CN103877112A - Pharmaceutical composition containing ginsenoside Rh2 and ophiopogonin B - Google Patents
Pharmaceutical composition containing ginsenoside Rh2 and ophiopogonin B Download PDFInfo
- Publication number
- CN103877112A CN103877112A CN201410136969.1A CN201410136969A CN103877112A CN 103877112 A CN103877112 A CN 103877112A CN 201410136969 A CN201410136969 A CN 201410136969A CN 103877112 A CN103877112 A CN 103877112A
- Authority
- CN
- China
- Prior art keywords
- ginsenoside
- ophiopogonin
- add
- pharmaceutical composition
- ethyl acetate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to the technical field of medicines, and discloses a pharmaceutical composition containing ginsenoside Rh2 and ophiopogonin B. The pharmaceutical composition comprises the following active components: ginsenoside Rh2 and ophiopogonin B. The pharmaceutical composition containing the ginsenoside Rh2 and the ophiopogonin B provided by the invention has excellent stability. The pharmacological test shows that the pharmaceutical composition disclosed by the invention has good pharmacological action.
Description
Technical field
The invention belongs to medical technical field, be specifically related to pharmaceutical composition of a kind of ginsenoside Rh2 of containing, ophiopogonin B and preparation method thereof.
Background technology
The first-class rare Chinese medicine of Panax, main effect is: " promoting the production of body fluid to quench thirst, calms the nerves and increase intelligence for strongly invigorating primordial QI, invigorating the spleen to benefit the lung ".Numerous disease can not only be treated and prevent to Radix Ginseng, is also the good merchantable brand of health care.Can be divided into wild ginseng, Park Ginseng, Jilin ginseng, Korean ginseng according to the Radix Ginseng place of production and concocting method, Radix Ginseng, Radix Ginseng, Radix Ginseng Rubra etc.Radix Ginseng Rubra is that Radix Ginseng processes through steaming.
Nineteen eighty-three, Beichuan merit is isolated ginsenoside Rh2 first from Korea Radix Ginseng Rubra.By a large amount of research, now confirm that ginsenoside Rh2 can break up or apoptosis by induction, suppress propagation and the growth of kinds of tumor cells, and prove very low to normal cytotoxic effect; And ginsenoside Rh2 has the effect of good alleviating physical fatigue.
Summary of the invention
The object of this invention is to provide a kind of new pharmaceutical composition, said composition comprises ginsenoside Rh2 and ophiopogonin B, by extensive and deep research, inventor finds that the two carries out prescription at certain weight ratio and have collaborative pharmacological action, be that the two can prescription under certain part by weight, and have excellent pharmacological effect effect, at saving resource, have on the basis of better drug action, active ingredient composition has better safety.
The present invention is achieved through the following technical solutions.
A pharmaceutical composition that contains ginsenoside Rh2, ophiopogonin B, this pharmaceutical composition active component is: ginsenoside Rh2, ophiopogonin B.
Described pharmaceutical composition preferably ginseng saponin Rh210-15 weight portion, ophiopogonin B 5-10 weight portion.
Pharmaceutical composition described above is preferably: ginsenoside Rh2 10-15 weight portion, ophiopogonin B 5-10 weight portion, arginine 58-63 weight portion.
Described pharmaceutical composition is: ginsenoside Rh2 10-15 weight portion, ophiopogonin B 5-10 weight portion, arginine 59-61 weight portion.
Described ginsenoside Rh2's 13 weight portions, ophiopogonin B 8 weight portions, arginine 60 weight portions.
Pharmaceutical composition described above is prepared into tablet or ejection preparation.
The preparation method of ejection preparation described above is:
(1) get arginine, etc. the water of quality, stir, add ginsenoside Rh2, ophiopogonin B, be heated to 50～60 ℃, stirring and dissolving, with containing 9-11% salt aqueous acid adjust pH to 3.8～4.8, adds water to full dose;
(2) add the active carbon of dosing amount 0.1-0.3%g/ml, stir 25-35 minute, coarse filtration takes off charcoal;
(3) cross 0.22 μ m filter membrane, obtain fine straining liquid;
(4) embedding;
(5) 118-125 ℃ of pressure sterilizing 13-18 minute, check, packing, warehouse-in.
Described pharmaceutical composition, wherein the preparation method of tablet is:
(1) ginsenoside Rh2, ophiopogonin B are stirred after mixing, obtain pulverulent solids;
(2) powder of step (1) gained is mixed homogeneously with lactose, carboxymethyl starch sodium and micropowder silica gel, add arginine to make suitable soft material, 20 mesh sieves are granulated;
(3) the dry granule of carboxymethyl starch sodium and micropowder silica gel and step (2) gained mixes, and tabletting, obtains tablet.
Pharmaceutical composition described above, wherein ginsenoside Rh2's preparation method, comprising:
(1) under stirring condition, in ethyl acetate, first add kieselguhr, then add ginsenoside extract, in 78-85 ℃ of reflux 0.5-1.5 hour, filter, concentrated; Add diatomaceous amount be ginsenoside extract weight 0.8-1.2 doubly, the liquid-solid ratio of ethyl acetate and ginsenoside extract is 15-20ml/g;
(2) add distilled water remove impurity again, separate ethyl acetate layer, ethyl acetate layer peracidity alumina column, uses water saturation eluent ethyl acetate, collects eluent;
(3) eluent is evaporated to without ethyl acetate taste, add ethanol make it dissolve, then add water to make containing alcohol amount be 60%-70%, leave standstill, make to separate out coarse crystallization, filter, get coarse crystallization, dry;
(4) coarse crystallization is used ethanol and re-crystallizing in ethyl acetate successively, obtains finished product ginsenoside Rh2.
Preparation Example
Embodiment 1
The preparation of ginsenoside Rh2's extract:
Under stirring condition, in 750ml ethyl acetate, first add kieselguhr 40g, then add ginsenoside extract 50g, in 85 ℃ of reflux 0.5 hour, filter, concentrated, it is 5ml:lg that filtrate is concentrated into liquid-solid ratio; Toward the distilled water remove impurity that adds approximately 1/10 volume in concentrated solution, separate ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:4, aluminium oxide is 200 orders, and the weight ratio of applied sample amount and aluminium oxide is 1:7, uses water saturation eluent ethyl acetate, elution speed is l/4BV/h, collects eluent; Eluent is evaporated to without ethyl acetate taste, add appropriate ethanol make it dissolve, then add water to make containing alcohol amount be 60%, leave standstill, make to separate out coarse crystallization, filter, get coarse crystallization, dry; Coarse crystallization is used ethanol and re-crystallizing in ethyl acetate successively, obtains finished product ginsenoside Rh2 extract.Sample detection ginsenoside Rh2's content is 99.2%.
Embodiment 2
The preparation of ginsenoside Rh2's extract:
Under stirring condition, in 1000ml ethyl acetate, first add kieselguhr 40g, then add ginsenoside extract 50g, in 79 ℃ of reflux 1.5 hours, filter, concentrated, it is 4ml:lg that filtrate is concentrated into liquid-solid ratio; Toward the distilled water remove impurity that adds approximately 1/9 volume in concentrated solution, separate ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:4, aluminium oxide is 200 orders, and the weight ratio of applied sample amount and aluminium oxide is 1:7, uses water saturation eluent ethyl acetate, elution speed is l/4BV/h, collects eluent; Eluent is evaporated to without ethyl acetate taste, add appropriate ethanol make it dissolve, then add water to make containing alcohol amount be 60%, leave standstill, make to separate out coarse crystallization, filter, get coarse crystallization, dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, obtains finished product ginsenoside Rh2 extract.Sample detection ginsenoside Rh2's content is 99.6%.
Embodiment 3
The preparation of ginsenoside Rh2's extract:
Under stirring condition, in 850ml ethyl acetate, first add kieselguhr 60g, then add ginsenoside extract 50g, in 80 ℃ of reflux I hour, filter, concentrated, it is 3ml:lg that filtrate is concentrated into liquid-solid ratio; Toward the distilled water remove impurity that adds approximately 1/11 volume in concentrated solution, separate ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:10, aluminium oxide is 300 orders, and the weight ratio of applied sample amount and aluminium oxide is 1:10, uses water saturation eluent ethyl acetate, elution speed is l/2BV/h, collects eluent; Eluent is evaporated to without ethyl acetate taste, add appropriate ethanol make it dissolve, then add water to make containing alcohol amount be 70%, leave standstill, make to separate out coarse crystallization, filter, get coarse crystallization, dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, obtains finished product ginsenoside Rh2 extract.Sample detection ginsenoside Rh2's content is 99.7%.
Embodiment 4
The preparation of ginsenoside Rh2's extract:
Under stirring condition, in 900ml ethyl acetate, first add kieselguhr 45g, then add ginsenoside extract 50g, in 79 ℃ of reflux I hour, filter, concentrated, it is 3.5ml:lg that filtrate is concentrated into liquid-solid ratio; Toward the distilled water remove impurity that adds approximately 1/10 volume in concentrated solution, separate ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:7, aluminium oxide is 300 orders, and the weight ratio of applied sample amount and aluminium oxide is 1:8, uses water saturation eluent ethyl acetate, elution speed is l/2BV/h, collects eluent; Eluent is evaporated to without ethyl acetate taste, add appropriate ethanol make it dissolve, then add water to make containing alcohol amount be 70%, leave standstill, make to separate out coarse crystallization, filter, get coarse crystallization, dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, obtains finished product ginsenoside Rh2 extract.Sample detection ginsenoside Rh2's content is 99.6%.
Embodiment 5
The preparation of ginsenoside Rh2's extract:
Under stirring condition, in 900ml ethyl acetate, first add kieselguhr 55g, then add ginsenoside extract 50g, in 78 ℃ of reflux I hour, filter, concentrated, it is 3.5ml:lg that filtrate is concentrated into liquid-solid ratio; Toward the distilled water remove impurity that adds approximately 1/10 volume in concentrated solution, separate ethyl acetate layer, ethyl acetate layer peracidity alumina column, the blade diameter length ratio of acidic alumina column is 1:4, aluminium oxide is 300 orders, and the weight ratio of applied sample amount and aluminium oxide is 1:8, uses water saturation eluent ethyl acetate, elution speed is l/2BV/h, collects eluent; Eluent is evaporated to without ethyl acetate taste, add appropriate ethanol make it dissolve, then add water to make containing alcohol amount be 65%, leave standstill, make to separate out coarse crystallization, filter, get coarse crystallization, dry; Coarse crystallization is used ethyl alcohol recrystallization 3 times and re-crystallizing in ethyl acetate 1 time successively, obtains finished product ginsenoside Rh2 extract.Sample detection ginsenoside Rh2's content is 99.7%.
Embodiment 6
The preparation of ginsenoside Rh2, ophiopogonin B injection:
Take arginine 58g, etc. the water of quality, stir, the ginsenoside Rh2 10g, the ophiopogonin B 5g that add embodiment 1 to prepare, be heated to 50 ℃, stirring and dissolving, with containing 9% salt aqueous acid adjust pH to 4.6, adds water to full dose, stirs; Add the active carbon of dosing amount 0.l% (g/ml) again, stir 25 minutes, coarse filtration takes off charcoal; Cross 0.22 μ m filter membrane, obtain fine straining liquid; Embedding, every bottled amount 2mL; 118 ℃ of pressure sterilizings 18 minutes, check, packing, warehouse-in.
Embodiment 7
The preparation of ginsenoside Rh2, ophiopogonin B injection:
Take arginine 63g, etc. the water of quality, stir, the ginsenoside Rh2 12g, the ophiopogonin B 6g that add embodiment 2 to prepare, be heated to 55 ℃, stirring and dissolving, with containing 11% salt aqueous acid adjust pH to 4.2, the 70g that adds water, stirs; Add the active carbon of dosing amount 0.3% (g/ml) again, stir 35 minutes, coarse filtration takes off charcoal; Cross 0.22 μ m filter membrane, obtain fine straining liquid; Embedding, every bottled amount 2mL; 125 ℃ of pressure sterilizings 13 minutes, check, packing, warehouse-in.
Embodiment 8
The preparation of ginsenoside Rh2, ophiopogonin B injection:
Take arginine 59g, etc. the water of quality, stir, the ginsenoside Rh2 13g, the ophiopogonin B 8g that add embodiment 3 to prepare, be heated to 60 ℃, stirring and dissolving, with containing 10% salt aqueous acid adjust pH to 4.0, adds water to full dose, stirs; Add the active carbon of dosing amount 0.l% (g/ml) again, stir 30 minutes, coarse filtration takes off charcoal; Cross 0.22 μ m filter membrane, obtain fine straining liquid; Embedding, every bottled amount 2mL; 121 ℃ of pressure sterilizings 15 minutes, check, packing, warehouse-in.
Embodiment 9
The preparation of ginsenoside Rh2, ophiopogonin B injection:
Take arginine 61g, etc. the water of quality, stir, the ginsenoside Rh2 14g, the ophiopogonin B 8g that add embodiment 4 to prepare, be heated to 50 ℃, stirring and dissolving, with containing 10% salt aqueous acid adjust pH to 3.8, adds water to full dose, stirs; Add the active carbon of dosing amount 0.l% (g/ml) again, stir 30 minutes, coarse filtration takes off charcoal; Cross 0.22 μ m filter membrane, obtain fine straining liquid; Embedding, every bottled amount 2mL; 121 ℃ of pressure sterilizings 15 minutes, check, packing, warehouse-in.
Embodiment 10
The preparation of ginsenoside Rh2, ophiopogonin B injection:
Take arginine 60g, etc. the water of quality, stir, the ginsenoside Rh2 15g, the ophiopogonin B 10g that add embodiment 5 to prepare, be heated to 60 ℃, stirring and dissolving, with containing 10% salt aqueous acid adjust pH to 4.8, adds water to full dose, stirs; Add the active carbon of dosing amount 0.l% (g/ml) again, stir 30 minutes, coarse filtration takes off charcoal; Cross 0.22 μ m filter membrane, obtain fine straining liquid; Embedding, every bottled amount 2mL; 121 ℃ of pressure sterilizings 15 minutes, check, packing, warehouse-in.
Embodiment 11
The preparation of ginsenoside Rh2, ophiopogonin B tablet:
(1) by stirring after ginsenoside Rh2 10g, ophiopogonin B 5g mixing, obtain pulverulent solids;
(2) powder of step (1) gained is mixed homogeneously with lactose 55g, carboxymethyl starch sodium 5g and micropowder silica gel 3.6g, add arginine 58g to make suitable soft material, 20 mesh sieves are granulated;
(3) the dry granule of carboxymethyl starch sodium 4.8g and micropowder silica gel 2.3g and step (2) gained mixes, and tabletting, obtains tablet.
Embodiment 12
The preparation of ginsenoside Rh2, ophiopogonin B tablet:
(1) by stirring after ginsenoside Rh2 15g, ophiopogonin B 10g mixing, obtain pulverulent solids;
(2) powder of step (1) gained is mixed homogeneously with lactose 55g, carboxymethyl starch sodium 2.6g and micropowder silica gel 4.3g, add arginine 63g to make suitable soft material, 20 mesh sieves are granulated;
(3) the dry granule of carboxymethyl starch sodium 5.2g and micropowder silica gel 1.5g and step (2) gained mixes, and tabletting, obtains tablet.
Embodiment 13
The preparation of ginsenoside Rh2, ophiopogonin B tablet:
(1) by stirring after ginsenoside Rh2 12g, ophiopogonin B 8g mixing, obtain pulverulent solids;
(2) powder of step (1) gained is mixed homogeneously with lactose 55g, carboxymethyl starch sodium 10.8g and micropowder silica gel 8.7g, add arginine 60g to make suitable soft material, 20 mesh sieves are granulated;
(3) the dry granule of carboxymethyl starch sodium 8.3g and micropowder silica gel 2.8g and step (2) gained mixes, and tabletting, obtains tablet.
Embodiment 14
The preparation of ginsenoside Rh2, ophiopogonin B tablet:
(1) by stirring after ginsenoside Rh2 12g, ophiopogonin B 6g mixing, obtain pulverulent solids;
(2) powder of step (1) gained is mixed homogeneously with lactose 55g, carboxymethyl starch sodium 7.8g and micropowder silica gel 4.3g, add arginine 59g to make suitable soft material, 20 mesh sieves are granulated;
(3) the dry granule of carboxymethyl starch sodium 4.3g and micropowder silica gel 2.1g and step (2) gained mixes, and tabletting, obtains tablet.
Test example 1
This experimental example has been prepared ginsenoside Rh2 with reference to the method for following documents, and adopt method of testing (" content of Rh2 in Radix Ginseng by HPLC extract " (" Tianjin Science & Engineering Univ journal " the 19th in March, 2003 volume the 1st phase) of document, measure ginsenoside Rh2's content in ginsenoside extract, the results are shown in Table 1.
Ginsenoside Rh2's extract that sample 1 is prepared for the method for the embodiment 1 with reference to patent 200910228462.8;
Ginsenoside Rh2's extract that sample 2 is prepared for the method for the embodiment 1 with reference to patent 200810233707.1;
Ginsenoside Rh2's extract that sample 3 is prepared for the method for the embodiment 1 with reference to patent 201110120780.X;
Ginsenoside Rh2's extract that sample 4 is prepared for the method for the embodiment 1 with reference to patent 200410021835.1;
Table 1 ginsenoside Rh2 extract purity test result
| Group | Sample 1 | Sample 2 | Sample 3 | Sample 4 |
| Ginsenoside Rh2's purity | 97.8% | 98.7% | 87.4% | 89.1% |
The measurement result of table 1 shows, compared with the prior art of above-mentioned patent application, in ginsenoside Rh2's extract prepared by the present invention, ginsenoside Rh2's content is high, can be used as the crude drug of injection, has greatly improved patient's drug safety.
Test example 2
This test example has been tested the stability of ginsenoside Rh2 extract prepared by the present invention.
Sample 1 is embodiment 1 product, and sample 2 is embodiment 2 products; Sample 3 is embodiment 5 products;
The ginsenoside Rh2 that sample 4 is 97.8% for the HPLC purity that adopts the method for patent 200910228462.8 embodiment 1 to obtain;
The ginsenoside Rh2 that sample 5 is 98.7% for the HPLC purity that adopts the method for patent 200810233707.1 embodiment 1 to obtain;
Sample is respectively got 1g, and this experiment is carried out according to 2010 editions second appendix XIXC medicine stability test guideline of Chinese Pharmacopoeia, and result is as follows:
Table 2 accelerated test result
| Sample | 0 month | 1 month | 2 months | 3 months | 6 months | 9 months |
?
| 1 | 99.0% | 99.1% | 98.9% | 98.8% | 98.6% | 98.5% |
| 2 | 99.6% | 99.6% | 99.5% | 99.3% | 99.2% | 99.0% |
| 3 | 99.7% | 99.6% | 99.6% | 99.5% | 99.3% | 99.2% |
| 4 | 97.8% | 93.8% | 91.7% | 90.5% | 90.1% | 89.9% |
| 5 | 98.7% | 94.6% | 92.6% | 91.2% | 90.9% | 90.1% |
Table 3 long-term test results
| Sample 1 | 0 month 99.0% | 3 months 98.8% | 6 months 98.6% | 9 months 98.5% | 12 months 98.2% | 18 months 98.1% |
| 2 | 99.6% | 99.3% | 99.2% | 99.0% | 98.9% | 98.6% |
| 3 | 99.7% | 99.5% | 99.3% | 99.2% | 99.0% | 98.9% |
| 4 | 97.8% | 97.5% | 94.1% | 92.9% | 91.5% | 90.4% |
| 5 | 98.7% | 97.2% | 95.9% | 93.7% | 92.4% | 90.5% |
Ginsenoside Rh2's provided by the invention content is higher, investigates by accelerated test and experiment for long-term stability, and result shows that ginsenoside Rh2's extract of the present invention has better stability with respect to prior art, accelerates, long term test purity changes of contents is little.
Test example 3 compositions screening test-anti-fatigue tests
Trial drug:
Test 1 group: ginsenoside Rb1 13mg, ophiopogonin B 8mg.
Test 2 groups: ginsenoside Rb1 13mg, ophiopogonin D 8mg.
Test 3 groups: ginsenoside Rd 13mg, ophiopogonin B 8mg.
Test 4 groups: ginsenoside Rd 13mg, ophiopogonin D 8mg.
Test 5 groups: ginsenoside Rg1 13mg, ophiopogonin B 8mg.
Test 6 groups: ginsenoside Rg1 13mg, ophiopogonin D 8mg.
Test 7 groups: ginsenoside Rh2 13mg, ophiopogonin B 8mg.
Test 8 groups: ginsenoside Rh2 13mg, ophiopogonin D 8mg.
Experimental animal: animal 2-3 monthly age, Kunming kind male and healthy mice, body weight (25 ± 3) g.
Test method: laboratory animal mice is divided into group, i.e. different pharmaceutical group and matched group, every group of 12 mices, adopt administration by gavage, and every day, matched group gave equivalent distilled water with 0.35g/kg dosage to mouse stomach different pharmaceutical.1 time/d, 21d continuously.Within the 22nd day, carry out the mensuration of every resisting fatigue index.
Motion model matched group and experimental group start to carry out adaptability swimming instruction on the 15th day in gavage, swimming time is started by 10min/d, increase progressively 5min to the every day 20 days, power exhausts criterion for the 8s that sinks under water by Mouse Weight 5% swimming with a load attached to the body to mice does not float, and after pulling out, cannot complete righting reflex.Swimming time: last is subject to after reagent 30min to mice, adopting load mode is that a quality is the lead of Mouse Weight 5% in Mus tail, mice is placed in to the swimming trunk of depth of water 30cm, the temperature of water is controlled at 30 ℃ of left and right.Forced swimming to power exhausts (till the 8s that sinks under water is motionless), records swimming time.The mensuration of serum urea nitrogen: the 1. preparation of serum: last is subject to after reagent 30min to mice, mice is not born a heavy burden and is placed in the swimming trunk of depth of water 30cm, the temperature of water is controlled at 30 ℃ of left and right, swimming 90min, afterbody blood sampling at once after rest 30min, about 0.5ml that takes a blood sample (not adding anticoagulant).Put 4 ℃ of about 3h of refrigerator, the centrifugal 15min of 2500r/min after hemopexis, gets serum and measures serum urea nitrogen value.2. determination of urea nitrogen method: press document [comparison [J] of Wei Xu east .3 kind serum urea nitrogen determination method. Chongqing Medical .1997,26 (4): 246.] method measures.The mensuration of hepatic glycogen: the 1. extraction of hepatic glycogen: last is subject to after reagent 30min to mice, de-neck is put to death at once, gets liver, sucks the blood adhering to rapidly with filter paper.Take about 1g, put in mortar, add and clean fine sand a little and 10% trichloroacetic acid 1ml and grind.Add again 5% trichloroacetic acid 2ml and continue to grind, till fully having worn into meat paste shape to liver organization.Then with the centrifugal 10min of 2500r/min.Supernatant proceeded to another centrifuge tube and measures volume, add 95% ethanol of same volume, after mixing, leaving standstill 10min.Now glycogen becomes flocculent deposit to separate out.Precipitation solution is with the centrifugal 10min of 2500r/min.Abandoning supernatant, and centrifuge tube is inverted in to 2min on filter paper.In precipitation, add distilled water 1ml Glass rod to stir and be precipitated to dissolving.2. the mensuration of hepatic glycogen: press literature method anthrone-sulphate method and measure hepatic glycogen content, every group is repeated 3 times, average.
Table 4 swimming time
| Group | Average time (min) |
| Matched group | 6.1±1.1 |
| 1 group of medicine | 9.9±1.2* |
| 2 groups of medicines | 8.8±2.0* |
| 3 groups of medicines | 8.9±2.1* |
| 4 groups of medicines | 8.4±1.7* |
| 5 groups of medicines | 8.8±1.2* |
| 6 groups of medicines | 9.7±1.1* |
| 7 groups of medicines | 13.3±2.2** |
| 8 groups of medicines | 9.9±1.7* |
Note: with relatively * P<0.05 of matched group, * * P<0.01.
Table 5 serum urea nitrogen content
| Group | Serum urea nitrogen (C/mmolL-1) |
| Matched group | 14.3±0.6 |
| 1 group of medicine | 10.5±0.2* |
| 2 groups of medicines | 10.9±0.5* |
| 3 groups of medicines | 10.2±0.9* |
| 4 groups of medicines | 9.0±0.3* |
| 5 groups of medicines | 10.2±1.1* |
| 6 groups of medicines | 9.8±0.7* |
| 7 groups of medicines | 7.0±0.3** |
| 8 groups of medicines | 10.4±0.6* |
Note: with relatively * P<0.05 of matched group, * * P<0.01.
Conclusion (of pressure testing): above-mentioned preliminary experiment shows, ginsenoside Rh2 and ophiopogonin B combination have good resisting oxygen lack.
The research of test example 4 to treatment of viral myocarditis in mice effect
1, laboratory animal: GBI/J mice, 18 ± 2g/, male and female half and half.
2, for reagent thing:
Test 1 group: ginsenoside Rh2 10mg, ophiopogonin B 8mg.
Test 2 groups: ginsenoside Rh2 15mg, ophiopogonin B 8mg.
Test 3 groups: ginsenoside Rh2 21mg, ophiopogonin B 8mg.
3, experimental technique:
3.1 observation index: electrocardiogram, serum cardiac antibody fluorescent detects, cardiac muscular tissue's sections observation.
3.2 experimental techniques: will buy mice adaptability back and raise after 1 week, and be divided at random 4 groups, 10 every group, male and female half and half, are divided into: matched group, test 1-3 group.
Myocarditis Model copies and confirms: after getting 60 of 6 groups of mices with Coxsackie B virus suspension to mice collunarium, every each 0.3ml, every day 1 time, continuous 12 days, get at random 2 mices for every group, first make electrocardiographic criteria limb lead and detect, prompting blank group normal ECG, copy 6 groups of Myocarditis Model, electrocardiogram is all undesired, shows as decreased heart rate, and average heart rate is 500 ± 40 beats/min, sinus rhythm, irregularity of pulse, the room that takes place frequently is early rich, is premature ventricular beat, QS ripple, ST section moves down 0.1mv; Get blood as eye socket respectively again, centrifugalize serum, the conventional method film-making detecting by fluorescent is made heart antibody fluorescent and is detected under luminescence microscope, and blank group shows negative, and the group that copies Myocarditis Model all shows the positive.Dissect taking-up heart and do cardiac muscular tissue's section; The visible extensive swelling of myocardial cell, interstitial edema and mononuclear cell, lymphocytic infiltration under high power lens.Confirm that 4 group murine viral myocarditis models copy successfully.
Experimental therapy method: 4 groups of mices are made to treatment controlled trial by designing requirement, and matched group gives normal saline, preparation for treating group dosage 0.90mg/kg/day of the present invention, tail intravenously administrable, every day 1 time, treats 12 days continuously.
4, experimental therapy result:
4.1 Electrocardiographies: 4 groups of mices are first made to electrocardiogram lead and detect, result of the test: the results are shown in Table 6.
The comparison of the each treated animal ECG change of table 6
Note: with relatively * * P<0.01 of blank group, * P<0.05.
4.2 heart antibody fluorescents detect: 4 groups of mices are got to blood as eye socket respectively again, and separation of serum, the multiplex luminescence microscope of conventional method film-making detecting by fluorescent detects, and the results are shown in Table 7.
The comparison of table 7 heart antibody fluoroscopic examination
| Group | Animal number of elements | Positive | Negative |
| Blank group | 10 | 10 | 0 |
| Test 1 group | 10 | 0 | 10 |
| Test 2 groups | 10 | 0 | 10 |
| Test 3 groups | 10 | 3 | 7 |
Conclusion: preparation for treating group of the present invention and matched group comparison, the heart antibody variation of turning out cloudy, has utmost point significant difference; Work as ginsenoside Rh2: ophiopogonin B not within the scope of 10-15:5-10 with matched group comparison, there were significant differences (P<0.05), therefore applicant selects ginsenoside Rh2: ophiopogonin B weight ratio is 10-15:5-10.
Test example 6 adjuvant screening tests
Trial drug group:
Test 1 group: ginsenoside Rh2 13g, ophiopogonin B 8g, lysine 60g.
Test 2 groups: ginsenoside Rh2 13g, ophiopogonin B 8g, histidine 60g.
Test 3 groups: ginsenoside Rh2 13g, ophiopogonin B 8g, PEG400 60g.
Test 4 groups: ginsenoside Rh2 13g, ophiopogonin B 8g, arginine 60g.
Test 5 groups: ginsenoside Rh2 13g, ophiopogonin B 8g, Liquid Macrogol 60g.
Preparation method: claim adjuvant, etc. the water of quality, stir, add ginsenoside Rh2 13g, ophiopogonin B 8g, be heated to 60 ℃, stirring and dissolving, with containing 10% salt aqueous acid adjust pH to 4.0, adds water to full dose, stirs; Add the active carbon of dosing amount 0.l% (g/ml) again, stir 30 minutes, coarse filtration takes off charcoal; Cross 0.22 μ m filter membrane, obtain fine straining liquid; Embedding, every bottled amount 2mL; 121 ℃ of pressure sterilizings 15 minutes, check, packing.
Detection method: ginsenoside Rh2's reference substance 3mg, ophiopogonin B reference substance 2mg are got in the preparation of reference substance solution, accurately weighed, add methanol to scale, shakes up after fully dissolving, and to obtain final product.
The preparation of standard curve
Precision measures ginsenoside Rh2's reference substance solution 10 μ l, 20 μ l, 40 μ l, 60 μ l, 80 μ l, 100 μ l, with ophiopogonin B reference substance solution 10 μ l, 20 μ l, 40 μ l, 60 μ l, 80 μ l, 100 μ l, put respectively in 10ml tool plug test tube, wave most solvent, precision adds 5% vanillin glacial acetic acid solution-perchloric acid (2:8) mixed solution 1ml of (facing with now joining), put in 60 ℃ of water-baths and heat 15 minutes, take out, put in ice bath cooling, precision adds glacial acetic acid 5ml, shake up, immediately according to ultraviolet visible spectrophotometry (appendix V A of Chinese Pharmacopoeia version in 2010), at 544nm, 483nm wavelength place measures absorbance, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.
Algoscopy precision measures this product 1ml, is added on (D101 post 1.5cm × 12cm) in pretreated macroporous resin column, and first water 25ml eluting, discards water liquid.Use again 75% ethanol 60ml eluting, collect eluent, evaporate to dryness, residue adds dissolve with ethanol and is transferred in 10ml volumetric flask, adds ethanol dilution to scale, shakes up, as need testing solution; Precision measures 1ml and puts in tool plug test tube, evaporate to dryness, the method under sighting target directrix curve preparation, from " precision adds the mixed solution 1ml of 5% vanillin glacial acetic acid solution-perchloric acid (2:8) ", measure absorbance, the concentration of reading need testing solution from standard curve in accordance with the law, calculate, to obtain final product.
Result of the test: in table 8.
Result is investigated in the test of table 8 prescription screening
| Prescription | Ginsenoside Rh2's content % | Ophiopogonin B content % |
| 1 | 88.3 | 86.4 |
| 2 | 89.5 | 90.6 |
| 3 | 91.0 | 95.7 |
| 4 | 99.6 | 99.8 |
| 5 | 90.6 | 87.9 |
Conclusion (of pressure testing): the preparation after stability test, when arginine is less as ginsenoside Rh2, the ophiopogonin B changes of contents of the preparation of adjuvant, conform to quality requirements, therefore, preferably arginine is as adjuvant.
Claims (8)
1. contain a pharmaceutical composition for ginsenoside Rh2, ophiopogonin B, it is characterized in that pharmaceutical composition active component is: ginsenoside Rh2, ophiopogonin B.
2. pharmaceutical composition according to claim 1, wherein ginsenoside Rh2 10-15 weight portion, ophiopogonin B 5-10 weight portion.
3. according to pharmaceutical composition described in claim 1 or 2, wherein pharmaceutical composition is: ginsenoside Rh2 10-15 weight portion, ophiopogonin B 5-10 weight portion, arginine 58-63 weight portion.
4. compositions according to claim 1, described pharmaceutical composition is: ginsenoside Rh2 10-15 weight portion, ophiopogonin B 5-10 weight portion, arginine 59-61 weight portion.
5. compositions according to claim 1, is characterized in that, described ginsenoside Rh2's 13 weight portions, ophiopogonin B 8 weight portions, arginine 60 weight portions.
6. according to a kind of pharmaceutical composition described in claim 1-5 any one, it is characterized in that pharmaceutical composition is prepared into pharmaceutical preparation, pharmaceutical preparation comprises injection.
7. a kind of pharmaceutical composition according to claim 6, wherein the preparation method of injection is:
(1) get arginine, etc. the water of quality, stir, add ginsenoside Rh2, ophiopogonin B, be heated to 50-60 ℃, stirring and dissolving, with containing 9-11% salt aqueous acid adjust pH to 3.8-4.8, add water;
(2) add the active carbon of dosing amount 0.1-0.3%g/ml, stir 25-35 minute, coarse filtration takes off charcoal;
(3) cross 0.22 μ m filter membrane, obtain fine straining liquid;
(4) embedding;
(5) 118-125 ℃ of pressure sterilizing 13-18 minute, check, packing, warehouse-in.
8. according to the pharmaceutical composition described in claim 1-5 any one, wherein ginsenoside Rh2's preparation method: first add kieselguhr in ethyl acetate under stirring condition, then add ginsenoside extract, in 78-85 ℃ of reflux 0.5-1.5 hour, filter, concentrated; Add diatomaceous amount be ginsenoside extract weight 0.8-1.2 doubly, the liquid-solid ratio of ethyl acetate and ginsenoside extract is 15-20ml/g; Add distilled water remove impurity again, separate ethyl acetate layer, ethyl acetate layer peracidity alumina column, uses water saturation eluent ethyl acetate, collects eluent; Eluent is evaporated to without ethyl acetate taste, add ethanol make it dissolve, then add water to make containing alcohol amount be 60-70%, leave standstill, make to separate out coarse crystallization, filter, get coarse crystallization, dry; Coarse crystallization is used ethanol and re-crystallizing in ethyl acetate successively, both.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410136969.1A CN103877112B (en) | 2014-04-06 | 2014-04-06 | A kind of pharmaceutical composition containing ginsenoside Rh2, ophiopogonin B |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410136969.1A CN103877112B (en) | 2014-04-06 | 2014-04-06 | A kind of pharmaceutical composition containing ginsenoside Rh2, ophiopogonin B |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103877112A true CN103877112A (en) | 2014-06-25 |
| CN103877112B CN103877112B (en) | 2016-03-16 |
Family
ID=50946449
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410136969.1A Expired - Fee Related CN103877112B (en) | 2014-04-06 | 2014-04-06 | A kind of pharmaceutical composition containing ginsenoside Rh2, ophiopogonin B |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103877112B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110693898A (en) * | 2019-11-23 | 2020-01-17 | 吉林省蔚来生物科技有限公司 | A pharmaceutical composition containing ginsenoside |
| CN115531401A (en) * | 2022-10-12 | 2022-12-30 | 成都普睿法药物研发有限公司 | Method for improving stability of rare ginsenoside |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1682926A (en) * | 2005-03-04 | 2005-10-19 | 吴才梅 | Ginseng-ophiopogon root freeze-dried powder injection and its preparing method |
| CN1895540A (en) * | 2005-07-13 | 2007-01-17 | 成都迪康药物研究所 | Medicinal composition for treating cardiovascular disease, its making method and use |
| CN102988399A (en) * | 2012-12-21 | 2013-03-27 | 周志欢 | Medicine composition and application and preparation thereof |
-
2014
- 2014-04-06 CN CN201410136969.1A patent/CN103877112B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1682926A (en) * | 2005-03-04 | 2005-10-19 | 吴才梅 | Ginseng-ophiopogon root freeze-dried powder injection and its preparing method |
| CN1895540A (en) * | 2005-07-13 | 2007-01-17 | 成都迪康药物研究所 | Medicinal composition for treating cardiovascular disease, its making method and use |
| CN102988399A (en) * | 2012-12-21 | 2013-03-27 | 周志欢 | Medicine composition and application and preparation thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110693898A (en) * | 2019-11-23 | 2020-01-17 | 吉林省蔚来生物科技有限公司 | A pharmaceutical composition containing ginsenoside |
| CN115531401A (en) * | 2022-10-12 | 2022-12-30 | 成都普睿法药物研发有限公司 | Method for improving stability of rare ginsenoside |
| CN115531401B (en) * | 2022-10-12 | 2023-08-29 | 成都普睿法药物研发有限公司 | Method for improving stability of rare ginsenoside |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103877112B (en) | 2016-03-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10624938B2 (en) | Total flavone extract of flower of abelmoschus manihot L. medic and preparation method thereof | |
| CN103768534B (en) | A kind of Chinese medicine composition with antitumor activity | |
| CN103880910B (en) | A kind of preparation method and its usage of Cyclosiversigenin | |
| CN102885858B (en) | Method for preparing American cockroach extract for treating hepatitis B | |
| CN100545164C (en) | The preparation technology of disodium cantharidinate | |
| CN101033245B (en) | Preparation method and application of pedunculoside | |
| CN103877112B (en) | A kind of pharmaceutical composition containing ginsenoside Rh2, ophiopogonin B | |
| CN101099753A (en) | Preparation method and application for general saponin of cortex ilecis rotundae | |
| CN103830374B (en) | The application in hyperuricemia clearly of three leaf glycolipids | |
| CN117510443A (en) | Lemongrass extract L01, pharmaceutical composition and application thereof | |
| CN107281386B (en) | Method for preparing medicine for treating hypertension by using dendrobium officinale and radish seeds and application | |
| CN107693732B (en) | Medicine composition for treating hypertensive nephropathy and preparation method thereof | |
| CN101623437A (en) | Pulse activating preparation for injection and its preparation method | |
| CN102688248A (en) | Use of bufadienolide compound in preparing medicines for treating oral mucosal malignant tumors | |
| CN106581077A (en) | Preparing method of medicine for treating hepatitis B virus carrier | |
| CN101633661B (en) | Process for preparing sodium cantharidinate | |
| CN101032534A (en) | Method of preparing jiubiying total saponins and the application thereof | |
| CN107126450B (en) | A Chinese medicinal preparation containing extract and effective substance, and its preparation method and application | |
| CN108743676B (en) | Application method of white peony root in preparation of medicine for treating hypertension | |
| CN115364179B (en) | Traditional Chinese medicine composition for reducing blood uric acid as well as preparation method and application thereof | |
| CN110215469B (en) | Pharmaceutical composition for treating bile duct cancer | |
| CN113633712B (en) | Traditional Chinese medicine composition for treating gout and oral preparation based on traditional Chinese medicine composition | |
| CN101502536A (en) | Cedar total flavone as well as preparation method and medical use | |
| CN105596429B (en) | Preparation and application of lotus plumule total flavonoids for preventing and treating type II diabetes | |
| CN106265773B (en) | Lucidum spore powder glycopeptide extract and preparation method thereof and purposes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160316 Termination date: 20210406 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |