CN103251630B - Medical and the medicine for health care of brassinolide - Google Patents
Medical and the medicine for health care of brassinolide Download PDFInfo
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- CN103251630B CN103251630B CN201310086548.8A CN201310086548A CN103251630B CN 103251630 B CN103251630 B CN 103251630B CN 201310086548 A CN201310086548 A CN 201310086548A CN 103251630 B CN103251630 B CN 103251630B
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- 230000036541 health Effects 0.000 title claims abstract description 25
- 239000003814 drug Substances 0.000 title claims abstract description 24
- IXVMHGVQKLDRKH-KNBKMWSGSA-N brassinolide Chemical compound C1OC(=O)[C@H]2C[C@H](O)[C@H](O)C[C@]2(C)[C@H]2CC[C@]3(C)[C@@H]([C@H](C)[C@@H](O)[C@H](O)[C@@H](C)C(C)C)CC[C@H]3[C@@H]21 IXVMHGVQKLDRKH-KNBKMWSGSA-N 0.000 title description 4
- IXVMHGVQKLDRKH-VRESXRICSA-N Brassinolide Natural products O=C1OC[C@@H]2[C@@H]3[C@@](C)([C@H]([C@@H]([C@@H](O)[C@H](O)[C@H](C(C)C)C)C)CC3)CC[C@@H]2[C@]2(C)[C@@H]1C[C@H](O)[C@H](O)C2 IXVMHGVQKLDRKH-VRESXRICSA-N 0.000 title description 3
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- 230000002265 prevention Effects 0.000 claims abstract description 7
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Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides brassinolide analog for the preparation of prevention or treatment mammal (especially boar) disease medicine in application, or for the preparation of health care mammiferous health product in application.In addition, present invention also offers corresponding medicine or Halth-care composition and preparation method thereof.
Description
Technical field
The present invention is the continuation application of No. 201210026285.7th, Chinese patent, belong to for mammal (as, people) medical art, specifically, the present invention relates to the medical of brassinolide and pharmaceutical composition for health care, for the preparation for the treatment of or preventing in the medicine of mammalian diseases.
Background technology
Brassin lactones is called as the sixth-largest class phytohormone in the world, is the growth regulator for plant.Wherein, the brassin lactones of chemosynthesis, as described in Chinese patent application CN1217337A, CN1217338A etc., although product is comparatively homogeneous, kind is few, and mainly 28-shows high brassin lactones.
From plant (as, pollen) or other natural origins (as, Cera Flava) the middle brassin lactones extracted, it normally contains the mixture of multiple brassin lactones analog, the situation of easy generation unstable product quality, even if for the growth regulator for plant like this concerning the comparatively tolerant project of stability, the effect that also more difficult acquisition is stable, more cannot be used for the higher project of requirement (as body of mammals); If purified further, then preparation cost rises comparatively large, is unfavorable for industrialized implementation, thus hinders the further large-scale purification of people, and hinder people and go to study wherein each natural brassinolide analogue whether there is the activity of brassin lactones, more cannot imagine mammiferous medicinal application.
The present inventor passes through painstaking efforts for a long time, find out the stable flow process extracting (purification) natural brassinolide analogue from plant, study intensively wherein each kind of Extraction solvent, although wherein need certain purification process, but only need a leaching process, finally can collect the highly purified brassinolide analog of multiple separation, the cost of purifying of thus having made thinner, is conducive to Industry Promotion and implements; These brassinolide analog be separated can be used alone, also can proportioning is mixed uses, thus constant product quality and easily controlling.
More unexpectedly, on the basis obtaining a large amount of highly purified brassinolide analog be separated, the present inventor chances on, some in these natural natural brassinolide analogue, can be used in as mammal medicine or health product, such as prevention or treatment boar prostatoplasia diseases, or for improving boar prostatic hyperplasia situation.
Summary of the invention
The technical problem to be solved in the present invention there are provided new medicine or the application of health product, can be used for the treatment of mammal (e.g., people), prevention or improvement.In addition, present invention also offers the stable pharmaceutical composition of proportioning or health product, and prepare their method.
Specifically, in first aspect, the invention provides the application of brassinolide analog in the medicine for the preparation of prevention or treatment mammalian diseases; In addition, present invention provides brassinolide analog for the preparation of the application improved in the health product of mammalian condition.
In this article, medicine or health product, refer to the medicine or health product used for mammal (especially people), instead of for the compositions that plant is used.Obviously, the requirement of the medicine used for mammal (especially people) or health product is more much higher than the requirement of the compositions used for plant, and the former needs proportioning to stablize, and the latter can be more unstable; The former needs aseptic, and the latter can some bacterium; The former impurity content is low, and the latter can contain more impurity, so even if the key component of the former with the latter is identical, but both are different, or the former is the part can not selecting by the latter's requirement in latter category to choose.
In addition, the difference of medicine and health product be those skilled in the art be familiar with, mainly examine and approve the difference of standard, the requirement of medicine is higher than health product.
In the application of a first aspect of the present invention, mammal can be people, domestic animal, house pet etc., preferably people.Preferably wherein, mammal is boar, as man.
In this article, " natural " refers to and extracts from natural origin (as pollen, Cera Flava etc.) from limited material.In the specific embodiment of the present invention, the natural brassinolide analogue provided is from Pollen Brassicae campestris.Certainly, owing to having resolved these natural natural brassinolide analogue, therefore they have been not limited to extract from natural origin (as pollen, Cera Flava etc.), also can be chemosynthesis.
In the application of a first aspect of the present invention, the brassinolide analog that brassinolide analog is preferably separated.In this article, " separation " refer to brassinolide analog and depart from or once departed from naturally occurring environment and exist with the purity higher than 50% or existed.Although brassinolide analog again can be mixed in mixture thus to reduce the purity of itself, but high-purity once, be convenient to each batch products is stablized, this relates to the field of personal safety for preparing medicine or health product etc., be particularly important and than the advantage wanted.Therefore, in the application of a first aspect of the present invention, brassinolide analog is preferably the stable brassinolide analog of proportioning.In this article, " proportioning is stablized " refers to the composition of brassinolide analog in different batches product and content is identical.Because brassinolide analog can be separated by the extracting method of first aspect present invention, therefore can be used alone or proportioning use again, can accomplish that proportioning is stablized completely.
Preferably in the application of a first aspect of the present invention, described disease is prostatosis, is more preferably prostatoplasia diseases.This disease especially perplexs middle-aging male.Correspondingly, also preferred in the application of a first aspect of the present invention, described situation is prostate condition, preferably prostatic hyperplasia situation.
Through the present inventor's further investigation, find that there is many brassinolide analog and can be used in mammal medicine or health product, but and not all.Preferably in the application of a first aspect of the present invention, described brassinolide analog is not such as formula the brassinolide analog shown in BR5:
More preferably in the application of a first aspect of the present invention, described brassinolide analog be separated such as formula the brassinolide analog shown in BR6, BR1, BR2, BR3 and/or BR4 or their mixture several arbitrarily:
Most preferably in the application of a first aspect of the present invention, described brassinolide analog be separated such as formula the brassinolide analog shown in BR3:
In second aspect, the invention provides the pharmaceutical composition for prevention or treatment mammal (especially boar) disease, it comprises brassinolide analog, and pharmaceutically acceptable carrier; In addition, present invention provides the Halth-care composition for improving mammalian condition, it comprises brassinolide analog, and acceptable carrier on health product.
In this article, on pharmaceutically acceptable carrier and health product, acceptable carrier has medicine and the implication known by field of health care products technical staff respectively, refer to respectively not affect (even can strengthen) active ingredient exerts effect, to people or the nontoxic excipient substance of other mammalian safe or health care product auxiliary material.On pharmaceutically acceptable carrier and health product, acceptable carrier often can be general, comprises excipient, solubilizing agent, filler, pH adjusting agent and/or correctives.They are applied to the adjuvant of plant impurity content than routine is much lower, and homogeneity is better.
In the compositions of a second aspect of the present invention, the brassinolide analog that brassinolide analog is preferably separated.In addition, in the compositions of a second aspect of the present invention, brassinolide analog is preferably the stable brassinolide analog of proportioning.
Preferably in the compositions of a second aspect of the present invention, described disease is prostatosis, is more preferably prostatoplasia diseases.Correspondingly, also preferred in the compositions of a second aspect of the present invention, described situation is prostate condition, preferably prostatic hyperplasia situation.
The compositions of second aspect present invention can be various preparation (dosage form), is preferably solid preparation and liquid preparation.Preparation of the present invention can be unit dosage form, as tablet, pill, capsule (comprise sustained release or releasing pattern is released in delay), powder, suspensoid, granule, tincture, syrup, emulsion agent, suspension, injection, etc. dosage form and various slow release formulation, thus be applicable to various administering mode, such as oral, parenteral injection, mucosa, muscle, intravenous, subcutaneous, ophthalmic, Intradermal or the form of medication through skin etc.The compositions of preferred a second aspect of the present invention is oral formulations.
According to the deep research of the present inventor, preferably in the compositions of a second aspect of the present invention, described brassinolide analog is not such as formula the brassinolide analog shown in BR5.
More preferably in the compositions of a second aspect of the present invention, described brassinolide analog be separated such as formula the brassinolide analog shown in BR6, BR1, BR2, BR3 and/or BR4 or their mixture several arbitrarily.
Most preferably in the compositions of a second aspect of the present invention, described brassinolide analog be separated such as formula the brassinolide analog shown in BR3.
In the third aspect, the invention provides preparation and comprise the mammal medicine of brassinolide analog or the method for health product, it comprises,
(1) extract broken Pollen Brassicae campestris with 80 ~ 100% (V/V) ethanol water, after solid-liquid separation, retain filtrate (optionally wherein filtrate concentrates further), obtain alcohol dissolubility extracting solution;
(2) alcohol dissolubility extracting solution is mixed with 0 ~ 60% (V/V) ethanol water, then add extraction into ethyl acetate, retain ethyl acetate layer and add esterase and carry out incomplete reaction, then dry, obtain lipophilic extracts;
(3) lipophilic extracts is splined on silica gel chromatographic column, with the mixed liquid eluting of methanol and ethyl acetate, collects the eluent containing brassinolide analog, is dryly also dissolved in methanol, obtains silica column purification liquid;
(4) silica column purification liquid is splined on C18 reversed phase chromatographic column, with the mixed liquid eluting of acetonitrile and water, collect containing the eluent such as formula the brassinolide analog shown in BR6, BR1, BR2, BR3 and/or BR4 respectively, but do not collect containing the eluent such as formula the brassinolide analog shown in BR5; With
(5) by after eluent drying, mix with acceptable carrier on pharmaceutically acceptable carrier or health product.
The brassinolide analog such as BR6, BR1, BR2, BR3 and/or BR4 can by the extracting method eluting successively described in the specific embodiment of the present invention, and its qualification collection of illustrative plates is respectively as shown in Fig. 2 ~ Fig. 5, Fig. 7.In the specific embodiment of the present invention, BR3 or the eluent containing BR3 are most preferably collected.
In the method for third aspect present invention, in step (1), 100% (V/V) ethanol water can be used (namely, straight alcohol), but the concentration of preferred alcohol aqueous solution is 85 ~ 98% (V/V), be preferably 90 ~ 97% (V/V), be more preferably 93 ~ 96% (V/V), most preferably be 95% (V/V).
In the method for third aspect present invention, in step (1), the mode of broken Pollen Brassicae campestris is generally use physics crumbling method, comprise ultrasonication, mill broken etc.
In the method for third aspect present invention, in step (1), the ratio of Pollen Brassicae campestris and ethanol water can be optimized, and ratio is too low, will use excess ethyl alcohol, needs the concentrated of more high cost; Ratio is too high, then extract insufficient.Study through the present inventor, preferably in step (1), Pollen Brassicae campestris: the w/v of ethanol water is 50 ~ 200 grams: 200 ~ 500mL, is preferably 80 ~ 150 grams: 250 ~ 450mL, be more preferably 90 ~ 120 grams: 280 ~ 350mL, most preferably be 100 grams: 300mL.
In step (1), in order to extract completely, the filtering residue after extraction can be extracted further, and so repeatedly extracting filtering residue, merging filtrate.Therefore preferred in the method for third aspect present invention, step (1) comprises further, the filtering residue obtained after solid-liquid separation 80 ~ 100% (V/V) ethanol water extracts, then carry out solid-liquid separation, retain filtrate and merge with the middle filtrate obtained of step (1).This step other extraction steps of step (1) can adopt identical condition, and the filtering residue obtained after this step solid-liquid separation can adopt the step identical with this step to extract 0 ~ 5 time further, as long as finally all filtrate merged.
In step (1), the filtrate of filtrate or merging, volume is excessive, then the extraction efficiency of step after being unfavorable for.Therefore preferred in the method for third aspect present invention, in step (1), filtrate concentrates further.Concentrated is that drying under reduced pressure concentrates, and is preferably with the vacuum drying of 65 ~ 80 DEG C and 0.08 ~ 0.09Mpa concentrated, most preferably is with the vacuum drying of 75 DEG C and 0.085Mpa concentrated.After concentrated, preferred alcohols dissolubility extracting solution: the volume ratio of the ethanol water in step (2) is 0.5 ~ 2: 1 ~ 3, is preferably 0.8 ~ 1.5: 1.5 ~ 2.5, most preferably is 1: 2.
In the method for third aspect present invention, in step (2), 0% (V/V) ethanol water can be used (namely, pure water), but the concentration of preferred alcohol aqueous solution is 30 ~ 55% (V/V), be preferably 40 ~ 53% (V/V), be more preferably 45 ~ 52% (V/V), most preferably be 50% (V/V).
In the method for third aspect present invention, in step (2), the amount of the ethyl acetate added can be optimized.Through the present inventor's research, preferably in step (2), ethanol water: the volume ratio of ethyl acetate is 1 ~ 3: 3 ~ 8, is preferably 1.5 ~ 2.5: 4 ~ 6, most preferably is 2: 5.
In order to extract completely, the non-ethyl acetate layer (that is, after being separated ethyl acetate layer remaining layer) after extraction can be extracted further, and so repeatedly extracting non-ethyl acetate layer, combined ethyl acetate layer.Therefore preferred in the method for third aspect present invention, step (2) comprises further, and the non-ethyl acetate layer obtained after extraction adds extraction into ethyl acetate, retains ethyl acetate layer and merges with the middle ethyl acetate layer obtained of step (2).Extraction can carry out 1 ~ 8 time, is preferably 2 ~ 5 times.
Preferably in the method for third aspect present invention, in step (2), esterase, also claims lipase, hydrolyzing triglyceride or fatty acid ester can produce list or double glyceride and free fatty.Such enzyme itself is in field of food extensive use.Lipase preferably extracts the esterase from antibacterial, as being the lipase can buying acquisition from market, also can be directly from antibacterial (as, pseudomonas fluorescens, preferably CGMCC No.1.867 (i.e. AS1.867) bacterial strain) the middle esterase (as see Chinese patent application 200810046182.0) extracted.
Preferably in the method for third aspect present invention, in step (2), saccharifying enzyme, also claims glucoamylase, starch chain can be converted into glucose.Such enzyme itself is brewageing field extensive use.Saccharifying enzyme preferably extracts the saccharifying enzyme from fungus, as being can buy the saccharifying enzyme obtained from market, also can be the saccharifying enzyme directly extracted from fungus (as aspergillus niger).
In this article, " incomplete reaction " refers to and makes the substrate of enzyme reaction cessation reaction before all transforming, and keeps the multiformity of brassinolide analog thus.Usual complete reaction will carry out more than 3 hours, therefore preferred in the method for third aspect present invention, in step (2), incomplete reaction is 35 ~ 42 DEG C of reactions 0.5 ~ 2 hour, preferably 37 ~ 41 DEG C of reactions 0.75 ~ 1.5 hour, most preferably 40 DEG C of reactions 1 hour.
Preferably in the method for third aspect present invention, in step (2), drying is drying under reduced pressure, is preferably with the vacuum of 65 ~ 80 DEG C and 0.08 ~ 0.09Mpa dry, most preferably is with the vacuum of 75 DEG C and 0.085Mpa dry.
The present inventor finds, preliminary purification is advisable with silica gel chromatographic column, only needs the initial sum termination time arranging collection according to elution speed to get final product the multiple brassinolide analog of comprehensive collection, simple and easy to do.Preferably in the method for third aspect present invention, in step (3), the filler in silica gel chromatographic column is 200 ~ 300 order silica gel, is most preferably 300 order silica gel.
In step (3), the potential eluent that can select is of a great variety, and through the present inventor's research, the mixed liquid eluting of discovery methanol and ethyl acetate, product is more stable.Preferably in the method for third aspect present invention, in step (3), methanol in mixed liquid: the volume ratio of ethyl acetate is 3 ~ 8: 0.5 ~ 1.5, is preferably 4 ~ 7: 0.8 ~ 1.3, most preferably is 5: 1.
Preferably in the method for third aspect present invention, in step (3), drying is drying under reduced pressure, is preferably with the vacuum of 65 ~ 80 DEG C and 0.08 ~ 0.09Mpa dry, most preferably is with the vacuum of 75 DEG C and 0.085Mpa dry.
In step (4), the potential eluent that can select is of a great variety, and through the present inventor's research, the mixed liquid eluting of discovery acetonitrile and water, effectively can be separated various brassinolide analog.Preferably in the method for third aspect present invention, in step (4), acetonitrile in mixed liquid: the volume ratio of water is 60 ~ 90: 10 ~ 40, is preferably 70 ~ 80: 20 ~ 30, most preferably is 75: 25.
It is highly purified for extracting by the step in the method for third aspect present invention the brassinolide analog obtained.Preferably in the method for third aspect present invention, in step (4), the purity such as formula the brassinolide analog shown in BR6, BR1, BR2, BR3, BR4 and/or BR5 is greater than 90%, is preferably greater than 95%, more preferably greater than 98%, be most preferably greater than 99%.
The excellent effect that the present invention obtains is, once extracts, and finally can collect the highly purified brassinolide analog of multiple separation, cost of thus having made thinner; Extract (purification) flow process to stablize, the brassinolide analog of multiple separation can be used separately, also can proportioning is mixed uses, constant product quality and easily controlling, and reaches the requirement of people's medicine or health product; Break through brassin in prior art and be used for the restriction of plant, widen the purposes of its more high added value, can be used in treatment or prevention mammalian diseases, especially prostate (hypertrophy) disease, or can be used in improving mammalian condition, especially prostate (hypertrophy) situation.
For the ease of understanding, by by concrete drawings and Examples, the present invention is described in detail below.It is important to note that instantiation and accompanying drawing are only to illustrate, do not form limitation of the scope of the invention.Obvious those of ordinary skill in the art according to illustrating, can make various correction and change to the present invention herein within the scope of the invention, and these are revised and change and also include in scope of the present invention.In addition, the present invention refer to open source literature, and these documents are also to more clearly describe the present invention, and their entire contents is all included the present invention in and carried out reference, just looks like that repeated description is excessively the same in the description of the present invention for their full text.
Accompanying drawing explanation
Fig. 1 shows the purification collection of illustrative plates of brassinolide analog.
Fig. 2 ~ 7 show the Mass Spectrometric Identification collection of illustrative plates of each brassinolide analog, and wherein, as shown in Figure 2, as shown in Figure 3, as shown in Figure 4, as shown in Figure 5, as shown in Figure 6, BR6 as shown in Figure 7 for BR5 for BR4 for BR3 for BR2 for the mass-spectrogram of BR1.
Detailed description of the invention
The extraction of embodiment 1 brassinolide analog and qualification
Get Pollen Brassicae campestris 100 grams, add 95% (V/V) ethanol 300mL, ultrasonication, filter, retain filtrate; Filtering residue adds 95% (V/V) ethanol 300mL, ultrasonication, filters, and retains filtrate.Merging filtrate, in 75 DEG C, to be concentrated into volume be 100mL for the vacuum drying of 0.085Mpa, obtains pollen alcohol extract.
Be added in pollen alcohol extract by 50% (V/V) ethanol 200mL, mix homogeneously, then adds 500mL extraction into ethyl acetate, retains upper strata (ethyl acetate layer); Lower floor continues to add 500mL extraction into ethyl acetate, retains upper strata (ethyl acetate layer).Combined ethyl acetate layer, adds 300mL and concentrates enzyme liquid (2500U/L; See Chinese patent application 200810046182.0) process 1 hour in 40 DEG C of stirrings (45rpm), then in 75 DEG C, the vacuum of 0.085Mpa is dry, obtains lipophilic extracts.
Get lipophilic extracts and be splined on silica gel chromatographic column (2.6cm × 40cm, 300 order silica gel), add mobile phase (namely, the mixed liquid of methanol and ethyl acetate, methanol: the volume ratio of ethyl acetate is 5: 1) eluting, coutroi velocity is 4ml/ minute, collect the eluent flowed out for the 40th minute to the 150th minute, after merging in 75 DEG C, the vacuum of 0.085Mpa is completely dry, be dissolved in 10mL methanol, obtain silica column purification liquid.
Then, (column parameter is 50mm × 25cm silica column purification liquid to be splined on C18 reversed phase chromatographic column, 5 μm), add mobile phase (namely, the mixed liquid of acetonitrile and water, acetonitrile: the volume ratio of water is 75: 25) eluting, coutroi velocity is 10ml/min, eluting collection of illustrative plates as shown in Figure 1, wherein designate peak and the elution time thereof of each brassinolide analog, corresponding brassinolide analog (BR6, BR1, BR2, BR3, BR4, BR5) can be collected respectively in each elution time.The good stability of said extracted flow process, overall efficiency is high, once can obtain above-mentioned 6 kinds of brassinolide analog.
Collect brassinolide analog BR1, BR2, BR3, BR4, BR5 and BR6, with high resolution mass spec checking (mass-spectrogram of BR1 as shown in Figure 2, BR2 as shown in Figure 3, BR3 as shown in Figure 4, BR4 as shown in Figure 5, BR5 as shown in Figure 6, BR6 is as shown in Figure 7) correct, each analog purity is all higher than 99%, then entrust Institute of Analysis of Sichuan University according to standard method (mass spectral analysis (GB/T6041-2002), infrared spectrum analysis (GB/T6040-2002), superconducting pulse Fourier-transform nuclear magnetic resonance spectrum (JY/T007-1996) and molecule absorption spectrophotometric analysis (GB/T9721-2006)) test, parse the following chemical constitution of each brassinolide analog:
In addition the compound of above-mentioned purification is blended into brassin analog (referred to as " BR mixes " or " BR mixture "; Wherein BR1: BR2: BR3: BR4: BR5 and the weight ratio of BR6 be 0.4: 0.4: 0.4: 0.4: 0.4: 98)
Embodiment 2 animal safety is tested
Brassinolide analog BR1, BR2, BR3, BR4, BR5 and BR6 of Example 1 preparation respectively, and BR mixture, use a small amount of anhydrous alcohol solution respectively, then adding distil water is diluted to drug level and is: 0.005mg/mL, and wherein dehydrated alcohol volumetric concentration is: 0.0125%; The dehydrated alcohol distilled water that takes a morsel in addition be mixed with volumetric concentration be the ethanol water of 0.0125% in contrast.
Test with age in days SD infant rats (male and female half and half) with age in days weanling mice (male and female half and half) and SPF level with the SPF level NIH purchased from Da Shuo bio tech ltd, Chengdu.Process of the test is: mice or rat are divided at random 8 groups of (wherein BR monomers 6 groups, 1 group, BR mixture, blank 1 group), often organize 10, each group of male and female half and half, BR are with 0.1mg/kg gavage, the isopyknic above-mentioned ethanol water of blank group gavage every day 1 time, 3 times weekly, administration 4 weeks altogether, period weighs in and observes animal physiological state.Duration of test, animal all freely ingests the general common irradiated feed of the large and small Mus of SPF level and drinking pure.
Table 1 Mouse Weight result of variations (
n=10)
Table 2 rat body weight result of variations (
n=10)
Result as shown in Tables 1 and 2, under each Measuring Time point, between each observation group, large and small Mus body weight (change) is all without significant difference, also do not observe the animal that BR respectively organizes in addition to occur relative to the abnormal physiological condition that has of blank group, it is safe for showing that BR monomer 6 groups and BR mixture use animal.
Embodiment 3 anti-prostatic hyperplasia is tested
Brassinolide analog BR1, BR2, BR3, BR4, BR5 and BR6 of Example 1 preparation respectively, use a small amount of anhydrous alcohol solution respectively, then adding distil water is diluted to drug level and is: 0.005mg/mL, and wherein dehydrated alcohol volumetric concentration is: 0.0125%;
It is that the ethanol water of 0.0125% is as blank that the dehydrated alcohol distilled water that takes a morsel is mixed with volumetric concentration;
Separately get paracetamol (trade name: QIANLIEKANG, manufacturing enterprise: Kang Enbei Zhejiang Pharmaceutical Co, lot number: the accurate word Z33020303 of traditional Chinese medicines, specification: 0.57g/ sheet), last through pulverizing, become concentration to be with distilled water diluting: 1/60mL, as positive control.
Test with the SPF level NIH male mice (buying body weight 18-22g) purchased from Da Shuo bio tech ltd, Chengdu, duration of test, animal all freely ingests the general common irradiated feed of SPF level mice and drinking pure.
Process of the test is:
Random selecting 6 mices as modeling matched group, not injectable drug, and only with distilled water subcutaneous injection (10mL/Kg), 1 times/day, amount to 3 weeks; Remaining animal all carries out modeling, and random sub-cage rearing also with Testosterone Propionate subcutaneous injection modeling (5mg/Kg), 1 times/day, amounts to 3 weeks.After last modeling administration 24 hours, in the animal of modeling random selecting 6 only with modeling matched group, put to death after weighing respectively, get prostate and claim weight in wet base, calculate the prostate index (prostate index=prostate weight in wet base (mg)/Mouse Weight (g)) of two groups.Result shows, and the mouse prostate index (mg/g) of modeling is 0.90 ± 0.29, is significantly higher than modeling matched group (its prostate index (mg/g) is 0.57 ± 0.17).This shows, the animal of modeling there occurs prostatic hyperplasia.
Then, the mice of 80 of remainder successful modelings is divided at random 8 groups of (wherein BR monomers 6 groups, positive control 1 group, blank 1 group), often organize 10, BR each sample group is with 0.1mg/kg gavage every day 1 time, and blank group gavage every day equal-volume is as the ethanol water 1 time of blank, positive controls, with paracetamol solution gavage every day 1 time (10mL/Kg), is total to administration 20 days.Weigh after 24 hours in last administration and put to death mice, getting prostate and claim weight in wet base, calculating prostate index.
Prostate index after the medication of table 3 prostatic hyperplasia mice
Result shows, and successful mice is treated respectively through BR1, BR2, BR3, BR4 and BR6 in prostatic hyperplasia modeling, can both reduce prostate index, be used for the treatment of prostatic hyperplasia, wherein especially best with BR3 effect; But BR5, relative to blank, can not significantly reduce prostate index.
Claims (1)
1. brassinolide analog for the preparation of prevention or treatment boar prostatoplasia diseases medicine or for improve boar prostatic hyperplasia situation health product in application, wherein said brassinolide analog be separated such as formula the brassinolide analog shown in BR6
BR6。
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