CN106432391A - Novel steroid compound as well as preparation method and application thereof, and drug compound and application thereof - Google Patents
Novel steroid compound as well as preparation method and application thereof, and drug compound and application thereof Download PDFInfo
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- C07—ORGANIC CHEMISTRY
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- C07J11/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, not substituted in position 3
Abstract
The invention discloses a novel steroid compound as well as a preparation method and medical application thereof and provides a compound structure, a drug compound containing the compound structure, as well as a preparation method and application thereof. The compound is reported for the first time and is a steroid compound with a novel structure. The steroid compound can be acquired by extracting, separating and purifying from dry roots of acanthopanax. An in vitro test proves that the compound can induce obvious cycle arrest and apoptosis after the compound acts on Molt4 cells, and the compound can keep the cell cycle in the S period so as to induce apoptosis and restrain cell proliferation. The compound has a potential application value in the aspect of treating acute T lymphocytic leukemia, and can be used for developing a drug for treating the acute T lymphocytic leukemia.
Description
Technical field
The invention belongs to technical field of pharmaceuticals, and in particular to from the dry root of Radix Et Caulis Acanthopanacis Senticosi, isolated one kind has and controls
Treat steroid compound of Pancytopenia effect and preparation method thereof.
Background technology
The root and rhizome that dries of Radix Et Caulis Acanthopanacis Senticosi Acanthopanax senticosu (Rupr.Et Maxim) Harms is used as medicine, its
Stem, leaf are also pharmaceutically acceptable.Radix Et Caulis Acanthopanacis Senticosi belongs to Araliaceae with Radix Ginseng, is a kind of deciduous plant, obtains because of the close raw acupuncture of sprig
Name.Main product is in the ground such as northeast China (Heilungkiang, Jilin, Liaoning, Hebei), Far-east Area of Russia and Hokkaido, Japan, thorn
Slender acanthopanax is the typical medicinal plants in the Northeast of China, acrid in the mouth, slight bitter, warm in nature, nontoxic.Radix Et Caulis Acanthopanacis Senticosi has replenishing QI to invigorate the spleen, the kidney invigorating
Calm the nerves, beneficial able-bodied bone the effect of.Compendium of Materia Medica Radix Et Caulis Acanthopanacis Senticosi is called " this is through top grade ", can " invigorating the spleen and replenishing QI, the strong will of hard muscles and bones, long
Clothes are made light of one's life by commiting suicide resistance to old ".The saying for having " Ning get Yi expires car slender acanthopanax without gold and jade " among the people.Chinese patent medicine kind with Radix Et Caulis Acanthopanacis Senticosi as raw material
Class is various, such as Radix Et Caulis Acanthopanacis Senticosi tablet, Radix Et Caulis Acanthopanacis Senticosi injection, Anshen Bunao capsule, brain peace piece etc.；In addition tcm prescription is to Radix Et Caulis Acanthopanacis Senticosi demand
Amount is also larger, more than 5,000,000 kilograms of Radix Et Caulis Acanthopanacis Senticosi whole nation annual requirement, so that the market supply day of Radix Et Caulis Acanthopanacis Senticosi is becoming tight.
The main component of Radix Et Caulis Acanthopanacis Senticosi complex chemical composition, root and rhizome is phenol glycosides compound, and experiment proves manyprickle acanthopanax general
Glycosides is one of its bioactive ingredients.Separated go out seven kinds of Radix Et Caulis Acanthopanacis Senticosi glucoside (A, B, C, D, E, F, G) content ratio 8:30:10:12:
4:2:L, respectively daucosterol (A), Syringa oblata Lindl. phenolic glycoside (B), galactite general (C), syringaresinol bioside (D), this
There are outward E, F and G, and L.Research to stem and leaf of Radix Et Caulis Acanthopanacis Senticosi deepened continuously in recent years, and from leaf, isolated oleanolic acid is
Folium Acanthopanacis Radiciss glycosides I, K, L and M etc. of aglucon.
Modern pharmacological research show Radix Et Caulis Acanthopanacis Senticosi can enhancing body nonspecific defense ability, except with immunomodulating, anti-swollen
Tumor, defying age, radioprotective, damage-retardation are injured outside antifatigue effect, can also treat cardiovascular and cerebrovascular disease, diabetes, neurasthenia etc.
Disease.In recent years, the research in terms of passing through to Radix Et Caulis Acanthopanacis Senticosi physiologically active and chemical composition, shows Radix Et Caulis Acanthopanacis Senticosi effective ingredient for glycoside,
Wherein Syringin, E are principle active component.
Content of the invention
It is an object of the invention to provide isolated one kind a kind of dry root from Radix Et Caulis Acanthopanacis Senticosi has the acute T for the treatment of drenching
Steroid compound of bar chronic myeloid leukemia effect and preparation method thereof.
The above-mentioned purpose of the present invention is achieved by techniques below scheme：
A kind of new steroid compound, with following structural formula,
The preparation method of described compound, comprising following operating procedure：A the dry root of Radix Et Caulis Acanthopanacis Senticosi is crushed by (), use 75
～85% alcohol heat reflux is extracted, and united extraction liquid is concentrated into no alcohol taste, successively with petroleum ether, ethyl acetate and water saturated
N-butanol extraction, respectively obtains petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract；Acetic acid in (b) step (a)
The macroporous resin remove impurity of ethyl ester extract, first with 15% ethanol elution, 8 column volumes, then with 75% ethanol elution, 10 cylinders
Product, collects 75% ethanol elution, and concentrating under reduced pressure obtains 75% ethanol elution thing extractum；75% ethanol elution leaching in (c) step (b)
Cream is separated with purification on normal-phase silica gel, and it is 85 to use volume ratio successively:1、45:1、20:1、10:1 and 1:1 methylene chloride-methanol gradient is washed
Take off and obtain 5 components；D in () step (c), component 3 is separated further with purification on normal-phase silica gel, it is 25 to use volume ratio successively:1、20:1 He
10:1 methylene chloride-methanol gradient elution obtains 3 components；E in () step (d), the octadecylsilane of component 2 is bonded
Reverse phase silica gel is separated, and with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 75%, collects 8～10 column volume eluting
Liquid, eluent is concentrated under reduced pressure to give pure compound.
Further, in step (a), extracted with 80% alcohol heat reflux, united extraction liquid.
Further, the macroporous resin is AB-8 type macroporous adsorbent resin.
A kind of pharmaceutical composition, wherein containing the compound described in therapeutically effective amount and pharmaceutically acceptable carrier.
Application of the described compound in the medicine for preparing treatment Pancytopenia.
Application of the described pharmaceutical composition in the medicine for preparing treatment Pancytopenia.
When the compounds of this invention is as medicine, can directly use, or be used in the form of pharmaceutical composition.
The pharmaceutical composition contains the compounds of this invention of therapeutically effective amount, remaining for pharmaceutically acceptable, to people
Nontoxic with animal and inert pharmaceutical acceptable carrier and/or excipient.
Described pharmaceutical acceptable carrier or excipient are one or more selected from solid, semi-solid and liquid diluent, filler
And pharmaceutical preparation adjuvant.The pharmaceutical composition of the present invention is used in the form of per weight dose.Medicine of the present invention can
Patient in need for the treatment of is applied to by the form being administered orally or inject.During for being administered orally, tablet, slow releasing tablet, control can be made into
Release piece, capsule, drop pill, micropill, suspensoid, Emulsion, powder or granule, oral liquid etc.；During for injecting, sterilizing is can be made into
Aqueouss or oily solution, aseptic powder injection, liposome or Emulsion etc..
Description of the drawings
Fig. 1 is a kind of structural formula of new steroid compound；
Fig. 2 is that a kind of theoretical ECD value of new steroid compound is compared with experiment ECD value.
Specific embodiment
The essentiality content of the present invention is further illustrated with reference to embodiment, but the present invention is not limited with this and protected model
Enclose.Although being explained in detail to the present invention with reference to preferred embodiment, it will be understood by those within the art that, can be right
Technical scheme is modified or equivalent, without deviating from the spirit and scope of technical solution of the present invention.
Embodiment 1：A kind of separation preparation of new steroid compound and structural identification
Reagent source：Ethanol, petroleum ether, ethyl acetate, n-butyl alcohol, dichloromethane are pure for analysis, insult peaking purchased from Shanghai
Reagent company limited, methanol, analysis is pure, purchased from Jiangsu Han Bang chemical reagent company limited.
Preparation method：A the dry root (10kg) of () Radix Et Caulis Acanthopanacis Senticosi is crushed, extract (25L × 3 time) with 80% alcohol heat reflux,
United extraction liquid, is concentrated into no alcohol taste (3L), successively with petroleum ether (3L × 3 time), ethyl acetate (3L × 3 time) and water saturated
N-butyl alcohol (3L × 3 time) is extracted, and respectively obtains petroleum ether extract, acetic acid ethyl ester extract (398g) and n-butyl alcohol extract；
B acetic acid ethyl ester extract AB-8 type macroporous resin remove impurity in () step (a), first with 15% ethanol elution, 8 column volumes, then uses
75% column volume of ethanol elution 10, collects 75% ethanol elution, and concentrating under reduced pressure obtains 75% ethanol elution thing extractum (163g)；
C in () step (b), 75% ethanol elution extractum is separated with 200-300 mesh purification on normal-phase silica gel, it is 85 to use volume ratio successively:1 (10 posts
Volume), 45:1 (9 column volumes), 20:1 (8 column volumes), 10:1 (8 column volumes) and 1:The dichloromethane of 1 (5 column volumes)
Alkane-methanol elution gradient obtains 5 components；D in () step (c), component 3 (49g) is divided further with 200-300 mesh purification on normal-phase silica gel
From it is 25 to use volume ratio successively:1 (8 column volumes), 20:1 (10 column volumes) and 10:The dichloromethane of 1 (5 column volumes)-
Methanol elution gradient obtains 3 components；E reverse phase silica gel that in () step (d), component 2 (31g) is bonded with octadecylsilane
ODS-C18 is separated, and with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 75%, collects 8～10 column volume eluents,
Eluent is concentrated under reduced pressure to give pure compound (244mg).
Structural identification：White powder；HR-ESIMS shows [M+Na]+For m/z 449.3014, can obtain in conjunction with nuclear-magnetism feature
Molecular formula is C28H42O3, degree of unsaturation is 8.Hydrogen nuclear magnetic resonance modal data δH(ppm, DMSO-d6, 600MHz)：H-1 (1.74,
M), H-1 (2.01, m), H-2 (5.68, m), H-3 (5.67, m), H-4 (1.99, m), H-4 (2.25, m), H-6 (5.91, d, J=
10.0), H-7 (5.12, d, J=10.0), H-11 (1.38, m), H-11 (1.89, m), H-12 (1.35, m), H-12 (1.88,
M), H-15 (1.43, m), H-15 (1.76, m), H-16 (1.38, m), H-17 (1.49, m), H-18 (0.89, s), H-19
(1.07, s), H-20 (2.01, m), H-21 (0.98, d, J=6.5), H-22 (5.13, dd, J=15.3,7.7), H-23
(5.16, dd, J=15.3,7.0), and H-24 (1.78, m), H-25 (1.37, m), H-26 (0.79, d, J=7.0), H-27
(0.82, d, J=7.0), H-28 (0.88, d, J=6.6)；Carbon-13 nmr spectra data δC(ppm, DMSO-d6, 150MHz)：
29.7(CH2, 1-C), 123.8 (CH, 2-C), 126.4 (CH, 3-C), 36.5 (CH2, 4-C), 72.6 (C, 5-C), 140.3 (CH,
6-C), 123.8 (CH, 7-C), 67.1 (C, 8-C), 75.2 (C, 9-C), 41.7 (C, 10-C), 25.3 (CH2, 11-C), 27.7
(CH2, 12-C), 41.8 (C, 13-C), 83.6 (C, 14-C), 28.1 (CH2, 15-C), 26.3 (CH2, 16-C), 51.6 (CH, 17-
C), 15.2 (CH3, 18-C), 21.2 (CH3, 19-C), 39.1 (CH, 20-C), 20.7 (CH3, 21-C), 134.3 (CH, 22-C),
(132.5 CH, 23-C), 42.4 (CH, 24-C), 32.7 (CH, 25-C), 19.6 (CH3, 26-C), 20.1 (CH3, 27-C), 17.5
(CH3, 28-C)；Carbon atom labelling is referring to Fig. 1.1H H NMR spectroscopy shows six methyl signals [δ H0.79 (d, J=7.0Hz, Me-
26), 0.82 (d, J=7.0Hz, Me-27), 0.88 (d, J=6.6Hz, Me-28), 0.89 (s, Me-18), 0.98 (d, J=
6.5Hz, Me-21), 1.07 (s, Me-19)], six olefinic methine proton signals [δ H5.12 (d, J=10.0Hz, H-7),
5.13 (dd, J=15.3,7.7Hz, H-22), 5.16 (dd, J=15.3,7.0Hz, H-23), 5.68 (m, H-2), 5.67 (m, H-
3), 5.91 (d, J=10.0Hz, H-6)].13C H NMR spectroscopy shows 28 resonance carbon signals, including six methyl signals [δ C15.2
(C-18), 17.5 (C-28), 19.6 (C-26), 20.1 (C-27), 20.7 (C-21), 21.2 (C-19)], six methylene, ten
Individual methine [six olefinics methine δ C123.8 (C-2), 123.8 (C-7), 126.4 (C-3), 132.5 (C-23), 134.3
(C-22), 140.3 (C-6)], and six quaternary carbons [four oxygen-containing quaternary carbon δ C67.1 (C-8), 72.6 (C-5), 75.2 (C-9), 83.6
(C-14)].In HMBC spectrum, H2- 4 (δ H1.99/2.25) and C-5 (δ C72.6) and C-6 (δ C140.3)；Me-19 (δ H1.07) with
C-5 (δ C72.6) and C-9 (δ C75.2)；H-6 (δ H5.91) and C-8 (δ C67.1) and C-5 (δ C72.6)；H-7 (δ H5.12) with
C-8 (δ C67.1), C-9 (δ C75.2) and C-14 (δ C83.6)；And the dependency of Me-18 (δ H0.89) and C-14 (δ C83.6)
Show that C-5 and C-14 position is respectively connected with a hydroxyl, between C-8 and C-9, there is epoxide group.NOESY spectrum shows B ring for boat form structure
Type, then 8,9- epoxide group is α configuration.Between H-22 and H-23, big coupling constant (15.3Hz) shows between C-22 and C-23
Double bond be E.Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrum and NOESY spectrum, and document is with regard to correlation type nuclear magnetic data, can base
This determines the compound as shown in figure 1, spatial configuration determines, theoretical value is basically identical with experiment value by ECD test further
(Fig. 2).
Embodiment 2：A kind of pharmacological action test of new steroid compound
First, material and instrument
Pancytopenia Molt4 cell is given by Ji'nan University's hematopathy institute；The white blood of chronic granulocyte
Sick cell K562 cell is given by Biochemistry and Molecular Biology teaching and research room by Ji'nan University.Compound is returned for self-control, HPLC
One changes purity is more than 98%, is configured to 5.5 μ g/mL solution for standby with the pure dissolving of DMSO analysis.DMEM/F12 medium powder, hydroxyl second
Base piperazine ethanesulfonic acid (HEPES), tetraphenyl nitrogen indigo plant reagent (MTT) are purchased from Gibeo company of the U.S..New-born calf serum (Newbom
Calf Serum) it is purchased from Hangzhou China Ilex purpurea Hassk.[I.chinensis Sims company.Penicillin (Penicillin), streptomycin (Streptomycin) are purchased from
China of North China drugmaker.The third eyelash of iodate (PT) is purchased from Sigma Co., USA.The double transfection reagent boxes of PI-Annexin V are purchased from China
Beijing Bao Sai biotech company.The high-quality fluorimetric reagent box of ROS is purchased from Chinese green skies biotechnology research institute.
Rhodamine123 is purchased from Molecular Probes company of the U.S..
CO2Incubator (U.S. Thermo Forma), superclean bench (Chinese Suzhou purifying apparatus factory), steam is gone out
Bacterium pot (LDZX40BI) (Chinese Shanghai Shen An medical apparatus and instruments factory), TGL-16G type table model high speed centrifuge (Chinese Shanghai An Ting section
Learn instrument plant), MA260S type electronic analytical balance (the second balance equipment of Chinese Shanghai factory), automatically dual pure water distillator (in
One factory of state's Shanghai glass apparatus), Sterivex TM, 0.22 μm of Millex Syringe Filters (Millipore company of the U.S.),
XDS-1B optics inverted microscope (Chongqing in China optical instrument factory), full-automatic microplate reader (BIO-RID company of the U.S.), FACS
Calibur flow cytometer (BD FACSAria company of the U.S.).(kylin is medical for Jiangsu Haimen City for QL-901 type vortex mixer
Instrument plant).
2nd, test method
1st, cell culture
People's Pancytopenia cell Molt4 cell culture system：DMEM/F12 culture fluid contains 10% new born bovine
Serum, put 37 DEG C, volume fraction be 5%CO2Incubator, per 2-3 days Secondary Culture.From exponential phase, 0.2% trypan blue
Refuse dye rate>95% cell is tested.
2nd, mtt assay detection cell proliferation inhibition rate
3×104ML Molt4 cell is inoculated in 96 well culture plates, and final volume is 200 μ L/ holes, to set 5 multiple holes, chemical combination per group
Thing sets five concentration and is respectively 2.5 μ g/mL, 5 μ g/mL, 10 μ g/mL, 15 μ g//mL, 20 μ g//mL.Put 37 DEG C, 5%CO2Culture
In case after culture 48h, 72h, MTT solution (5mg/mL PBS is added per hole<Ph=7.4 joins) 20 μ L, continue incubation 4 hours, eventually
Only cultivate, centrifugation, careful suction abandons in the hole culture supernatant, adds 150 μ L DMSO per hole, vibrates 10 minutes, so that crystal is fully melted
Solution, microplate reader detection light absorption value (determining wavelength 570nm, reference wavelength 690nm), calculate each group proliferation inhibition rate.Test every time
It is repeated 3 times, takes its average.Proliferation inhibition rate (%)=(1-A experimental group/A matched group) × 100%.
3rd, the mono- dye detection cell cycle of PI
2×l05/ mL Molt4 cell is inoculated in 12 well culture plates, and per 1800 μ L of hole, compound (I) sets three concentration and divides
Not Wei 15 μ g/mL of 5 μ g/mL, 10 μ g/mL, 2500 μ L of final volume, if 3 multiple holes.Put 37 DEG C, 5%CO248h is cultivated in incubator
Afterwards collect cell, the 0.01mol/L PBS washed cell of pre-cooling 2 times, with volume fraction be 70% ethanol, 4 DEG C of fixations overnight, from
The heart removes supernatant, adds PI dyeing liquor (containing RNase), 50 μ g/mL of final concentration, and lucifuge dyes 30min, after 300 mesh nylon net filters
Flow cytometer analysis cell DNA content, 12000 cells of each sample stochastic analysis, obtain each group cell growth cycle ratio
Example, U.S.'s BD FAC Sort Cell Quest software analysis result.
4th, the double dyes of AnnexinV-PI determine apoptosis ratio
All ibid, after culture 48h, adjustment cell concentration is that 5 × 106/mL, each group takes lmL for cell process and adding method thereof
Cell, pre-cooling 0.01mol/L PBS is washed 3 times, exhausts supernatant, adds the combination buffer of 200 μ L test kits offer, resuspended thin
Born of the same parents, are separately added into 10 μ L Annexin V-FITC and 5 μ L, gently mix, and 4 DEG C of lucifuges react 30min, add 300 μ L combination
Buffer, flow cytometer is detected.The cell mass (i.e. LR cell mass) of Annexin V-FITC+, PI- is that early apoptosis are thin
Born of the same parents.
5th, statistical analysiss
With mean scholar's standard deviationRepresent, the process of application SPSS13.0 statistical software, using completely randomized design
The significance of one factor analysis of variance (one-wayANOVA) analysis group difference.
3rd, result and conclusion
1st, inhibited proliferation of the compound to Molt4 cell
Compound all has obvious inhibited proliferation in different time points to Molt4 cell.DMSO (0.5%) group exists
The proliferation inhibition rate of 48h, 72h is 5.2% ± 0.8%, 6.40% ± 0.9% respectively.The increasing of variable concentrations compound treatment group
Grow suppression ratio all statistically significant (P compared with DMSO group<0.01).Under the conditions of same concentrations, the proliferation inhibition rate of 72h is relatively
48h height.IC of the compound to Molt4 cell 48h, 72h50It is 19.4 ± 0.2 μ g/mL, 15.7 ± 0.1 μ g/mL respectively.Different dense
The compound of degree all shows certain proliferation inhibiting effect, and relies relation with time, dosage.The results are shown in Table 1 (note：* labelling with
Control group compares, P<0.01).
2nd, impact of the compound to Molt4 cell cycle
After compound effects Molt4 cell 48h, blank control group G1 phase cell proportion is that 37.5% ± 0.2%, the S phase is thin
It is 39.6% for 11.7% ± 0.1%, DMSO group G1 phase cell proportion that born of the same parents' ratio is 50.6% ± 0.1%, G2 phase cell proportion
It is 8.7% ± 0.2% that ± 0.1%, S phase cell proportion is 51.6% ± 0.2%, G2 phase cell proportion.20 μ g/ of compound medicine
ML, cell proportion shared by 25 μ g/mL treatment group G1 phases are respectively shared by 4.1% ± 0.1%, 65.8% ± 0.1%, S phase cell
Ratio increase be respectively 85.1% ± 0.3%, 87.1% ± 0.25 (P<0.01), G2 phase cell proportion is respectively
10.7% ± 0.2%, 7.0% ± 0.1%, as a result show that there is significant difference (P the G1 phase<0.05) compound retardance Molt4 is described
Cell is in the phase in cycle S.The results are shown in Table 2 (notes：* labelling is compared with Control group, P<0.01).
3rd, the early apoptosis rate after compound effects Molt4 cell
After compound effects Molt4 cell 48h, the double dye flow cytomery early apoptosis rates of Almexin V/PI.Empty
7.0% scholar 0.3% of early apoptosis rate of white cellular control unit early apoptosis rate 6.6% ± 0.4%, DMSO matched group, with blank
Not statistically significant (p is compared in control>0.05).10 μ g/mL group of compound, 15 μ g/mL group early apoptosis rates are 9.5% respectively ±
0.3%th, 15.0% ± 0.5%, (P statistically significant compared with blank<0.05).The results are shown in Table 3 (notes：* labelling with
Control group compares, P<0.05).
Conclusion, can induce after we have found that compound effects Molt4 cell under study for action its significant Cycle Arrest and
Apoptosis, and cell cycle arrest is made in the S phase, so as to inducing cell apoptosis, suppress cell propagation.Compound is treating acute T pouring
With potential using value in terms of bar chronic myeloid leukemia.
1 compound of table to the inhibited proliferation of Molt4 cell (%,N=3)
| Group | 48h suppression ratio (%) | 72h suppression ratio (%) |
| DMSO | 5.2±0.8 | 6.4±0.9 |
| 2.5 μ g/mL of compound | 6.3±1.3 | 8.4±1.1 |
| 5 μ g/mL of compound | 10.5±2.4 | 11.6±2.5 |
| 10 μ g/mL of compound | 18.6±1.6* | 26.9±2.7* |
| 15 μ g/mL of compound | 33.4±2.7* | 49.4±2.2* |
| 20 μ g/mL of compound | 53.2±4.4* | 65.4±1.6* |
2 each group cell cycle percentage ratio of table (%,N=3)
3 compound of table on the apoptotic impact of Molt4 (%,N=3)
| Group | Apoptosis rate |
| Control | 6.6±0.2 |
| DMSO | 7.0±0.4 |
| 10 μ g/mL of compound | 9.5±0.3* |
| 15 μ g/mL of compound | 15.0±0.3* |
Embodiment 3
The preparation of tablet：Compound is first obtained by 1 method of embodiment, and utilizes organic acid such as tartaric acid or citric acid
Or the salt that formic acid or ethanedioic acid etc., mineral acid such as hydrochloric acid or sulphuric acid or phosphoric acid are made, by itself and excipient weight ratio be:9 ratio
Example adds excipient, pelletizing press sheet.
Embodiment 4
The preparation of oral liquid：Compound is first obtained by 1 method of embodiment, and utilizes organic acid such as tartaric acid or Fructus Citri Limoniae
Acid or the salt made of formic acid or ethanedioic acid etc., mineral acid such as hydrochloric acid or sulphuric acid or phosphoric acid, routinely oral liquid preparation method make oral
Liquid.
Embodiment 5
Capsule or the preparation of granule：Compound is first obtained by 1 method of embodiment, and utilizes organic acid such as winestone
The salt that acid or citric acid or formic acid or ethanedioic acid etc., mineral acid such as hydrochloric acid or sulphuric acid or phosphoric acid are made, by itself and excipient weight
Than for 1:9 ratio adds excipient, makes capsule or granule.
Embodiment 6
The preparation of injection：Compound is first obtained by 1 method of embodiment, and utilizes organic acid such as tartaric acid or Fructus Citri Limoniae
Acid or the salt made of formic acid or ethanedioic acid etc., mineral acid such as hydrochloric acid or sulphuric acid or phosphoric acid, routinely plus water for injection, fine straining, filling
Injection is made in envelope sterilizing.
Embodiment 7
The preparation of aseptic powder injection：Compound is first obtained by 1 method of embodiment, and utilizes organic acid such as tartaric acid or lemon
The salt that lemon acid or formic acid or ethanedioic acid etc., mineral acid such as hydrochloric acid or sulphuric acid or phosphoric acid are made, is dissolved in sterile water for injection,
Stirring makes molten, is filtered with aseptic suction funnel, then aseptic fine straining, is sub-packed in ampoule, and after frozen drying, aseptic sealing by fusing is obtained
Injectable powder.
The effect of above-described embodiment indicates that the essentiality content of the present invention, but does not limit the protection of the present invention with this
Scope.It will be understood by those within the art that, technical scheme can be modified or equivalent,
Essence and protection domain without deviating from technical solution of the present invention.
Claims (7)
1. a kind of new steroid compound, it is characterised in that：The compound has following structural formula,
2. the preparation method of the compound described in claim 1, it is characterised in that comprising following operating procedure：A () is by Radix Et Caulis Acanthopanacis Senticosi
Dry root crush, with 75～85% alcohol heat reflux extract, united extraction liquid, be concentrated into no alcohol taste, successively use petroleum ether, second
Acetoacetic ester and water saturated n-butanol extraction, respectively obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract；
B acetic acid ethyl ester extract macroporous resin remove impurity in () step (a), first with 15% ethanol elution, 8 column volumes, then uses 75% second
10 column volumes of alcohol eluting, collect 75% ethanol elution, and concentrating under reduced pressure obtains 75% ethanol elution thing extractum；In (c) step (b)
75% ethanol elution extractum is separated with purification on normal-phase silica gel, and it is 85 to use volume ratio successively:1、45:1、20:1、10:1 and 1:1 dichloromethane
Alkane-methanol elution gradient obtains 5 components；D in () step (c), component 3 is separated further with purification on normal-phase silica gel, use volume ratio successively
For 25:1、20:1 and 10:1 methylene chloride-methanol gradient elution obtains 3 components；E in () step (d), component 2 uses octadecane
The reverse phase silica gel of base silane bonding is separated, and with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 75%, collects 8～10
Column volume eluent, eluent is concentrated under reduced pressure to give pure compound.
3. the preparation method of compound according to claim 2, it is characterised in that：In step (a), returned with 80% ethanol heat
Stream is extracted, united extraction liquid.
4. the preparation method of compound according to claim 2, it is characterised in that：The macroporous resin is AB-8 type macropore
Adsorbent resin.
5. a kind of pharmaceutical composition, it is characterised in that：The compound described in claim 1 and medicine wherein containing therapeutically effective amount
Acceptable carrier on.
6. application of the compound described in claim 1 in the medicine for preparing treatment Pancytopenia.
7. application of the pharmaceutical composition described in claim 5 in the medicine for preparing treatment Pancytopenia.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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