CN1718223A - Medicine for treating osteoporosis contg. astraglus base and stanozolol - Google Patents

Abstract

A medicine in the form of pill, tablet, particle, capsule, injection, or exterior-applied cream for treating osteoporosis is prepared from the dried root of Astragalus membranaceus (Fisch.)Bge. mongholicus (Beg) Hsiao or Astragalus membranaceus (Fish.)Bge and one of the assimilative hormones.

Description

A kind of pharmaceutical preparation of the treatment osteoporosis of forming by the Radix Astragali and stanozolol

Technical field the present invention relates to be used for the treatment of the pharmaceutical composition of osteoporosis, it is characterized in that this pharmaceutical composition is made up of the anabolic hormone and the Radix Astragali.

Background technology primary osteoporosis (comprising postmenopausal osteoporosis and senile osteoporosis) be a kind of with the bone amount reduce, the osseous tissue fine structure is degenerated, and (the Grafting Cancellous Bone Bolt girder attenuates, rupture, the quantity minimizing; Cortical bone porous, attenuation) be feature, so that the fragility of bone increases the senile skeletal diseases of the general that fracture risk increases.Osteoporosis is a kind of disease, shows as the bone amount and reduces with clinical signs such as increase of skeleton fragility under normal physical activity and skeleton pains.Because the bone amount reduces, the fine structure of bone is degenerated and the fragility of bone increases, cause the biomechanical strength of bone to descend, be difficult to carry the load of original or relative increase, capillary fracture or complete fracture can quietly take place, have a strong impact on old people's quality of life, bring very big burden for patient, family and society.The epidemiological study of osteoporosis shows, women's menopause in 49 years old can be suffered from the bone amount in 52--60 year and be reduced, and 62--76 year can suffer from osteoporosis, and 72--86 year can be suffered from serious osteoporosis.Except that primary osteoporosis, secondary osteoporosis, particularly corticosteroid osteoporosis also faces stern reality, in the patient of the chronic disease of accepting corticosteroid treatment, finally also have 30～50% to develop into osteoporosis, more seriously causing femur head necrosis and Fracture of femur, is the iatrogenic disorder problem that can not be ignored.At any time development and national economy, the raising of living standards of the people, the population life-span prolongs gradually, and following society will become senile society.China more than 60 years old aging population risen to 1.34 hundred million people, according to international standard, China has entered aged society.If add the person in middle and old age's population below 60 years old, its quantity is more considerable.As the osteoporosis and the osteoporotic fracture of one of Senile disease, increasing people is deeply hurt, and become the serious social problem of worldwide concern, China has classified it as one of Senile disease of three big emphasis tackling key problem researchs.WHO works up the global strategy target to osteoporosis, improve the whole world to osteoporotic diagnosis and processing, each university, institute, drugmaker one after another first mate's degree increase funds to quicken the research and development of osteoporosis new drug, therefore preventing and treating senile osteoporosis is to guarantee that people are healthy, a very urgent research topic of improving the quality of living.

Think that at present osteoporosis is a kind of disease of multi-pathogenesis, its basic pathology mechanism is that defective appears in bone resorption and osteoplastic balance in the bone metabolism process, cause the intravital alcium and phosphor metabolization imbalance of people, bone density is reduced gradually and cause the main and age of clinical symptoms, its cause of disease, endocrine regulation, calcium absorption is bad, and limbs are useless to be used and immunity, and factors such as nutrition, heredity are relevant.At in recent years 20 years, osteoporotic Drug therapy strategy and approach mainly were two big classes, and one is the medicine of anti-bone resorption, and two for facilitating the bone medicine.Being anti-bone resorption medicine more than 90% in the clinical use, as controversies in hormone replacement in the elderly, is the academic therapy of treatment postmenopausal osteoporosis.But estrogen causes the danger of genital system tumor and is affirmed [2] that the U.S. has declared that estrogen is one of carcinogen; The diphosphonic acid salt can effectively be treated bone loss due to the bone resorption, but the prolonged application side effect is big, also is difficult for as prophylactic; Calcitonin is remarkable to molten bone osteodynia and osteoporosis effect, but treatment needs individuation, also is subjected to the route of administration restriction, and costs an arm and a leg; The present estrogenic agents of paying close attention to (ERMs) is blocked stimulation to reproductive system owing to kept to the estrogen-like effects of skeleton, has advantage, has 2-3 new drug and enters the clinical III phase and test (U.S.).All these anti-bone resorption agent have all suppressed because of bone resorption increases the bone formation that coupling joins, and the conversion ratio of old bone and new bone is reduced, and prolonged application influences the quality and the mechanical property [3] of bone.At present short bone formation medicine becomes the research focus gradually, short chain parathyroid hormone (PTH) is very significantly short bone formation medicine, it is clinical also to enter the II phase in the U.S., but experimental studies results finds that PTH can increase cortical bone porous (because of increasing the absorption of cortical bone Haversian canal simultaneously), reduces the late result of fracture and does not affirm [4]; Active VitD prolonged application easily causes hypercalcemia and urine calcium disease; Fluoride is the short bone formation medicine that early is used for osteoporosis, but it easily makes osteoid increase and with bone mineralising defective, limited in wide clinical application, and the safety range that mainly comes from it is narrow, and side effect is big, and curative effect needs further textual criticism.

Summary of the invention we by a large amount of animal experiment studies and screening, we find that the compound preparation that anabolic hormone and the Radix Astragali are formed has the effect of excellent prevention osteoporosis.Anabolic hormone mainly is a stanozolol, and other anabolic hormones such as nandrolone phenylpropionate, metandienone, oxymetholone, danazol, capric nandrolone also have effect.Stanozolol is a kind of anabolic hormone of synthetic, because of once tested clinically menopausal women osteoporosis and the primary osteoporosis of being used for a little less than the strong androgenic effect of its promotion albumen anabolic effect, we experimental results show that, this medicine has certain preventive and therapeutic effect to the steroid rats with osteoporosis, it is the synthetic anti-osteoporotic of a kind of promotion bone, owing to promote that in the world the medicine of bone formation and few side effects is also few at present, by contrast, this product is one of less medicine of side effect in this class medicine.But because this product has been used decades clinical as anabolic hormone, a large amount of research datas show the untoward reaction that this product prolonged application still can produce liver, as four kinds of transaminase's risings, jaundice etc., high density lipoprotein (HDL) is reduced, low density lipoprotein, LDL (LDL) increases, thereby increased the danger of coronary heart disease, in addition, stanozolol also can cause water sodium pool to slip, edema etc., these untoward reaction have had a strong impact on stanozolol in the purposes of preventing and treating aspect the osteoporosis.This project researcher is found the compound preparation that Chinese medicine astragalus and stanozolol are formed, can improve the effect of osteoporosis greatly, simultaneously the side effect of stanozolol is obviously reduced, be particularly suitable for numerous gerontal patients and take, elderly men osteoporosis better efficacy.

The approach that solves this task is to realize like this, choose the dry root of leguminous plant Radix Astagali Astragalus membranaceus (Fisch.) Bge.var.mongholicus (Beg) Hsiao or Radix Astragali A.membranaceus (Fisch.) Bge, it is the medicinal part that Chinese medicine is called the Radix Astragali, add 15 times of water by traditional technology, steaming and decocting is three times respectively, each 2 hours, filter, merge three times filtrate, be concentrated into certain concentration, as the water extract of the Radix Astragali.Simultaneously, it is standby to extract (+)-Astragenol extract, astragalus polysaccharides, Radix Astragali total flavones, Radix Astragali saponin by known technology.In addition, select the anabolic hormone stanozolol, nandrolone phenylpropionate, metandienone, oxymetholone, danazol, capric nandrolone and the top Radix Astragali and be extracted into and be grouped into compound recipe, set up the rat osteoporosis model with this research department then and carry out administration research, these are subjected to the pharmacodynamics function of test product osteoporosis comparative observation.

Research approach is as follows:

(1) prescription and the experiment of experiment medicine are grouped as follows:

1, water extraction of astragalus membranaceus+stanozolol

2, (+)-Astragenol extract+stanozolol

3, astragalus polysaccharides+stanozolol

4, Radix Astragali total flavones+stanozolol

5, Radix Astragali saponin+stanozolol

6, water extraction of astragalus membranaceus+nandrolone phenylpropionate

7, (+)-Astragenol extract+nandrolone phenylpropionate

8, water extraction of astragalus membranaceus+metandienone

9, (+)-Astragenol extract+metandienone

10, water extraction of astragalus membranaceus+oxymetholone

11, (+)-Astragenol extract+oxymetholone

12, water extraction of astragalus membranaceus+danazol

13, (+)-Astragenol extract+danazol

14, water extraction of astragalus membranaceus+capric nandrolone

15, (+)-Astragenol extract+capric nandrolone

Result of study shows: above 15 prescriptions all have and prevent the osteoporotic effect of laboratory animal preferably, water extraction of astragalus membranaceus+stanozolol wherein, (+)-Astragenol extract+stanozolol and astragalus polysaccharides+stanozolol are effective, and side effect is less, and water extraction of astragalus membranaceus is pressed experimental rat 1.25g～5g.Kg-1; The compound recipe best results that stanozolol 0.125mg0.125mg0.5mg.Kg-1 forms has good DEVELOPMENT PROSPECT.Other prescriptions also will carry out deep research, adjust the prescription ratio, and effect may be also fine.

More than the compound preparation formed by the Radix Astragali and anabolic hormone, can be made into any in clinical acceptable pill, tablet, granule, capsule, injection, the preparation for external application to skin.These preparations can add drug excipient, as solvent, disintegrating agent, correctives, antiseptic, coloring agent, dispersant, binding agent, thickening agent, lubricant, diluent.

The specific embodiment: embodiment one

1 materials and methods

1.1 40 of 4 monthly age of animal SPF level male SD rats, body weight 235 ± 21g provides (Guangdong Province's laboratory animal certification of fitness numbering: 2002A014) by Guangdong Medical College's Experimental Animal Center

1.2 method

1.2.1 reagent

Prednisolone acetate: raw material, fairy house Pharmacy stock Co., Ltd produces, lot number 981220;

The precious extracting solution GBH of stilbene health bone:

The QKH group: stilbene health high dose, prescription are stanozolol 0.5mg.Kg-1+ astragalus polysaccharides 5g.Kg-1.

The QKM group: dosage in the stilbene health, prescription are stanozolol 0.25mg.Kg-1+ astragalus polysaccharides 2.5g.Kg-1.

The QKL group: stilbene health high dose, prescription are stanozolol 0.125mg.Kg-1+ astragalus polysaccharides 1.25g.Kg-1.

Three kinds of dosage particles are provided by Guangdong Medical College science and technology pharmaceutical developments center.

1.2.2 instrument

LEICA QWIN image analyzer (German Lycra); UV-3010 ultraviolet spectrophotometer (day island proper Tianjin); Inductance coupled plasma direct-reading spectrometer (day island proper Tianjin); AE240 electronic balance (prunus mume (sieb.) sieb.et zucc. Teller-Shanghai branch company of Tuo benefit instrument company).Hydroxyproline determination test kit (Hpy) is purchased and is built up biological reagent company in Nanjing.

1.2.3 medication

40 SD rats are divided into 5 groups at random, and 8 every group, 1 group is normal control group (NS group), give NS and handle; All the other 4 groups are all given prednisone by 2.7mg.kg ^-1Give N.S, GBH, GBM and GBL filling stomach more respectively, 5ml.kg after irritating stomach ^-1, every day 1 time, 12w continuously.Weigh weekly in the experimentation, all animals are finished preceding 13,14 days cervical region subcutaneous injection quadracyclines (25mg/kg) conduct fluorescent labeling for the first time in experiment, and experiment finishes preceding 3,4 days injection calcein (5mg/kg) conduct fluorescent labeling for the second time.

1.2.4 testing index

(1) observation of the static index of rat tibia epimere spongy bone norphometry mathematic(al) parameter

Adopt semi-automatic image digitization analyser the thick not stained of 8 μ m of rat tibia epimere undecalcified bone to be measured and calculated the static parameter of bone according to document [2～3], comprise bone trabecula area percent (%TbAr), bone trabecula quantity (Tb.N), bone trabecula thickness (Tb.Th) and bone trabecula separating degree (Tb.Sp), and in 5 μ m section (through Masson-Goldner Trichrome dyeing) number (Oc.N/BV) and the absorption girth percent (%OC.Pm) of the long-pending osteoclast of the meter unit of calculating surface of bone, with the variation of observation bone amount and bone structure and understand its possible cause.

(2) observation of rat tibia epimere spongy bone morphometry dynamic parameter

Adopt semi-automatic image digitization analyser that the bone dynamic parameter is measured in the thick section of 8 μ m of rat tibia epimere undecalcified bone, comprise bone trabecula fluorescent perimeter percentage rate (%L.Pm), mineralising deposition (MAR), bone formation rate (BFR/TV), with the amount and the speed of understanding bone surface mineralising, and explain the possible cause that static parameter changes.

(3) observation of rat bone biochemical indicator

Be baked to constant weight (BW) after muscle and soft tissue are adhered in left side ulna Ex-all, get filtrate after bone specimen is placed 6mol/L HCl constant temperature digestion 24h.Filtrate is divided into two parts, a content of elements of measuring, another part filtrate is measured the content of hydroxyproline.

(4) observation of rat bone physical index

Left side femur Ex-all is adhered to behind muscle on the bone and the soft tissue weighing weight in wet base (BWW) back in baking box dry to constant weight (BDW).Recording greater trochanter of femur and neck of femur meet minimum point with band table slide gauge is the femur width (BD) that femur length (BL) and third trochanter come downwards to femoral shaft handing-over part to the distance of fossa intercondylaris femoris.The macroscopic view growth of bone comprises the long and long thick of bone, therefore measures the overall growth situation that the two can embody bone.

1.3 statistical procedures

All testing indexs all adopt the form of mean plus-minus standard deviation to represent that (x ± s), data analysis uses SPSS 11.0 to analyze, and with p＜0.05, p＜0.01 expression has statistical significance.

2 results

2.1 stilbene health bone treasured is to the influence of prednisone rat tibia epimere spongy bone morphometry static parameter

Each is organized the variation of rat tibia epimere spongy bone morphometry static parameter and sees Table 1:

Table 1 stilbene health bone treasured to the influence of prednisone rat bone morphometry static parameter (x ± s, n=8)

Group	％Tb.Ar (％)	TbN (mm -1)	TbTh (um)	TbSp (um)
					NS GC GBH GBM GBL	15.07±1.70 8.83±0.98 ** 15.89±4.15 △△ 15.45±4.90 △△ 14.31±2.60 △△	2.92±0.23 1.98±0.20 ** 3.07±0.15 △△ 2.92±0.59 △ 2.95±0.43 △	51.86±6.48 44.47±2.43 * 51.50±8.39 △△ 52.72±6.98 △ 49.10±2.58	292.88±25.52 463.53±54.54 ** 281.58±56.37 △△ 299.78±59.58 △△ 301.6±55.3 △△

Compare with NS ^*：P＜0.05， ^**：P＜0.01；Compare with GC △：P＜0.05，△△：P＜0.01

With NS group relatively, %Tb.Ar, Tb.N, Tb.Th 41.3%, 31.9% and 14.2% (P＜0.01) that descends respectively in the GC group rat bone static parameter, prompting GC group rat spongy bone has obviously to be lost.With GC group relatively, three groups of QKGB have improved 89.6%, 84.3%, 70.7% with %Tb.Ar, Tb.N and the Tb.Th of prednisone rat respectively; 55.1%, 47.5%, 49.0% and 15.8%, 18.5% and 10.7%.With Tb.SP, reduced by 39.4%, 35.3% and 34.9.

2.2 stilbene health bone treasured is to the influence of prednisone rat tibia epimere spongy bone morphometry dynamic parameter

Each bone morphometry dynamic parameter variation of organizing rat sees Table 2:

Table 2 stilbene health bone treasured to the influence of prednisone rat spongy bone morphometry dynamic parameter (x ± s, n=8)

Group	％L.Pm (％)	MAR (um/d)	BFR/TV (％/year)	Oc.N/BV (mm -2)	％Oc.Pm (％)
						NS GC GBH GBM GBL	31.88±8.19 29.81±9.02 30.42±7.81 29.67±9.81 29.04±9.01	0.85±0.16 0.81±0.25 0.94±0.21 0.85±0.26 0.78±0.30	49.29±17.74 29.67±13.17 * 46.32±9.62 △ 37.84±8.93 △ 39.51±9.44 △	11.42±1.46 38.38±8.65 ** 10.97±3.36 △△ 12.82±3.33 △△ 13.69±4.65 △△	0.873±0.09 2.54±0.34 ** 0.93±0.03 △△ 1.17±0.02 △△ 0.95±0.03 △△

Compare with NS ^*：P＜0.05， ^**：P＜0.01；Compare with GC △：P＜0.05，△△：P＜0.01.

Compare with the NS group, the variation little (P＞0.05) of %L.Pm and MAR in the GC group rat bone dynamic parameter, but BFR/TV reduces by 40% (P＜0.05), and prompting excusing from death reason for a long time dosage uses prednisone can make the rat Grafting Cancellous Bone Bolt form the total amount minimizing; Simultaneously, Oc.N/BV, the %Oc.Pm of GC group increased by 236.1% and 191.4% (P＜0.01) respectively than the NS group, points out this group osteoclast to increase, and bone resorption increases, and bone loss is obvious.Compare with the GC group, the bone formation rate of three groups of rat spongy bone of QKGB obviously rises, GBH, GBM and GBL organize 56.1%, 27.5% and 33.2% (P＜0.05) of having risen than GC respectively for three groups, and Oc.N, %Oc.Pm have reduced by 63.4%, 53.9% and 62.6%, and prompting QKGB can significantly promote rat bone to form and the absorption of inhibition bone.

2.3 stilbene health bone treasured is to the influence of prednisone rat ulna biochemical indicator

Each variation of organizing the rat bone biochemical indicator sees Table 3:

Table 3 stilbene health bone treasured to the influence of prednisone rat bone biochemical indicator (x ± s, n=8)

Group	UW (g)	Hyp (mg/g)	Ca 2+ (mg/g)	P 3+ (mg/g)
					NS GC GBH GBM GBL	0.168±0.007 0.155±0.010 * 0.165±0.014 △ 0.164±0.010 △ 0.094±0.034 △	40.59±3.6 34.61±6.4 * 36.10±2.5 * 34.01±3.4 * 36.96±5.3 *	212.21±19.7 203.62±24.5 209.48±29.7 190.28±6.97 194.45±17.59	90.23±6.91 99.46±12.9 94.61±10.2 89.45±10.3 92.27±11.3

Compare with NS ^*：P＜0.05， ^**：P＜0.01；Compare with GC △：P＜0.05，△△：P＜0.01.

With NS group relatively, GC group rat ulna weight obviously descends, the Hyp content of unit bone weight also descends but bone calcium, phosphorus content there was no significant difference (P＞0.05).Can cause the organic matter and the bone mineral of bone to lose simultaneously but losing obviously behind the prompting rat prolonged application prednisone with organic matter; Compare with the GC group, QKGB can improve the concentration of bone Hyp, but unit sclerotin content not high (P＞0.05), this may raise relevant with three groups bone representation work.

2.4 stilbene health bone treasured is to the observation of prednisone rat femur physical index

Each bone physical index variation of organizing rat sees Table 4:

Table 4 stilbene health bone treasured to the influence of prednisone rat femur physical index (x ± s, n=8)

Group	BWW (g)	BL (mm)	BD (mm)
				NS GC GBH GBM GBL	0.872±0.04 0.785±0.07 ** 0.862±0.07 △ 0.886±0.11 △△ 0.943±0.11 △△	33.44±0.43 32.64±0.51 * 33.79±0.86 △ 33.38±0.91 △ 33.24±0.68 △	4.43±0.16 4.33±0.23 * 4.50±0.17 △ 4.56±0.39 △ 4.40±0.87 △

Compare with NS ^*：P＜0.05， ^**：P＜0.01；Compare withGC △：P＜0.05，△△：P＜0.01.

Compare with the NS group, GC group rat femur weight in wet base (BWW), femur dry weight (BDW), femur length (BL) and femur width (BD) obviously reduce (P＜0.05), behind the prompting prednisone life-time service rat sclerotin is obviously lost, can significantly suppress simultaneously the growth of bone, show as the long and long slightly limited of bone, especially limited obviously with length length.After the QKGB treatment, the bone of three groups of rats is heavy, length and width all increase (P＜0.01) than the GC group, and prompting CD can obviously resist the osteogenesis inhibitory action of prednisone.

3 discuss

Originally experimental studies have found that, prednisone is had children outside the state plan the reason dose application for a long time can make the rat bone amount obviously lose, the sclerotin balance is destroyed, show as mainly that Hyp content significantly reduces in the bone, and calcium and phosphorus content variation not obvious (P＞0.05) in the unit bone, but calcium in the unit bone/phosphorus ratio reduction, content organic in the bone is lost degree greater than inanimate matter, cause the sclerotin component ratio inharmonious, inanimate matter content is relatively than content of organic matter height.Hydroxyproline (Hyp) is the special composition in the ossein, content is more stable in ossein, it is that the minimizing of organic its content of sign that reduces in the bone can cause the fragility of bone to increase and toughness descends that its content reduces, and this may be the prolonged application glucocorticoid patient reason of fracturing easily clinically.GC group rat BFA/TV reduces in this experiment, and the bone formation of prompting unit osseous tissue area reduces, and sclerotin content significantly reduces.%Tb.Ar is the important indicator of reflection bone quantitative changeization, and Tb.N, Tb.Th and Tb.SP are the microstructural aggregative indicatores of reflection bone trabecula, and experimental result shows that prednisone uses the above parameter generation of back rat significant change, presents the osteoporosis state.

The stilbene recovering capsule is made up of astragalus polysaccharides and stanozolol (stanozolol), and in prescription, the Radix Astragali can not suppress the bone loss that bone formation is used for resisting castrated rats by suppressing bone resorption ^[4]Research is thought ^[5～6]Radix Astragali defying age, enhancing immunity and to improve the effect of blood circulation protection hepatic and renal function outstanding also can promote osteoblast secretion osteoid by promoting collagen protein synthesis ^[7], the function of osteoblast inhibitory action that can effectively cause to the Antiglucocorticoid prolonged application.Stanozolol is a kind of assimilation parahormone of synthetic, can promote proteinic synthesizing, and can increase calcium deposition, promotes the synthetic effect of bone, the Wu Tie of this seminar etc. ^[8]Once proved that it lost the prednisone rat bone tangible preventive and therapeutic effect is arranged, treatment castration and drug-induced osteoporosis disease were had certain curative effect, also observed the osteoporosis symptom that stanozolol can obviously improve the patient clinically ^[9]This experimental result also points out the stilbene recovering capsule of being made up of these two kinds of Chinese medicines to have the osteoporotic effect of better prevention glucocorticoid.

Claims (4)

1, the pharmaceutical composition that is used for the treatment of osteoporosis is characterized in that this pharmaceutical composition is made up of the anabolic hormone and the Radix Astragali, and anabolic hormone can be stanozolol, nandrolone phenylpropionate, metandienone, oxymetholone, danazol, capric nandrolone.

2, the pharmaceutical composition that is used for the treatment of osteoporosis as claimed in claim 1, the Radix Astragali of said composition can be water extract, the alcohol extract of the Radix Astragali, astragalus polysaccharides, Radix Astragali total flavones, the Radix Astragali saponins of the Radix Astragali.

3, arbitrary pharmaceutical composition as claimed in claim 1 is characterized in that its obtained preparation is any in clinical acceptable pill, tablet, granule, capsule, injection, the preparation for external application to skin.

4, arbitrary pharmaceutical composition according to claim 1 is characterized in that also can adding drug excipient in the middle of its preparation.