology through conventional salifiable method pharmaceutically with any suitable acid, and for example, described acid is mineral acid, example hydrochloric acid, Hydrogen bromide, nitric acid, sulfuric acid, phosphoric acid etc.; Organic acid is such as formic acid, acetate, propionic acid, phenylformic acid, toxilic acid, fumaric acid, succsinic acid, tartrate, Hydrocerol A etc.; Alkylsulphonic acid is like methylsulphonic acid, ethylsulfonic acid etc.; Aryl sulfonic acid all can use like Phenylsulfonic acid, tosic acid etc.

Pharmaceutical composition provided by the invention comprises compound or its pharmacy acceptable salt of one or more general formula I structures of treating significant quantity, and this pharmaceutical composition can further contain one or more pharmaceutically acceptable carrier or vehicle.

Its ideal ratio of pharmaceutical composition provided by the present invention is, compound of Formula I or its pharmacy acceptable salt account for gross weight than 50%～99.5% as activeconstituents, and rest part accounts for gross weight than below 50%.

A purpose more of the present invention provides through giving androgen receptor modifier as herein described or its pharmacy acceptable salt (comprising its pharmaceutical composition), prevent and/or treat the various symptoms that cause because of male sex hormone and androgen receptor functional disorder or the method for disease as androgen receptor agonist and/or antagonist, includes but not limited to that osteoporosis, muscle wasting, femal sexual function are complete, apocleisis, female hirsutism, the alopecia of severe male sex hormone dependent form, acne, stearrhea, evil matter sick (including but not limited to AIDS), benign prostatic hyperplasia, prostate cancer, androgenetic alopecia, the optimum or malignant tumour (for example occurring in the cancer of tissues such as mammary gland, brain, skin, ovary, bladder, lymph, liver, kidney and pancreas) of expression androgen receptor, endometriosis, polycystic ovary syndrome, Alzheimer, parkinsonism, androgen-dependent/age related disease (partly lacking the amyotrophy of syndrome, male or female sexual dysfunction and non-sickbed patients etc. like the middle-aging male male sex hormone).Be preferably applied to treatment/prevention androgen-dependent tumour, especially prostate cancer especially, reduce the sickness rate of prostate cancer, stop the generation of prostate cancer or cause prostate cancer to disappear.

Description of drawings

To be compound combine the vigor test with DHT to the competitiveness of AR to Fig. 1.Compound MWW6003 and MWW6015 are suitable to avidity and the DHT of AR, both IC ₅₀Value is respectively 2.23 and 3.48nM, the IC of DHT under the similarity condition ₅₀Be 7.75nM.

Fig. 2 is the cytotoxicity test of compound in the HeLa cell.After compound MWW6003 and MWW6015 and HeLa cell were hatched three days, with the influence of its cell growth of MTT method mensuration, both only showed low cytotoxicity, the wherein IC of compound MWW6003 ₅₀Value is 2.8 μ M, the IC of MWW6015 ₅₀Between 10～40 μ M.

Fig. 3 is the pharmacologically active test of compound in MDA-MB-453 cell reporter gene transfection experiment.Dihydrotestosterone (DHT) thus the expression of luciferase increases chemiluminescence readings in the inducing cell, compound shows as the antagonistic action of androgen receptor and suppresses the chemoluminescence of DHT inductive.MWW6003 and MWW6015 be display part AR agonist activity under activation pattern, and its usefulness is respectively 19.5% and 20.4% of DHT, EC ₅₀Be respectively 320.1nM and 159.6nM, the EC of DHT under the similarity condition ₅₀Be 0.6nM; Under the antagonism pattern, compound MWW6003 and MWW6015 have shown certain AR antagonistic activity, its IC ₅₀Be respectively 884.8nM and 958.1nM, and bicalutamide (Bicalutamide, IC BCA) under the similarity condition ₅₀Be 690.9nM.

Fig. 4 is the pharmacologically active test of compound in the LNCaP cell proliferation experiment.LNCaP is the hormonal dependent prostate cancer cell, and DHT and AR agonist all can be induced its propagation, and the AR antagonist can suppress DHT inductive cell propagation effect.The result shows that compound MWW6003 and MWW6015 all show certain activity trend under exciting and antagonism pattern.

Embodiment

In order to illustrate content of the present invention and not limited to by it, the present invention is divided into following trifle is described in detail.

Definition

Only if definition is arranged in addition, technology that the present invention is used and scientific term have same meaning with the general understanding of the current techique in field under the present invention.The application of all patents that derive from gene pool and other DBs that this place mentions, application, announcement and other publications and sequence are taken in comprehensively to be quoted as a reference.If the definition that the application of all patents that derive from gene pool and other DBs of definition that this joint is illustrated and this patent ginseng usefulness, application, announcement and other publications and sequence are taken in and quoted is set forth opposite; Or when inconsistent, the definition of illustrating with this joint is as the criterion.

Used herein, " one " or " one " refers to " at least one " or " one or more ".

Used herein; " regulator " refers to that one type can strengthen (like agonist; Have agonist activity) or suppress (like antagonist; Have antagonistic activity) biological activity of target protein (comprising enzyme, acceptor etc.) or the compound of function, and its strengthen only temporarily act on specific cells with restraining effect or organize in the generation of a certain particular event, like signal transduction, transcription activating etc.

Used herein; " the middle-aging male male sex hormone partly lacks syndrome " (Partial androgendeficiency of the aging male; PADAM) refer to the progressive minimizing that middle age back male sex's male sex hormone produces, clinical manifestation comprises fatigue, depression, sexual desire reduction, sexual dysfunction, erective dysfunction, hypogonadism, osteoporosis, alopecia, obesity, senile amyotrophy, osteopenia, benign prostatic hyperplasia, anaemia, mood and cognitive change and prostate cancer etc.The generation of PADAM is relevant with the male sex hormone environment also can be corrected through control male sex hormone environment.

Used herein, " osteoporosis " (Osteoporosis is a kind of skeletal diseases of general OP), it is characterized in that bone density is hanged down and the degeneration of osseous tissue, has increased the friability of bone and the susceptibility of fracture.Fracture of femur is result the most serious in the osteoporosis, has the patient of 5-20% dead in 1 year, and loses viability above 50% survivor.The elderly suffers from the dangerous maximum of osteoporosis, and its sickness rate significantly raises with aging population.In addition, the women has more the tendency of suffering from osteoporosis than the male sex.Male sex hormone and estrogenic physiological concentration play an important role to keeping the bone homeostasis in whole life.Therefore, when male sex hormone or oestrogenic hormon were lost, its result had increased the bony remodeling ratio, has made the balance that absorbs and form tend to absorb again, has promoted the total loss of bone amount.Suitably replenish male sex hormone and male sex hormone appearance material and help to increase bone density, improve osteoporosis, reduce the generation of fracture.

Used herein, " hyperplasia of prostate " refers to one type because of prostatic optimum adenomatoid hyperplasia around the urethra causes in various degree bladder outflow obstruction property disease or symptom, is also referred to as " benign prostatauxe ".Hyperplasia of prostate is one of modal disease of Urology Surgery, has become " the stealthy killer " that threaten men's health.Clinical statistics shows, the male sex during 40-79 year, after the sickness rate of benign prostate hyperplasia is about 50%, 80 years old then up to 80%.Along with rhythm of life is constantly accelerated, Benign Prostatic Hypertrophy day by day increases, and is rejuvenation trend.Benign prostatic hyperplasia also is prone to cause multiple potential complication, like AUR, urinary tract infection, gross hematuria, vesical diverticulum, calculus, uronephrosis and renal failure etc. in puzzlement patient daily life.Research shows that the intravital dihydrotestosterone of patient is the main inducement of benign prostatic hyperplasia, and the androgen antagonist treatment helps to improve its symptom.

Used herein, " prostate cancer " refers to one type of male reproductive system common malignancy, is main with gland cancer.Prostate cancer is a kind of serious male sex's geriatric disease, and is very high at the M & M of European and American areas, accounts for the first place ([Landis SH, Murray T, 1998, CA Cancer J.Clin.48:6-29] of male sex's malignant tumour.Though the sickness rate of China's prostate cancer is lower than American-European countries, in recent years along with the acceleration of aging population degree, the change of traditional food structure, and to the raising of this type medical diagnosis on disease level, sickness rate is remarkable growth situation.Can excision except that early prostate cancer, anti androgenic therapy be clinical preferred option.

Used herein; " hirsutism " refers to the crinosity symptom that one type of disease that increases because of 3male hormone secretion causes in the women; Many not only long not only thick but also black hairs have promptly been grown at the position that should not become mildewed; Perhaps hair is android type and distributes, the phenomenon of thick and heavy black, pubes ventrad of eyebrow even umbilical region development.

Used herein, " alopecia of severe male sex hormone dependent form " refers to one type of serious seborrheic alopecia, claims male pattern alopecia again.

Used herein, " acne " refers to one type of pilosebaceous chronic inflammatory diseases, and good sending out in face, thoracic dorsal shows as infringements such as acne, papule, warts, tubercle, tumour, claims acne of youth again.

Used herein; " Alzheimer " is one group of primary degeneration brain degenerative disease that the cause of disease is not bright, and pathological change is mainly the cortex diffuse atrophy, and ditch returns broadening; The ventricles of the brain enlarge; Neurone reduces in a large number, and it is thus clear that pathologies such as senile plaque, NFT, graininess cavity corpusculum, choline acetylase and acetyl choline content significantly reduce.The a lot of diseases of alzheimer's disease are in the senium, and latent disease is slowly irreversibly made progress (2 years or longer), is main with the intelligence infringement.Onset 65 years old with the former (pre-aged), have with sick family history more, pathological development is very fast, temporo page or leaf and top page or leaf pathology are more remarkable, and aphasia and appraxia are often arranged.In the experiment of one group of placebo; The cognitive disorder of patients with Alzheimer disease is improved after accepting androgen replacement therapy; Pointed out clinical value (Cherrier MM, Matsumoto AM, the Amory JK et al.2005 of androgen receptor in this disease; Neurology, 64:2063-2068).

Used herein, " Parkinson's disease " are central nervous system degenerative diseases common among the elderly, mainly show as patient motion slowly, other part of trick or health trembles, health loses flexibility, muscle rigidity.Parkinson's disease are only second to tumour and cardiovascular and cerebrovascular diseases to the harm of patient's viability, thereby are called as the elderly's " the 3rd killer ", " chronic cancer ".People such as Okun confirm male sex hormone be supplemented with help improve this sick symptom (Okun MS, Walter BL, Mcdonald WM, et al.2002, Arch.Neurol.59:1750-1753).

" significant quantity " that is used to treat the compound of a certain specified disease used herein refers to enough improve or alleviate to a certain extent the amount of the sick symptom that accompanies therewith.This dosage can the single dose administration, also can be according to the regimen administration.But this dosage cure diseases, but be typically administration in order to improve this symptom.Possibly need for improving the symptom repeat administration.

Used herein, " acceptable salt, ester or other verivates on the pharmacology " comprises any salt, ester or verivate that those skilled in the art are easy to prepare with currently known methods.The compound of deriving like this and generating can not have toxic action to animal and human's administration.This compound or have pharmaceutical activity, or prodrug.

Used herein, " treatment " refers to that disease and symptom are improved with any way, or other useful changes.Treatment also comprises the application of The compounds of this invention on medicine.

Used herein, the symptom that gives a certain specified disease of a certain specific medication compsn " improvement " is meant any alleviating, and is no matter permanent, interim, over a long time, of short duration, can both owing to or relevant with using of this pharmaceutical composition.

Used herein; " pure basically " is meant enough even; Can not survey impurity for estimating the standard method of analysis that purity uses by one of skill in the art, said standard method of analysis is just like thin layer chromatography (TLC), gel electrophoresis and HPLC (HPLC).Even perhaps enough pure also refer to be further purified can not change the observable physicochemical property of this material, for example enzymic activity and biological activity.Being used for purifying compounds and making chemical pure basically method, is known in those skilled in the art.Yet chemical pure basically compound can be the mixture of steric isomer or isomers.In this case, be further purified the specific activity that perhaps can increase compound.

Used herein, " prodrug " is meant a kind of compound of vivo medicine-feeding, and this compound can be by metabolism, or be converted into biologically, on the pharmacology or the activity form on the therapeutics.In order to make prodrug, pharmaceutical active compounds will be modified, and this active compound is produced through metabolic process again.Prodrug can be designed to change its metabolic stability, or the precursor of transportation characterization, to cover its spinoff or toxicity, improves the sense of taste of medicine, or changes other characteristics.Rely on the knowledge of pharmacokinetics and medicine internal metabolism, in case active compound is known on the pharmacology, those skilled in the art just can design the prodrug of this compound.[referring to Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, 1985, pages388-392].

Term " basically " is identical even or similar, can in context, change to some extent the understanding of correlation technique according to those skilled in the art, and be generally at least 70%, is preferably at least 80%, more excellently be at least 90%, and optimum is identical at least 95%.

Here used " compsn " refers to any mixture.Can be solution, suspension, liquid, powder, ointment, water-based, nonaqueous or their any combination.

Here used " associating " refers to any associating between two or more.

Term used herein " object " comprises humans and animals, for example, and dog, cat, ox, pig, rodent etc., preferred people.It is to be suitable for and to be ready the middle-aging male male sex hormone that is caused by male sex hormone and/or androgen receptor functional disorder or follow partly lacked diseases such as syndrome, osteoporosis, muscle wasting, apocleisis, female hirsutism, the alopecia of severe male sex hormone dependent form, acne, evil matter disease, benign prostatic hyperplasia, prostate cancer, mammary cancer, alzheimer's disease and parkinsonism and illness and to treat and prevent that experienced implementer should understand object.

Any protectiveness group used herein, the abbreviation of amino acid and other compounds, consistent with their abbreviations general, that generally acknowledge or the biochemical name of IUPAC-IUB council promulgation, unless stated otherwise.

Prescription and dosage

According to the present invention; Compound of the present invention is united separately or with other medicament, carrier or vehicle; For any suitable route of administration is formulated preparation, for example intracavitary administration, subcutaneous injection, intravenous injection, intramuscularly, intradermal are injected, oral or local application.Present method can be used injecting and administering preparations, with the form of single dose at ampoule, or in the multi-dose container with the buffer reagent drug administration by injection that adds.Preparation can be taked following form such as suspension, solution or the emulsion in oiliness or aqueous media.Preparation can contain prescription reagent such as suspensoid, stablizer and/or dispersion agent.In addition, before the use, activeconstituents can powder type and suitable carriers, aseptic no heat source water or other solvents formation formulation.Local application of the present invention can adopt foam, gel, and ointment, ointment changes leather diaphragm, or paste.

Operable medicinal compsns and the method that is used for administration includes, but are not limited among the present invention, USP 5,736,154,6,197; 801B1,5,741,511,5,886,039,5,941; 868,6,258,374B1 and 5,686,102 contents reported.

The dosage size of treatment or prevention due to illness the seriousness of feelings with route of administration and variation to some extent.Dosage can react different because of age, body weight, healthy state and individual patient with the medication frequency.

It is to be noted (the diagnosis and treatment doctor also should know), according to toxicity and side reaction, the termination of must taking the necessary measures, interruption or reduction therapeutic dose.On the contrary, if clinical response not obvious (getting rid of toxicity and side reaction), the doctor should suitably adjust regimen, improves dosage.

Any suitable route of administration all may be utilized.Formulation comprises tablet, lozenge, beans shape capsule, dispersion agent, suspension agent, solution, capsule, diaphragm and analogue etc.

In practical application, compound of the present invention is united separately or with other preparations, can be according to general pharmacology hybrid technology and pharmaceutical carrier or vehicle, and for example beta-cyclodextrin and 2-hydroxyl-propyl group-beta-cyclodextrin are closely mixed.According to the needs of dispensing, can adopt the special carrier of universal support, part or parenteral route.Prepare non-parenteral dosage forms, for example intravenous injection or the compsn inculcated can adopt similar medicine medium, water known in those skilled in the art, terepthaloyl moietie, oil, buffer reagent, sugar, sanitas, liposome etc.The example of this non-enteron aisle compsn comprises, but is not restricted to Vadex, saline water or other solution of 5%W/V.The total dose of compound of the present invention, separately or with other preparation administation of combination, the administration of available bottle intravenous fluid, volume is approximately from 1 milliliter to 2000 milliliters.According to the total dose of administration, the dilution liquid measure also can be different.

The present invention also provides the medicine box of realizing regimen.This medicine box is united the compound of the present invention of effective dose separately or with other reagent with acceptable form on the pharmacology, is included in one or more containers.Preferably medicament forms is and Sterile Saline, dextrose solution, and buffered soln, or other drug is learned upward, and acceptable sterile liquid share.Perhaps, compsn can be by freeze-drying or drying; In this case, medicine box is randomly further with acceptable solution on a kind of pharmacology, and preferred aseptic solution is included in the container, is formed for injecting the solution of purpose to reformulate mixture.Acceptable solution is salt brine solution and dextrose solution on the typical pharmacology.

In another embodiment, medicine box of the present invention further comprises and is used for the preferred with the pin of sterile form packing or the alcohol pads of syringe and/or packing of injectable composition.Can randomly comprise the specification sheets that supplies doctor or patient to use.

Below in conjunction with specific embodiment the present invention is done further elaboration, but do not limit the present invention.

Embodiment 1

Synthesizing of 3-phenyl-3-(4-oil of mirbane amido)-1-(4-chloro-phenyl-)-1-acetone (MWW6003)

Phenyl aldehyde 10.60 grams (0.1mol), 4-N-methyl-p-nitroaniline 13.81 grams (0.1mol), absolute ethyl alcohol 150ml are added in the reaction flask; After the stirring at room 10 minutes; Add the concentrated hydrochloric acid of 4-chloro-acetophenone 15.46 grams (0.1mol) and catalytic amount, the stirring at room reaction is 20 hours then.After reaction finishes, with the reaction solution cool overnight, the solid that suction filtration is separated out, and use absolute ethanol washing.The gained solid suspension is in 150ml95% ethanol, and stirring at room 2 hours is used saturated NaHCO ₃Neutralization solution continues to stir 1 hour to alkalescence, suction filtration, and with a small amount of absolute ethanol washing filter cake, bullion gets needle crystal 29.09 grams, yield 76.4% through ethanol/water mixed solvent (1:1) recrystallization.

¹HNMR(CDCl ₃)：8.00(2H，d，J＝9.0Hz，Ar-H)，7.81(2H，d，J＝8.1Hz，Ar-H)，7.42(2H，d，J＝8.1Hz，Ar-H)，7.23-7.36(5H，m，Ar-H)，6.52(2H，d，J＝8.7Hz，Ar-H)，5.08(1H，t，J＝6.3Hz，CH)，3.49(2H，d，J＝6.0Hz，CH ₂)；MS(FAB)：382(M+H)。

Embodiment 2

Synthesizing of 3-phenyl-3-(4-oil of mirbane amido)-1-(4-bromophenyl)-1-acetone (MWW6015)

Phenyl aldehyde 10.60 grams (0.1mol), 4-N-methyl-p-nitroaniline 13.81 grams (0.1mol), absolute ethyl alcohol 150ml are added in the reaction flask; After the stirring at room 10 minutes; Add the concentrated hydrochloric acid of 4-bromoacetophenone 19.91 grams (0.1mol) and catalytic amount, the stirring at room reaction is 20 hours then.After reaction finishes, with the reaction solution cool overnight, the solid that suction filtration is separated out, and use absolute ethanol washing.The gained solid suspension is in 150ml95% ethanol, and stirring at room 2 hours is used saturated NaHCO ₃Neutralization solution continues to stir 1 hour to alkalescence, suction filtration, and with a small amount of absolute ethanol washing filter cake, bullion gets needle crystal 30.66 grams, yield 72.1% through ethanol/water mixed solvent (1:1) recrystallization.

¹HNMR(CDCl ₃)：8.00(2H，d，J＝9.3Hz，Ar-H)，7.73(2H，d，J＝8.4Hz，Ar-H)，7.58(2H，d，J＝8.4Hz，Ar-H)，7.23-7.39(5H，m，Ar-H)，6.52(2H，d，J＝9.3Hz，Ar-H)，5.09(1H，t，J＝6.0Hz，CH)，3.49(2H，d，J＝7.2Hz，CH ₂)；MS(FAB)：426(M+H)。

Embodiment 3

Biological activity test

1. material installation

1.1 plasmid and cell strain: expression of androgen receptor plasmid and luciferase reporter gene plasmid are made up by The National Center for Drug Screening; Human cervical carcinoma's epithelial cell strain HeLa, human breast cancer cell strain MDA-MB-453 and Human Prostate Cancer Cells strain LNCaP are all available from U.S. ATCC.

1.2 reagent: foetal calf serum (Fetal bovine serum, FBS, GIBCO/BRL, USA); Gac and VISOSE processing foetal calf serum (CD-FBS, Hyclone, USA); DMEM and RPMI1640 substratum (GIBCO/BRL, USA); The IMEM substratum (Bioresource, USA), the luciferase detection kit (Promega Corporation, USA); Fugene6 (Roche Ltd., USA); [ ³H] and androstanediol (Dehydrotestosterone, DHT, Amersham, UK); Scintillation solution (SuperMixTM, PerkinElmer, USA); Androgen receptor protein is the expression product of this receptor gene in insect cell.

1.3 instrument: Envision2101Multilabel Reader (PerkinElmer, USA); CO2gas incubator (Forma, USA); Wallac

TriLux1450 (PerkinElmer, USA); VERSA ^MaxMicroplate Reader (Molecular Devices, USA).

2. experimental technique and result

2.1 receptors bind vigor test

With the DHT shown in the DMSO preparation table 1 and the gradient solution of each compound of the present invention; DHT concentration is followed successively by 0,0.3,1,3,10,30,100nM; Compound concentration is followed successively by 0,0.128,0.64,3.2,16,80,400,2000nM, adds respectively in each gradient DHT of 5 μ l or each hole of testing compound solution to comb.Androgen receptor protein is added damping fluid [the 25mM NaPO4 that is furnished with 1 μ g/ μ l Trypsin inhibitor,Trasylol (Aprotinin) and leupeptin proteinase inhibitor such as (Leupeptin) in advance; 10% glycerine (Glycerol), 10mM NaMoO4,10mM KF; PH7.5] in, add final concentration and be 5nM [ ³H] DHT, fully behind the mixing, the amount by every hole 195 μ l adds in the comb 4 ℃ of incubated overnight rapidly.Hatch finish after, in the every hole of comb, add 50 μ l Win 40350 (HA) solution (25%, 25mM Na3PO4, pH=7.4), the concussion mixing was hatched 10 minutes, concussion in per therebetween three minutes once.Centrifugal 3 minutes of 2500rpm siphons away supernatant, keeps deposition.Every hole adds the above-mentioned damping fluid of 200 μ l, avoids the vibration deposition, recentrifuge 3 minutes as far as possible.Siphon away supernatant; Keep deposition, repeated centrifugation 1 time siphons away supernatant; Keep deposition; Every hole adds 300 μ l scintillation solutions, and the concussion mixing is with Wallac

TriLux1450 reading.Two compounds are suitable with positive drug DHT to the avidity of androgen receptor, its IC ₅₀Value is less than 10nM (seeing Fig. 1, table 1).

Table 1 compound is to the combination vigor test of androgen receptor

2.2 cytotoxicity experiment

The HeLa cell cultures is waited to grow to 90% and is merged, with adding 96 orifice plates with 4000/ hole after the trysinization, 37 ℃ of overnight cultures in the RPMI1640 substratum that contains 10%FBS and 2mM L type glutaminate (L-glutamine).Testing compound is added cell with finite concentration dilution back, and the concentration of compound MWW6003 and MWW6015 is followed successively by 0,2.56, and 12.8; 64.0,320,1600; 8000,40000nM adds MTT solution (5mg/mL) behind the cultivation 68h; The 560nm absorbance value is surveyed in 20 μ L/ holes, and reference wavelength is 690nM.Experimental result shows that compound MWW6003 and MWW6015 only have low cytotoxicity, the IC of MWW6003 to the HeLa cell ₅₀Value is 2.8 μ M, the IC of MWW6015 ₅₀Value is (see figure 2) between 10～40 μ M.

2.3 reporter gene expression detects

The MDA-MB-453 cell cultures is in the IMEM substratum that contains 10%FBS and 2mM L type glutaminate (L-glutamine).Transfection changes the IMEM substratum that contains 5%CD-FBS into previous day, and Fugene6 reagent is adopted in transfection.Reporter gene carrier and Fugene6 are evenly dropwise added in the cell with the mixed of 1:3, at 37 ℃ and 5%CO ₂Cultivated 6 hours under the condition.Insert 96 well culture plates with 20000/100 μ l/ holes behind the cell dissociation, cultivate based on 37 ℃ with the IMEM that contains 5%CD-FBS and cultivated 2 hours.Add testing compound.During the agonist mode detection, DHT, MWW6003 and MWW6015 concentration are followed successively by 0,0.01,0.1,1,10,100,1000,10000nM; (Bicalutamide BCA) is followed successively by 0,0.01,0.1,1,10,100,1000 with compound concentrations to bicalutamide during the antagonism mode detection, and 10000nM adds 2nM DHT simultaneously as agonist in the cell.After hatching 24 hours, use the luciferase detection kit and detect enzymic activity, assess the pharmacologically active of compound androgen receptor with this.Compound MWW6003 and MWW6015 be display part AR agonist activity under activation pattern, and its usefulness is respectively 19.5% and 20.4% of DHT, EC ₅₀Be respectively 320.1nM and 159.6nM, the EC of DHT under the similarity condition ₅₀Be 0.6nM; Compound MWW6003 and MWW6015 have all shown certain AR antagonistic activity, its IC under the antagonism pattern ₅₀Be respectively 884.8 and 958.1nM, and the IC of BCA under the similarity condition ₅₀Be the 690.9nM (see figure 3).

2.4 prostate cancer cell strain propagation detects

The LNCaP cell cultures is in the RPMI1640 substratum that contains 10%FBS and 2mM L type glutaminate (L-glutamine).Experiment changes the RPMI1640 substratum that contains 5%CD-FBS into previous day, waits to grow to 90% and merges, with adding 96 orifice plates with 4000/90 μ L/ hole after the trysinization, 37 ℃ of overnight cultures.Testing compound is added cell with finite concentration dilution back with 10 μ L/ holes, and during the agonist mode detection, DHT, MWW6003 and MWW6015 concentration are followed successively by 0,0.01,0.1,1,10,100,1000,10000nM; BCA and compound concentrations are followed successively by 0,0.01,0.1,1,10,100,1000 during the antagonism mode detection, and 10000nM adds 5nM DHT simultaneously as agonist in the cell.Cultivated 6 days, and changed dressings once in the time of the 3rd day for 37 ℃.Cultivate and finish the preceding MTT solution (5mg/mL) that adds, the 560nm absorbance value is surveyed, reference wavelength 690nm in 20 μ L/ holes.Experimental data is as shown in Figure 4, and compound MWW6003 and MWW6015 all show certain activity trend (Fig. 4) under the pattern of exciting and antagonism.

3. experiment conclusion

(1) compound MWW6003 and MWW6015 show that combination is active preferably, its IC in the competitive combination vigor test of androgen receptor ₅₀Numerical value is less than 10nM, and is suitable with DHT.

(2) compound MWW6003 and MWW6015 demonstrate low cytotoxicity in the toxicity test of HeLa cell.

(3) compound MWW6003 and MWW6015 show to have androgen receptor excitement/antagonistic activity preferably in MDA-MB-453 cell reporter gene detection method; In the LNCaP proliferation experiment, also shown certain excitement/antagonistic activity trend, pointed out it, the purposes in preventing/treating male sex hormone and androgen-receptor related diseases as androgen modulators.

Claims (3)

1. one type of compound with following general formula I structure or its pharmacy acceptable salt prevent and/or treat symptom that male sex hormone and androgen receptor functional disorder cause or the purposes in the disease medicament as the nonsteroidal androgen receptor agonist in preparation:

Wherein, R is Cl or Br.

2. purposes according to claim 1; It is characterized in that described pharmacy acceptable salt is compound and hydrochloric acid, Hydrogen bromide, nitric acid, sulfuric acid, phosphoric acid, formic acid, acetate, propionic acid, phenylformic acid, toxilic acid, fumaric acid, succsinic acid, tartrate, Hydrocerol A, the alkylsulphonic acid of general formula I structure, the salt of aryl sulfonic acid.

3. claim 1 or 2 described purposes, symptom that wherein said male sex hormone and androgen receptor functional disorder cause or disease comprise that the middle-aging male male sex hormone lacks partly that syndrome, osteoporosis, muscle wasting, femal sexual function are complete, apocleisis, evil matter are sick, alzheimer's disease and parkinsonism.

Images