Background technology
Osteoporosis is worldwide, more and more a to attract people's attention health problem.The current whole world about 200,000,000 people suffers from osteoporosis, its sickness rate leapt to common disease, the 7th of frequently-occurring disease.Estimate that ill total number of persons is more than 200,000,000.Osteoporosis is the modal disease of the elderly.In the U.S. and western countries, more than 55 years old there is the people of half to suffer from the different osteoporosis of degree, and suffer from osteoporosis person up to more than 95% in the elderly of over-65s in postclimacteric women at the age.Along with the progress of the reach of science and the mankind, the average life span constantly extends, and elderly population increase sharply, and osteoporotic sickness rate is also along with appearance increases rapidly.Expectation will be increased to 15.55 hundred million by current 3.23 hundred million to the elderly of the year two thousand fifty whole world over-65s, osteoporosis causes hip fracture generation number to be also increased to 6,260,000 by by current 1,660,000 when the time comes, and wherein the patient of Asia, Latin America, the Middle East and African country will account for more than 70%.According to official statistics, the U.S. reaches 10,000,000,000 dollars for the direct and indirect cost of osteoporosis aspect every year; The fund that the health of Britain and social security institution provide for osteoporosis is every year more than 500,000,000 pounds; France only pays hospitalization cost about 13.5 hundred million French Franc of the patient of 30,000 Hip fracture every year on average.The so huge society that osteoporosis causes and economical load, constitute a serious global problem, caused the very big concern of countries in the world.The arrival of aging society makes osteoporotic incidence increasingly increase, the quality of life of serious threat people.
China is the country that world population is maximum, and wherein elderly population account for 1/2 of Asia elderly population and 1/5 of world population.The osteoporosis epidemiology survey result display in Beijing, Shanghai and Chengdu 3 city, 60-69 year the osteoporotic incidence women of age bracket and the male sex be respectively 60% and about 30%.China's large population base, sufferers of osteoporosis face has reached 6,000 ten thousand ~ 8,000 ten thousand examples, and this brings serious burden to family and society.People according to another statistics national over-65s in 2000 has 1.3 hundred million, accounts for 10.7% of national population, has become typical veteran form country.And the patient suffering from osteoporosis accounts for 90% in these crowds, and the patient of hip fracture will have 12% ~ 40% to die from various complication in 1 year.Also the human action inconvenience of 50% is had in survivor.This not only causes serious body and mind to patient and wrecks, and adds huge manpower and financial resources burden also to family and society simultaneously.Therefore, it is anti-ageing for preventing and treating osteoporosis, prolongs life, ensures the research topic that of quality of life of the people is very urgent.
Summary of the invention
The object of this invention is to provide one and treat osteoporotic pharmaceutical composition.
In order to realize object of the present invention, the invention provides one and treat osteoporotic compound, this compound has having structure:
The present invention also provides one to treat osteoporotic pharmaceutical composition, and described pharmaceutical composition includes the compound of effective amount and pharmaceutically acceptable carrier, and described compound has having structure:
Preferably, described pharmaceutically acceptable carrier is thinner, disintegrating agent, tackiness agent, lubricant, stablizer or corrigent.
Preferably, described thinner is sugar derivatives, starch derivative or derivatived cellulose.
Preferably, described thinner is lactose.
Preferably, described pharmaceutical composition is powder, microgranules, granule, capsule or tablet.
The present invention also provides the purposes of compound in the osteoporotic medicine of preparation treatment, and this compound has having structure:
Term used herein " pharmaceutically acceptable " refers to not eliminate the biologic activity of compound as herein described or the material of character, as carrier or thinner.This kind of material is applied to and individual does not cause undesirable biological action or not with harmful way and any component interaction comprised in its composition.
" pharmaceutically acceptable carrier " comprises any and all solvents as the term is employed herein, dispersion medium, coating material, tensio-active agent, antioxidant, sanitas (such as antiseptic-germicide, anti-mycotic agent), isotonic agent, absorption delay agent, salt, sanitas, drug stabilizing agent, tackiness agent, vehicle, disintegrating agent, lubricant, sweeting agent, correctives, dyestuff etc. and its combination, this is well-known to those skilled in the art (for example, see Remington'sPharmaceutical Sciences, 18th Ed.Mack PrintingCompany, 1990, pp.1289-1329).Except with except the inconsistent carrier of activeconstituents, consider to use any conventional carrier in treatment or pharmaceutical composition.
Compound of the present invention all shows certain promoter action to the propagation of rat kind skeletonization UMR 106 cell and/or differentiation.
Embodiment
The present invention is further illustrated below by embodiment.It should be understood that embodiments of the invention are for illustration of the present invention instead of limitation of the present invention.Essence according to the present invention all belongs to the scope of protection of present invention to the simple modifications that the present invention carries out.Unless otherwise stated, the percentage ratio in the present invention is weight percentage.
Experimental example
Material: α-MEM substratum is Gibco Products, and Hepes trypsinase is Sigma Products, UMR106 cell strain, purchased from Peking University's oral surgery laboratory, comes from masschusetts, u.s.a hospital general.Foetal calf serum is purchased from hemopathy institute of the Chinese Academy of Medical Sciences.
Method: with rat human osteosarcoma cell system UMR106 for target cell, adopt the method for controlled observation, take estradiol as positive control, establish Normal group (not dosing simultaneously, only add equal cell suspension and substratum) and blank group (not dosing and cell suspension only add equivalent substratum).Preparation compound is 1 × 10
-8, 1 × 10
-7, 1 × 10
-6mol/L 3 concentration groups, totally 6 groups.Preparation positive control and compound use DMSO.
UMR106 cell cultures and morphological observation: by UMR106 cell cultures in the α-MEM substratum containing 150mL/L foetal calf serum, 25mmol/L Hepes, 100kU/L penicillin, 100mg/L Streptomycin sulphate, put 37 DEG C, cultivate in 50mL/L CO2gas incubator.Get the cell that growth conditions is good, use 2.5g/L tryptic digestion, add α-MEM substratum and make cell suspension, adjustment cell concn is 4 × 10
7l
-1, be inoculated on 4 piece of 12 well culture plate, in culture plate, place 1.5 × 1.5cm slide.Blank group, estradiol positive controls, 1 × 10
-8, 1 × 10
-7, 1 × 10
-6mol/L 3 concentration groups respectively establish 8 multiple holes, and every hole adds cell suspension 2.7mL, corresponding liquid 0.3mL.After 48h, observation of cell metamorphosis under inverted microscope.
The compounds of this invention is on the impact of UMR106 cell proliferation: adjustment cell concn is 2 × 10
7l
-1, be inoculated in 96 well culture plates, every hole adds cell suspension 180 μ L, after 24h, adds the compound of above-mentioned 3 concentration, every hole 20 μ L, separately establishes blank group (do not add compound, only add equal cell suspension).Often group establishes 8 multiple holes, and after continuing to cultivate 48h, adopt tetrazolium salts method to be measure wavelength with 490nm, 655nm is reference wavelength, measures the absorbance A value in each hole.Calculate average proliferation rate.
The compounds of this invention is on the impact of the inside and outside alkaline phosphatase activities of UMR106 cell: adjustment cell concn is 2 × 10
7l
-1, be inoculated on 2 piece of 24 well culture plate, if Normal group, estradiol 1 × 10
-8l
-1positive controls and compound 1 × 10
-8, 1 × 10
-7, 1 × 10
-63 concentration groups of mol/L, totally 5 groups, often group establishes 8 multiple holes, according to the method for Joha AP, after dosing 72h, adopts p-nitrophenyl matter KINETIC METHOD to measure in cell and the alkaline phosphatase activities of substratum.
MAIN OUTCOME MEASURES: the compounds of this invention 1 × 10
-8, 1 × 10
-7, 1 × 10
-63 concentration groups of mol/L are on the impact of UMR106 cell proliferation and inside and outside alkaline phosphatase activities.
Statistical analysis: completed by SPSS statistical software.Data are used
represent, t inspection between organizing, namely P<0.05 thinks to have significant difference.
Result
The impact that the compound of different concns changes UMR106 cellular form
After adding compound cultivation 48h in substratum, cell quantity showed increased compared with control group, karyokinetic phase, is common.Normal group is after cultivation 48h, and control group UMR106 cell proliferation is slow, has no matrix and piles up.Add compound 1 × 10
-8after mol/L cultivates 48h, obviously, cell count increases UMR106 propagation, and a small amount of matrix is piled up.Add compound 1 × 10
-7after mol/L cultivates 48h, cell count obviously increases, and overlapping growth, matrix is piled up obviously.Add compound 1 × 10
-6after mol/L cultivates 48h, cell is bred in a large number, connects in flakes, matrix bulk deposition, overlapping growth.
The proliferation of the compounds of this invention to UMR106 cell sees the following form
Cell is after compound treatment 48h, and detect through tetrazolium salts method, relative to control group, 3 administration groups all have significant difference compared with control group.
Compound is on the impact of alkaline phosphatase activities in UMR106 cell
Cell is after compound treatment when 24h, 48h, and detect through p-nitrophenyl matter KINETIC METHOD, in cellular control unit, alkaline phosphatase activities is respectively 713.12 ± 12.15,2281.32 ± 7.62 μ kat/g, compound 1 × 10
-6in mol/L group cell, alkaline phosphatase activities is respectively 823.12 ± 12.31,2415.41 ± 15.86 μ kat/g, and difference has significant compared with control group.Estradiol 1 × 10
-8mol/L group 24h, 48h after treatment, in cell, alkaline phosphatase activities is respectively 275.19 ± 18.63,2355.48 ± 15.62 μ kat/g, and difference has significant compared with control group.72h after process, each concentration group of compound and estradiol 1 × 10
-8mol/L group group is compared with control group, and in cell, alkaline phosphatase activities has the trend of enhancing, but no difference of science of statistics.
Compound is on the impact of UMR106 extracellular alkaline phosphatase activities
After 24h, control group substratum activity change of Alkaline phosphatase is 652.83 ± 11.31 μ kat/g, compound 1 × 10
-6mol/L group is 781.62 ± 11.58 μ kat/g, has significant difference compared with control group.Estradiol 1 × 10
-8mol/L group and compound 1 × 10
-7mol/L group is respectively 728.85 ± 9.87 μ kat/g and 733.18 ± 15.32 μ kat/g, has significant difference compared with control group.Compound 1 × 10
-8mol/L group is 671.32 ± 14.27 μ kat/g, with control group indifference.48h and 72h after treatment, each concentration group of compound and estradiol 1 × 10
-8mol/L group is compared with control group, and extracellular alkaline phosphatase activities has the trend of enhancing, but no difference of science of statistics.