CN107982315A - A kind of Chinese medicine composition for treating leukaemia and preparation method thereof - Google Patents
A kind of Chinese medicine composition for treating leukaemia and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of Chinese medicine composition for treating leukaemia and preparation method thereof, and it is preparing the application in treating acute promyelocytic leukemic medicine.The present invention improves traditional herbal medicine compound realgar natural indigo tablet, fine grinding realgar in compound realgar natural indigo tablet prescription is replaced with into realgar microorganism extracting liquid, it is big to solve fine grinding realgar dosage in compound realgar natural indigo tablet, toxicity is high, bioavilability is low, preparation is coarse, is difficult to the problems such as absorption and therapeutic effect difference, the dosage of realgar in active compound side is reduced, reduces toxicity, its therapeutic effect to acute promyelocytic leukemic is better than compound realgar natural indigo tablet.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition for treating leukaemia, and in particular to its preparation method and its prepare treat
Application in acute promyelocytic leukemia medicine, belongs to biomedicine field.
Background technology
Acute promyelocytic leukemia (APL) is one kind in leukaemia, belongs to acute non lymphocytic leukemia, mesh
Before be difficult to cure.
Chinese patent drug compound realgar natural indigo tablet (manufacturer：Anhui Province Tiankang Medicine Industry Co., Ltd, authentication code：Chinese medicines quasi-word
Z20090788, batch number：131101, major function：Acute promyelocytic leukemia) clinically it has been employed successfully in
Treat APL.But fine grinding realgar dosage is big existing for compound realgar natural indigo tablet, toxicity is high, bioavilability is low, preparation is coarse, is difficult to
Absorb and limit its medical value with problems such as therapeutic effect differences.Some scholars in order to realize the purpose of attenuation synergistic,
Suspension is made in realgar using the method for physics, chemistry mostly or is improved using nanometer technology reduction realgar grain diameter
It is dissolved, but fails to realize attenuation synergistic well.
Realgar in compound realgar natural indigo tablet is replaced with realgar microorganism extracting liquid by the present invention, solves compound realgar natural indigo tablet prescription
Middle fine grinding realgar dosage is big, toxicity is high, bioavilability is low, preparation is coarse, is difficult to the problems such as absorption and therapeutic effect difference,
The dosage of realgar in active compound side is reduced, reduces toxicity, its therapeutic effect to acute promyelocytic leukemic is yellow better than compound
Black pigment used by women in ancient times to paint their eyebrows piece.
The content of the invention
It is an object of the present invention to provide a kind of Chinese medicine composition for treating leukaemia and preparation method thereof, the present invention also has one
The Chinese medicine composition that purpose is to provide the treatment leukaemia is preparing the application in treating acute promyelocytic leukemia medicine.
The present invention is achieved through the following technical solutions.
The present invention provides a kind of Chinese medicine composition for treating leukaemia, the Chinese medicine composition includes following proportionings and is
33.5：40：12：The indigo naturalis of 10m/m/m/v, Radix Salviae Miltiorrhizae, radix pseudostellariae and realgar microorganism extracting liquid.The realgar microorganism extracting liquid
Total arsenic concentration be 461.4 μ g/ml.
Wherein, the realgar microorganism extracting liquid is prepared by following methods：Take Fe containing 1g/L2+The 0K of ferrous sulfate
Culture medium 90ml, adds realgar powder 0.5g, the Thiobacillus ferrooxidans of inoculation 10ml domestications, in 30 DEG C, pH1.8,130rpm
Constant temperature oscillation Shaking culture leaches 25-30 days, and precipitation is abandoned in filtering, is obtained filtrate, is taken the NaOH solution containing 2mol/L EDTA-2Na
It is 7.0 to adjust filtrate pH value, up to realgar microorganism extracting liquid.
Wherein, the Thiobacillus ferrooxidans is the Thiobacillus ferrooxidans of arsenic domestication, and domestication flow is：Take oxidation sub-
Iron Thiobacillus, it is sub- with 200 μ g sodium arsenites/100ml 9K culture mediums, 400 μ g sodium arsenites/100ml 9K culture mediums, 600 μ g
Natrium arsenicum/100ml 9K culture mediums, 800 μ g sodium arsenites/100ml 9K culture mediums, 1600 μ g sodium arsenites/100ml 9K trainings
The domestication of base gradient is supported, then takes 0.1g realgars/100ml 9K culture mediums, 0.2g realgars/100ml 9K culture mediums, 0.3g male again
Huang/100ml 9K culture mediums, 0.5g realgars/100ml 9K culture mediums, 1.0g realgars/100ml 9K culture mediums gradient domestication oxidation
Ferrous Thiobacillus, gradually reduces the amount of ferrous sulfate in culture medium afterwards, when the amount of ferrous sulfate is 3K, obtains domestication oxidation
Ferrous Thiobacillus.
Wherein, total arsenic concentration of the realgar microorganism extracting liquid is 461.4 μ g/ml.
Preferably, the Chinese medicine composition of the treatment leukaemia includes following raw materials：Indigo naturalis 33.5g, Radix Salviae Miltiorrhizae 40g, crown prince
Join 12g, realgar microorganism extracting liquid 10ml, the arsenic content of the arsenic content of the Chinese medicine composition equivalent to 7.3g raw material realgars.
The invention also discloses the Chinese medicine composition answering in treatment acute promyelocytic leukemia medicine is prepared
With.
In addition, the preparation method of the Chinese medicine composition the invention discloses treatment leukaemia：
(1) radix pseudostellariae, indigo naturalis are taken, is crushed, is uniformly mixed spare
(2) Radix Salviae Miltiorrhizae is taken, adds distilled water, is evaporated under reduced pressure to the filtrate of the Radix Salviae Miltiorrhizae of concentration, adds realgar microorganism leaching
Liquid, drying obtain medicinal extract；
(3) medicinal extract for obtaining step (2) adds the radix pseudostellariae indigo naturalis mixture that step (1) obtains, and dries, and crushes, and adjusts
Even, granulation, is drying to obtain the Chinese medicine composition for treating leukaemia.
The main component of Chinese medicine composition is realgar microorganism extracting liquid, indigo naturalis, Radix Salviae Miltiorrhizae.Realgar microorganism extracting liquid is place
Monarch drug in a prescription in side.Arsenic is main pharmacodynamics component.
The invention has the advantages that Chinese medicine composition not only significantly reduces the dosage of realgar, reduce toxicity, its is right
The therapeutic effect of acute promyelocytic leukemic is better than compound realgar natural indigo tablet.And detected by cell cycle, WB, said composition
Tumour cell can be made obvious apoptotic body occur, by contrasting commercially available compound realgar natural indigo tablet, when invention Chinese traditional medicine composition
When arsenic concentration is commercially available 1/2 in thing, remain able to reach identical therapeutic effect.
Brief description of the drawings
Fig. 1 is inhibition of the Chinese medicine composition to NB4 cells.
Fig. 2 is inhibition of the Chinese medicine composition to HL-60 cells.
Fig. 3, Fig. 4, Fig. 5, Fig. 6 are drug effect NB4Apoptosis result afterwards.
Fig. 7, Fig. 8, Fig. 9, Figure 10 are Apoptosis result after medicine HL-60.
RTS is realgar leachate in figure；FRBS is the multiple Chinese medicine composition of the present invention；Original formulae are water
Fly the compound realgar natural indigo tablet of realgar；RIF is compound realgar natural indigo tablet (authentication code：Chinese medicines quasi-word Z20090788), Anhui Province's Tian Kang medicine companies
Co., Ltd produces.
Embodiment
Technical scheme is described in further detail with reference to specific embodiment, but protection scope of the present invention is not
It is confined to as described below.
The preparation of 1 Chinese medicine composition tablet of embodiment
1. biomaterial
Thiobacillus ferrooxidans (Acidithiobacillus ferrooxidans BY3-1) is on July 23rd, 2004
China typical culture collection center is preserved in, deposit number is CCTCC NO:M 204057, entitled Thiobacillus ferrooxidans
(Acidithiobacillus ferrooxidans BY3-1).Preservation address is Wuhan, China Wuhan University.
2. medicine
9K fluid nutrient mediums (g/L)：(NH4)2SO43.0g, K2HPO40.5g, KCl 0.1g, MgSO4·7H2O 0.5g,
Ca(NO3)20.01g, FeSO4·7H2O 44.78g, 1L is settled to distilled water.
3K fluid nutrient mediums (g/L)：(NH4)2SO43.0g, K2HPO40.5g, KCl 0.1g, MgSO4·7H2O 0.5g,
Ca(NO3)20.01g, FeSO4·7H2O 14.93g, 1L is settled to distilled water.
0K fluid nutrient mediums (g/L)：(NH4)2SO43.0g, K2HPO40.5g, KCl 0.1g, MgSO4·7H2O 0.5g,
Ca(NO3)20.01g, 1L is settled to distilled water.
3. the preparation of realgar leachate
Thiobacillus ferrooxidans is taken, with 200 μ g sodium arsenites/100ml 9K culture mediums, 400 μ g sodium arsenites/100ml
9K culture mediums, 600 μ g sodium arsenites/100ml 9K culture mediums, 800 μ g sodium arsenites/100ml 9K culture mediums, 1600 μ g arsenous
Sour sodium/100ml 9K culture mediums gradient domestication, then takes 0.1g realgars/100ml 9K culture mediums, 0.2g realgars/100ml 9K again
Culture medium, 0.3g realgars/100ml 9K culture mediums, 0.5g realgars/100ml 9K culture mediums, 1.0g realgars/100ml 9K cultures
Base gradient is tamed, and gradually reduces the amount of ferrous sulfate in culture medium afterwards, when the amount of ferrous sulfate is 3K, obtains domestication oxidation
Ferrous Thiobacillus.
Take Fe containing 1g/L2+The 0K culture medium 90ml of ferrous sulfate, add realgar powder 0.5g, the oxygen of inoculation 10ml domestications
Change ferrous Thiobacillus, leached 25-30 days in 30 DEG C, pH1.8,130rpm constant temperature oscillation Shaking culture, precipitation is abandoned in filtering, must be filtered
Liquid, it is 7.0 to take the NaOH solution containing 2mol/L EDTA-2Na to adjust filtrate pH value, and up to realgar microorganism extracting liquid, its is dense
Spend for 461.4 μ g/ml.
4. the preparation of Chinese medicine composition tablet
(1) it is spare to prepare realgar microorganism extracting liquid 10ml；
(2) radix pseudostellariae 12g is spare after crushing；
(3) it is spare to crush indigo naturalis 33.5g；
(4) Heterophylly Falsesatarwort Root and indigo naturalis powder obtained above-mentioned steps (2), (3) is uniformly mixed spare；
(5) Radix Salviae Miltiorrhizae 40g is weighed, adds the distilled water of 480ml every time, Radix Salviae Miltiorrhizae is extracted four times, each 1.5h with distillation under vacuum,
Merge the filtrate of four times, then concentrate the filtrate of Radix Salviae Miltiorrhizae, add the realgar leachate of step (1) acquisition, in 40 DEG C or so bakings
It is 1.50-1.55 medicinal extract into relative density；
(6) medicinal extract that step (5) obtains is added in the radix pseudostellariae indigo naturalis mixture of step (4) acquisition, mixed, 50
DEG C constant temperature oven in dry, crushed, mixed well with universal high speed disintegrator FLB-200A types, pelletized, it is dry, add 1%m/m
Magnesium stearate, mixing, whole grain, tabletting (weight 0.3g).
Inhibitory action of 2 Chinese medicine composition of embodiment to NB4 and HL-60 cells
Biomaterial
(1) NB4 cells (human acute myeloid leukaemia), purchased from Shanghai Hui Ying bio tech ltd.
(2) HL-60 (the early young grain acute leukemia cells of people) is purchased from Shanghai Hui Ying bio tech ltd.
Direct sources：Shanghai Hui Ying bio tech ltd, address：508 room (the Jiang Qiao of Hexuan Road, Shanghai City Lane 26 20
No. 5 building 508 of WanDa Plaza) postcode：200081 phones：021-56980388 faxes：021-56980380
2. reagent
Realgar microorganism extracting liquid (RTS), 10 μ g/ml of arsenic content, obtain after realgar microorganism extracting liquid stoste is diluted
The realgar microorganism extracting liquid of 10 μ g/ml of arsenic content.
The Chinese medicine composition (FRBS) for the treatment of leukaemia disclosed by the invention, 20 μ g/ml of arsenic content.
The compound realgar natural indigo tablet (Original formulae) of fine grinding realgar is to be prepared according to CN100393329C methods
Original formulation medicine, 40 μ g/ml of arsenic content.
Compound realgar natural indigo tablet (RIF) (authentication code：Chinese medicines quasi-word Z20090788), purchased from the limited public affairs of Anhui Province's Tian Kang medicine companies
Department, 40 μ g/ml of arsenic content.
The preparation of three liquid：Lauryl sodium sulfate 20g is weighed, while measures isobutanol 10ml, HCl 36.5%V/V
0.2ml, above-mentioned substance is added in 190ml water, be sufficiently stirred, be uniformly dissolved after obtain three liquid 200ml, protect at room temperature
Deposit.
3. experiment and result
(1) processing of experimental drug：Above-mentioned solid medicine adds distilled water, then add the DMSO of 2%v/v to exist after weighing
4h is stirred in blender, is filtered with 0.22 μm of filter, it is stand-by.
(2) mtt assay is used, is 1 × 10 by cell concentration6Cell (the acute early young grain acute leukemia cells of people of a/ml
The NB4 or early young grain acute leukemia cells HL-60 of people) it is inoculated in 96 orifice plates, add 0.1ml per hole, 3 parallel holes are done, with 96
Orifice plate is repeated twice.37 DEG C are put, 5%CO2In incubator cultivate 24h after, add different arsenic concentrations sample solution (RTS,
FRBS, RIF, original formulae), per hole 0.1ml, 24h is overnight.Add 20 μ l of MTT, continue to cultivate 4h, then add three
150 μ l of liquid, terminate reaction overnight.Absorbance (OD) value is measured with microplate reader 570nm, cell growth inhibition is calculated by following equation
Rate (IR)：IR (%)=(1- experimental groups OD570/ control group OD570) × 100%.The result is shown in Figure 1, Fig. 2.
Experimental result is shown：In 24h, 36h and 48h, the RIF (compound realgar natural indigo tablet) and arsenic content of 40 μ g/ml of arsenic content
The FRBS (multiple Chinese medicine composition of the invention) of 20 μ g/ml has NB4 cells inhibitory action (Fig. 1), inhibiting rates of the FRBS to NB4
Higher than RIF to NB4 inhibiting rates, illustrate that apoptosis-induced effects of the FRBS to NB4 is better than apoptosis-induced effects of the RIF to NB4；And
Inhibiting rate also suppression apparently higher than original formulae (compound realgar natural indigo tablet of fine grinding realgar) to NB4 of the FRBS to NB4
Rate, this further demonstrates the FRBS containing low dose of arsenic is stronger to NB4 inhibitory action, the ability of RTS induction NB4 Apoptosis is strong
In FRBS, RIF and original formulae.
In 24h, 36h and 48h, the FRBS of the 20 μ g/ml of RIF (compound realgar natural indigo tablet) and arsenic content of 40 μ g/ml of arsenic content
(multiple Chinese medicine composition of the invention) has HL-60 cells inhibitory action (Fig. 2), and FRBS is higher than RIF pairs to the inhibiting rate of HL-60
HL-60 inhibiting rates, illustrate that apoptosis-induced effects of the FRBS to NB4 is better than apoptosis-induced effects of the RIF to HL-60；And FRBS
To the inhibiting rate of the inhibiting rate of HL-60 apparently higher than original formulae (compound realgar natural indigo tablet) to HL-60, this is further
It is stronger to HL-60 inhibitory action to demonstrate the FRBS containing low dose of arsenic.
3 Chinese medicine composition of embodiment detects the apoptosis of NB4 cells
Biomaterial, reagent are the same as embodiment two.
After FRBS, RIF processing NB4 cells 24h, cell suspension (1 × 10 is collected6It is a), 800rpm centrifugation 5min, are used
PBS is washed 3 times, and 70% ethanol for 2-8 DEG C is fixed, and after 4 DEG C are stayed overnight, then washs 2 times cells, 1000rpm centrifugations again with PBS
5min, abandoning supernatant, progress PI (propidium iodide) dyeing and FITC (Fluorescein isothiocynate) dye and TRITC (tetramethyls
Rhodamine isothiocyanate) dyeing, the analysis of flow cytometry analysis Cellular cycle and apoptosis, while blank control group is set, blank
Control group is the physiological saline of equivalent.The result is shown in table 1, Fig. 3, Fig. 4, Fig. 5, Fig. 6.
Cell cycle result after 1 Chinese medicine composition of table effect NB4
Table 1 is shown, the apoptotic peak of FRBS according to the numerical value of sub-G1 the results show that FRBS has inhibitory action to NB4 cells
It is worth apparently higher than blank group；After the FRBS and RIF of identical arsenic concentration (2 μ g/ml) handle NB4 cells 24h respectively, FRBS inductions
The apoptosis capacity of NB4 cells is better than the apoptosis-induced effects of RIF；In addition, the FRBS of 2 μ g/ml of arsenic concentration and 4 μ g/ml can induce
The apoptosis of NB4 cells, its apoptosis-induced ability are presented dose dependent, are proportionate with concentration；Found from the numerical value of G0/G1,
After FRBS and RIF groups processing 24h, the numerical value of G0/G1 reduces, gradually to G2 phase transition, and with the increase of arsenic concentration in FRBS,
The numerical value of G0/G1 is less and less；In the case of identical arsenic concentration, RIF is more than the retardation of NB4 g2 phases the resistance of FRBS
Stagnant effect, occurs in the G2 phases retardation of cell NB4 this further illustrates FRBS and RIF.
Fig. 3, Fig. 4, Fig. 5, Fig. 6 show that arsenic concentration is that growths of the FRBS of 2 μ g/ml and 4 μ g/ml to NB4 cells has suppression
Effect, and the apoptosis rate in early stage is respectively 39.1%, 72.9%, apoptosis rate increases with the increase of arsenic concentration；Identical arsenic is dense
The FRBS and RIF of degree are respectively 39.1% and 27.0% to the early apoptosis rate of NB4, illustrate apoptosis-induced energy of the FRBS to NB4
Power is better than apoptosis-induced abilities of the RIF to NB4, and apoptosis-induced ability is proportionate with arsenic concentration.
4 Chinese medicine composition of embodiment detects the apoptosis of HL-60 cells
Biomaterial, reagent are the same as embodiment two.
After HL-60 cells are handled 24h with FRBS, RIF, cell suspension (1 × 10 is collected6It is a), 800rpm centrifugation 5min,
Washed 3 times with PBS, 70% ethanol for 2-8 DEG C is fixed, 4 DEG C overnight after, then wash 2 times cells again with PBS, 1000rpm from
Heart 5min, abandoning supernatant, progress PI (propidium iodide) dyeing and FITC (Fluorescein isothiocynate) dye and TRITC (tetramethyls
Base rhodamine isothiocyanate) dyeing, the analysis of flow cytometry analysis Cellular cycle and apoptosis, while blank control group is set, it is empty
White control group is the physiological saline of equivalent.The result is shown in table 2 and Fig. 7, Fig. 8, Fig. 9, Figure 10.
Cell cycle result after 2 Chinese medicine composition of table effect HL-60
Table 2 is shown, the apoptosis of FRBS according to the numerical value of sub-G1 the results show that FRBS has inhibitory action to HL-60 cells
Peak value is apparently higher than blank group；After the FRBS and RIF of identical arsenic concentration (2 μ g/ml) handle NB4 cells 24h respectively, FRBS is lured
The apoptosis capacity for leading HL-60 cells is better than the apoptosis-induced abilities of RIF；In addition, the equal energy of FRBS of arsenic concentration 2 μ g/ml and 4 μ g/ml
The apoptosis of HL-60 cells cell, its apoptosis-induced ability are presented dose dependent, are proportionate with concentration；From the numerical value of G0/G1
It was found that after the reason HL-60 of FRBS and RIF components other places, the numerical value of G0/G1 reduces, gradually to G2 phase transition, and with arsenic in FRBS
The increase of concentration, the numerical value of G0/G1 are less and less.
Fig. 7, Fig. 8, Fig. 9, Figure 10 show that arsenic concentration is that growths of the FRBS of 2 μ g/ml and 4 μ g/ml to HL-60 cells has
Inhibitory action, and apoptosis rate difference 31.9%, 59.3% late, apoptosis rate increases with the increase of concentration；Identical arsenic is dense
The FRBS and RIF of degree are respectively 31.9% and 10.4% to the apoptosis rate in HL-60 late periods, illustrate that inductions of the FRBS to HL-60 is withered
The ability of dying is better than apoptosis-induced abilities of the RIF to HL-60, and apoptosis-induced ability is proportionate with arsenic concentration.
In conclusion Chinese medicine composition not only significantly reduces the dosage of realgar, reduce toxicity, it is to acute early young grain
The therapeutic effect of leukaemia is better than compound realgar natural indigo tablet.And detected by cell cycle, WB, it can go out tumour cell
Now obvious apoptotic body, mostly occurs NB4 Apoptosis in early stage, mostly occurs late, pass through to the apoptosis of HL-60 cells
Contrast commercially available Treated with Compound Huangdai Tablets effect to understand, when arsenic concentration is commercially available 1/2 in invention Chinese medicine composition, still can
The identical therapeutic effect enough reached.
Claims (7)
1. a kind of Chinese medicine composition for treating leukaemia, it is characterised in that the Chinese medicine composition of the treatment leukaemia includes down
It is 33.5 to state proportioning：40：12：The indigo naturalis of 10m/m/m/v, Radix Salviae Miltiorrhizae, radix pseudostellariae and realgar microorganism extracting liquid.
2. the Chinese medicine composition for the treatment of leukaemia according to claim 1, it is characterised in that the realgar microorganism leaches
Total arsenic concentration of liquid is 461.4 μ g/ml.
3. the Chinese medicine composition for the treatment of leukaemia according to claim 1 or 2, it is characterised in that the realgar microorganism
Leachate is prepared by following methods：Take Fe containing 1g/L2+The 0K culture medium 90ml of ferrous sulfate, add realgar powder 0.5g,
The Thiobacillus ferrooxidans of 10ml domestications is inoculated with, is leached 25-30 days in 30 DEG C, pH1.8,130rpm constant temperature oscillation Shaking culture,
Precipitation is abandoned in filtering, obtains filtrate, and it is 7.0 to take the NaOH solution containing 2mol/L EDTA-2Na to adjust filtrate pH value, micro- up to realgar
Biochemical lixivium.
4. the Chinese medicine composition for the treatment of leukaemia according to claim 3, it is characterised in that the ferrous oxide sulphur bar
Bacterium is the Thiobacillus ferrooxidans of arsenic domestication, and domestication flow is：Thiobacillus ferrooxidans is taken, with 200 μ g sodium arsenites/100ml
9K culture mediums, 400 μ g sodium arsenites/100ml 9K culture mediums, 600 μ g sodium arsenites/100ml 9K culture mediums, 800 μ g arsenous
The domestication of sour sodium/100ml 9K culture mediums, 1600 μ g sodium arsenites/100ml 9K culture mediums gradient, then take again 0.1g realgars/
100ml 9K culture mediums, 0.2g realgars/100ml 9K culture mediums, 0.3g realgars/100ml 9K culture mediums, 0.5g realgars/100ml
9K culture mediums, 1.0g realgars/100ml 9K culture mediums gradient domestication, gradually reduce the amount of ferrous sulfate in culture medium afterwards, when
When the amount of ferrous sulfate is 3K, domestication Thiobacillus ferrooxidans is obtained.
5. the Chinese medicine composition for the treatment of leukaemia according to claim 1 or 2, it is characterised in that the treatment leukaemia
Chinese medicine composition include following raw materials：Indigo naturalis 33.5g, Radix Salviae Miltiorrhizae 40g, radix pseudostellariae 12g, realgar microorganism extracting liquid 10ml, institute
The total arsenic concentration for stating realgar microorganism extracting liquid is 461.4 μ g/ml.
6. the Chinese medicine composition for the treatment of leukaemia according to claim 1 is preparing treatment acute promyelocytic leukemia
Application in medicine.
A kind of 7. preparation method of the Chinese medicine composition for the treatment of leukaemia as claimed in claim 1, it is characterised in that the system
Preparation Method is as follows：
(1) radix pseudostellariae, indigo naturalis are taken, is crushed, is uniformly mixed spare；
(2) Radix Salviae Miltiorrhizae is taken, distilled water is added, is evaporated under reduced pressure to the filtrate of the Radix Salviae Miltiorrhizae of concentration, adds realgar microorganism extracting liquid, is dried
It is dry to obtain medicinal extract；
(3) medicinal extract for obtaining step (2) adds the radix pseudostellariae indigo naturalis mixture that step (1) obtains, and dries, and crushes, mixes well, and makes
Grain, is drying to obtain the Chinese medicine composition for the treatment of leukaemia.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610948385.3A CN107982315A (en) | 2016-10-26 | 2016-10-26 | A kind of Chinese medicine composition for treating leukaemia and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610948385.3A CN107982315A (en) | 2016-10-26 | 2016-10-26 | A kind of Chinese medicine composition for treating leukaemia and preparation method thereof |
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| CN111012799A (en) * | 2018-10-09 | 2020-04-17 | 兰州大学 | Application of realgar microbial leaching as PML-RAR α protein inhibitor |
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