CN105044300A - Method for adjusting dosage of medicinal material coptidis rhizoma on basis of anti-dysentery bacillus effect equivalency factor - Google Patents
Method for adjusting dosage of medicinal material coptidis rhizoma on basis of anti-dysentery bacillus effect equivalency factor Download PDFInfo
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- coptidis rhizoma
- rhizoma coptidis
- coptis
- shigella dysenteriae
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- 230000000694 effects Effects 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 21
- 239000000463 material Substances 0.000 title abstract description 17
- 208000001848 dysentery Diseases 0.000 title abstract description 7
- 241000193830 Bacillus <bacterium> Species 0.000 title abstract 6
- 235000002991 Coptis groenlandica Nutrition 0.000 claims description 42
- 241000218202 Coptis Species 0.000 claims description 40
- 229940007046 shigella dysenteriae Drugs 0.000 claims description 38
- WITLAWYGGVAFLU-UHFFFAOYSA-N 3-(6-methoxy-1,3-benzodioxol-5-yl)-8,8-dimethylpyrano[2,3-f]chromen-4-one Chemical compound C1=CC(C)(C)OC2=CC=C(C(C(C3=CC=4OCOC=4C=C3OC)=CO3)=O)C3=C21 WITLAWYGGVAFLU-UHFFFAOYSA-N 0.000 claims description 18
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- XYHOBCMEDLZUMP-UHFFFAOYSA-N coptisine Chemical compound C1=C2C=C(C3=C(C=C4OCOC4=C3)CC3)[N+]3=CC2=C2OCOC2=C1 XYHOBCMEDLZUMP-UHFFFAOYSA-N 0.000 claims description 11
- PTPHDVKWAYIFRX-UHFFFAOYSA-N Palmatine Natural products C1C2=C(OC)C(OC)=CC=C2C=C2N1CCC1=C2C=C(OC)C(OC)=C1 PTPHDVKWAYIFRX-UHFFFAOYSA-N 0.000 claims description 10
- FPJQGFLUORYYPE-UHFFFAOYSA-N epiberberine Chemical compound C1=C2C=C(C3=C(C=C(C(=C3)OC)OC)CC3)[N+]3=CC2=C2OCOC2=C1 FPJQGFLUORYYPE-UHFFFAOYSA-N 0.000 claims description 10
- QUCQEUCGKKTEBI-UHFFFAOYSA-N palmatine Chemical compound COC1=CC=C2C=C(C3=C(C=C(C(=C3)OC)OC)CC3)[N+]3=CC2=C1OC QUCQEUCGKKTEBI-UHFFFAOYSA-N 0.000 claims description 10
- MXTLAHSTUOXGQF-UHFFFAOYSA-O Jatrorrhizine Chemical compound COC1=CC=C2C=C3C(C=C(C(=C4)O)OC)=C4CC[N+]3=CC2=C1OC MXTLAHSTUOXGQF-UHFFFAOYSA-O 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 4
- 241000037740 Coptis chinensis Species 0.000 claims description 2
- 244000247747 Coptis groenlandica Species 0.000 claims description 2
- 238000004128 high performance liquid chromatography Methods 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 9
- 229940079593 drug Drugs 0.000 abstract description 6
- 230000010354 integration Effects 0.000 abstract description 2
- 230000000144 pharmacologic effect Effects 0.000 abstract description 2
- 238000013441 quality evaluation Methods 0.000 abstract description 2
- 239000004480 active ingredient Substances 0.000 abstract 1
- 238000013139 quantization Methods 0.000 abstract 1
- 241000607764 Shigella dysenteriae Species 0.000 description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 239000000203 mixture Substances 0.000 description 8
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- 239000000243 solution Substances 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- YBHILYKTIRIUTE-UHFFFAOYSA-N berberine Chemical compound C1=C2CC[N+]3=CC4=C(OC)C(OC)=CC=C4C=C3C2=CC2=C1OCO2 YBHILYKTIRIUTE-UHFFFAOYSA-N 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 239000013558 reference substance Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
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- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical compound Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 description 3
- 241001411320 Eriogonum inflatum Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 229940093265 berberine Drugs 0.000 description 2
- QISXPYZVZJBNDM-UHFFFAOYSA-N berberine Natural products COc1ccc2C=C3N(Cc2c1OC)C=Cc4cc5OCOc5cc34 QISXPYZVZJBNDM-UHFFFAOYSA-N 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
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- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 description 1
- 238000012356 Product development Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
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- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
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- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a method for adjusting dosage of a medicinal material coptidis rhizoma on the basis of an anti-dysentery bacillus effect equivalency factor. The method comprises the following steps: (1), measuring the contents of active ingredients in the medicinal material coptidis rhizoma; (2), calculating an anti-dysentery bacillus effect equivalency factor of the coptidis rhizoma; (3), adjusting the dosage of the medicinal material coptidis rhizoma according to the anti-dysentery bacillus effect equivalency factor. According to the invention, a clinical function is used as guidance, the anti-dysentery bacillus effect equivalency factor can represent the coptidis rhizoma anti-dysentery bacillus pharmacological activity, and guide establishment and improvement of a clinical individualized medication standard; the problem of contribution difference of multiple component indexes to the efficacy of the coptidis rhizoma is solved, and comprehensiveness, quantization and integration of the coptidis rhizoma quality evaluation are realized; the efficacy of the coptidis rhizoma can be represented by detecting the contents of the coptidis rhizoma components directly, implementation is simple, easy and repeatable and the operability is high.
Description
Technical field
The present invention relates to a kind of method regulating Rhizoma Coptidis consumption based on anti-shigella dysenteriae effect equivalence factor.
Background technology
The coptis (CoptidisRhizoma) is ranunculaceae plant coptis CoptischinensisFranch., the dry rhizome of triangle leaf coptis CoptisdeltoideaC.Y.ChengetHsiao and Yun Lian CoptisteetaWall..Be usually used in heat-clearing and damp-drying drug, purging intense heat and detonicating, the composition relevant to its drug activity is biological alkali components, comprises jamaicin, epiberberine, coptisine, palmatine, jateorrhizine etc.Modern pharmacological research shows, the coptis has the effects such as antibacterial, antitumor, hypoglycemic, antiviral, anti-inflammatory, and wherein the inhibiting effect of the coptis to shigella dysenteriae effect traditional with it " only dysentery " has important correlativity.
As clinical bulk medicinal materials, how effectively to comment control to its clinical efficacy of guarantee and will promote that its product development is significant to coptis quality.Currently mainly realize commenting control to coptis quality with multiple index components quantitative test, but this method does not reflect the contribution degree difference of heterogeneity to the overall drug effect of medicinal material, the drug effect size ignored associated by heterogeneity is distinguished to some extent, is difficult to the quality situation evaluating different medicinal material during practical application.Concerning the coptis, act.std is carried out to it index components that quality checks on and is comprised jamaicin, coptisine, epiberberine, palmatine four alkaloids compositions, find in reality detects, in two batches of coptiss, a collection of content of berberine 8.2%, coptisine is 2.7%, and the content of berberine of another batch is 9.5%, do you when coptisine is 1.8%, now how judge the mass discrepancy of these two batches of medicinal materials by observing component content situation?
In addition, be different from chemicals, its constituent structure is clear, and structure-activity relationship is clear and definite, differentiates, checks, assay can directly as the index of therapeutic evaluation.And for Chinese medicine, its material intension is complicated, only detects component content and will cause a lot of limitation, need quality control index conscientiously to associate drug activity.If but only add these drug activity indexs at current component containing simple on the basis surveyed, then can increase difficulty when quality control standard is implemented from now on and complexity.Because increasing of each pharmacodynamics index, often kind of medicinal material can be caused to have more standards that will meet, more how loaded down with trivial details operation, and expend more experiment materials and animal used as test, this will certainly cause the waste of energy resources.Moreover the contribution degree difference problem that this multi objective Quality Control pattern is brought still does not solve, cannot illustrate wherein which index has prior association to quality of medicinal material.
Therefore, if a kind of directly deeply concerned " controlled effectively ", can not increase again Quality Control pattern and implementing difficulty, the coptis quality of clinical application can be instructed to comment control method comprehensively can be set up simultaneously, by significant to improving of the coptis and other Chinese medicine quality control standards.
Summary of the invention
The object of this invention is to provide a kind of method that anti-shigella dysenteriae effect equivalence factor (Effectequivalencyfactor, EEF) based on Rhizoma Coptidis regulates Rhizoma Coptidis consumption.
Anti-shigella dysenteriae effect equivalence factor based on Rhizoma Coptidis of the present invention regulates the method for Rhizoma Coptidis consumption to comprise the following steps:
(1) measure the content of the jamaicin (BER) in described Rhizoma Coptidis, epiberberine (EPI), coptisine (COP), palmatine (PAL) and jateorrhizine (JAT), namely measure X respectively
bER, X
ePI, X
cOP, X
pAL, X
jAT;
(2) utilize formula I to calculate the anti-shigella dysenteriae effect equivalence factor of Rhizoma Coptidis to be measured:
Formula I:EEF=3.352*X
bER+ 1.293*X
ePI+ 6.635*X
cOP+ 0.140*X
pAL+ 0.419*X
jAT-0.111;
Described EEF is the anti-shigella dysenteriae effect equivalence factor of Rhizoma Coptidis to be measured;
(3) formula II is utilized to calculate the actual amount of Rhizoma Coptidis:
Formula II is D
survey=D
target/ EEF; Wherein said D
surveyfor the actual amount of Rhizoma Coptidis to be measured, D
targetfor the target amount of Rhizoma Coptidis.
Of the present invention based on anti-shigella dysenteriae effect equivalence factor regulate the method for coptis consumption can not only by means of only measurement component content reflect its " effect ", the difficulty of quality standard when implementing can not be increased, have precisely can quantize, integratedly to reappear, comprehensively can operate, effective feature applicatory, and medicinal material clinical efficacy can be associated, according to anti-shigella dysenteriae effect equivalence factor, coptis consumption is debugged, ensure effect equivalent consistance, comment the research of the Innovation Integration of control standard and clinical application to instruct significant to the quality of Rhizoma Coptidis.
Embodiment
Summary of the invention is further illustrated below in conjunction with instantiation.
Method based on anti-shigella dysenteriae effect equivalence factor adjustment coptis consumption of the present invention can carry out activity rating to coptis sample and quantity is adjusted.
Instrument used in test: comminutor; Analytical balance (JM-B type, the inscription of Yuyao City discipline weigh calibration equipment company limited); Rotary Evaporators (R205B, Shensheng Science & Tech. Co., Ltd., Shanghai); Electric drying oven with forced convection (DHG9070A, Shanghai Yiheng Scientific Instruments Co., Ltd); The multiplex vacuum pump of recirculated water (SHB-B95, Zhengzhou Greatwall Scientific Industrial & Trading Co., Ltd.); Agilent1100 high performance liquid chromatograph (DAD detecting device), Chemstations chromatographic work station (Agilent company limited of the U.S.); BS210S type electronic balance (Beijing Sartorious company limited); Water is distilled water.
(1) content of the jamaicin in the Rhizoma Coptidis collected by measurement, epiberberine, coptisine, palmatine, jateorrhizine:
Rhizoma Coptidis collected by table 1
I () first prepares coptis water extract: get Rhizoma Coptidis removing impurity, be ground into meal after clean dry.Take each 50g of Rhizoma Coptidis powder of different batches, soak 30 minutes, decocting extracts 3 times, and (amount of water is respectively 10,10,8 times of medicinal material weight, each extraction 1.5,1.0,0.5 hours), filter while hot, merging filtrate, reduced pressure concentration, medicinal extract is gone in baking oven, 75 DEG C of oven dry, obtain coptis water extraction sample dry powder.
(ii) reference substance solution preparation: precision takes jamaicin, epiberberine, coptisine, palmatine, jateorrhizine are in right amount each, is placed in brown measuring bottle respectively, adds hydrochloric acid-methanol (1:100) and be mixed with each reference substance storing solution respectively.It is appropriate that precision measures each reference substance storing solution, be placed in same brown measuring bottle, be mixed with every 1ml respectively containing the mixing reference substance stock solution of jamaicin 0.4337mg, epiberberine 0.1052mg, coptis 0.0606mg, palmatine 0.1160mg, jateorrhizine 0.0721mg, (preserving with lower seal for 10 DEG C) for subsequent use.
(iii) need testing solution preparation: take coptis the water extracted immersing paste powdered sample and be about 50mg, accurately weighed, put in tool plug conical flask, add hydrochloric acid-methanol (1:100) 50ml, weighed weight, heat 15 minutes in 60 DEG C of water-baths, take out, ultrasonic process 20 minutes, lets cool, more weighed weight, the weight of less loss is supplied with hydrochloric acid-methanol (1:100), shake up, filter with miillpore filter (0.45 μm), to obtain final product.
(iv) liquid phase chromatography is measured:
Chromatographic condition: chromatographic column: Kromasil100-8C
18(250mm × 4.6mm); Mobile phase A: acetonitrile (1.5g lauryl sodium sulfate/500ml)-water (2:1) (phosphoric acid adjusts pH6.0), Mobile phase B: water (3.0g potassium dihydrogen phosphate/500ml), linear gradient elution program (0min:38%B, 55min:38%B, 65min:15%B, 70min:0%B); Determined wavelength: 345nm; Flow velocity: 1.0ml/min; Column temperature: 30 DEG C; Sample size: 10 μ l.
Measurement result is as shown in table 2.
Table 2: the active component content in Rhizoma Coptidis
(2) structure of the anti-shigella dysenteriae effect equivalence factor of the coptis:
1. shigella dysenteriae inoculation: essence gets 1.1mL shigella dysenteriae original bacteria liquid (forth generation), and be diluted to L.B. nutrient culture media the inoculation medium that cumulative volume is 44mL, namely original bacteria liquid expands 40 times, and shigella dysenteriae concentration is 1 × 10
6cFU/mL.2. under the environment of cleaning sterile, be in the glass ampoule bottles of 20mL, accurately add the L.B. nutrient culture media that 5mL has been vaccinated with shigella dysenteriae to each volume, then in each bottle, add the active component storing solution of different volumes respectively, be settled to 10mL, active component final concentration is made to be respectively 0.00mg/mL, 0.05mg/mL, 0.10mg/mL, 0.15mg/mL, 0.20mg/mL, 0.25mg/mL, add a cover bottle stopper, sealing, put into micro-calorimeter, " power against time (the power-time of shigella dysenteriae growth metabolism in each ampoule bottle of software detection followed the tracks of in the record carried by instrument, P-t) " curvilinear motion.The temperature of instrument is controlled in 37 DEG C, when all curves all return to baseline, terminates experiment.Corresponding data is derived by software, and by data importing Origin8.5 software (OriginLab company, the U.S.) mapping on curve.3. carry out bacteriostasis rate I% (Inhibitionratio) to calculate, formula is as follows: I%=(t
2– t
0)/t
0× 100%, wherein, t
0for the peak time of the second exponential phase without shigella dysenteriae growth metabolism during pharmaceutical intervention, t
2for the peak time of the second exponential phase of shigella dysenteriae growth metabolism during pharmaceutical intervention.According to the peak time t of the second exponential phase of shigella dysenteriae growth metabolism under each active component effect of the coptis
2, calculate the inhibiting rate I% that each composition of variable concentrations grows shigella dysenteriae.Bacteriostasis rate I% (y) and concentration C (x) are carried out linear relationship analysis, and the dose-effect relationship equation of both calculating, the results are shown in Table 3.
Table 3
The anti-shigella dysenteriae effect added by the dose-effect relationship equation of five kinds of active components and obtain the coptis adds and is worth, and adds and is worth the ratio of 0.9583, be the anti-shigella dysenteriae effect equivalence factor equation of this batch of coptis, i.e. formula I with the anti-shigella dysenteriae effect of control medicinal material composition:
EEF＝3.352*X
BER+1.293*X
EPI+6.635*X
COP+0.140*X
PAL+0.419*X
JAT-0.111。
(3) verify that the coptis anti-shigella dysenteriae effect equivalence factor is to the representativeness of its drug activity:
1. shigella dysenteriae inoculation: essence gets 1.1mL shigella dysenteriae original bacteria liquid (forth generation), and be diluted to L.B. nutrient culture media the inoculation medium that cumulative volume is 44mL, namely original bacteria liquid expands 40 times, and shigella dysenteriae concentration is 1 × 10
6cFU/mL.2. ampoule method is adopted, under the environment of cleaning sterile, be in the glass ampoule bottles of 20mL, accurately add the L.B. nutrient culture media that 5mL has been vaccinated with shigella dysenteriae to each volume, then in each bottle, add the coptis stock sample solution of different volumes respectively, be settled to 10mL, coptis water extract final concentration is made to be respectively 0.00mg/mL, 0.20mg/mL, 0.40mg/mL, 0.60mg/mL, 0.80mg/mL, 1.00mg/mL, add a cover bottle stopper, sealing, put into micro-calorimeter, the P-t curvilinear motion of shigella dysenteriae growth metabolism in each ampoule bottle of software detection followed the tracks of in the record carried by instrument.The temperature of instrument is controlled in 37 DEG C, when all curves all return to baseline, terminates experiment.3. with same concentration jamaicin for positive control, as investigation different batches coptis water extract, inhibiting concentration is produced, according to t to shigella dysenteriae using 0.20mg/mL
2, calculate different batches coptis sample to the inhibiting rate I of shigella dysenteriae
m%; And with the jamaicin inhibiting rate I of same concentrations
bER% is contrast, calculates the relative bacteriostasis rate RI of each batch of coptis water extract
m%, formula is RI
m%=I
m/ I
bER× 100%.4. the active component content value of each batch of coptis water extract is brought into formula I, calculate their EEF, the relative inhibition result of each batch sample and EEF are carried out correlation analysis, and result is as shown in table 4, both explanations have significant correlation, and EEF can represent Rhizoma Coptidis bacteriostatic activity preferably.
Table 4
The application of the anti-shigella dysenteriae effect equivalence factor EEF of the coptis:
Containing of the active component of each batch of Rhizoma Coptidis that the method according to abovementioned steps (1) is recorded
Value substitutes in formula I, and obtain the EEF of Rhizoma Coptidis, result is as shown in table 5:
Table 5
Then, according to formula II:D=D
target/ EEF, hypothetical target dosage D
targetit is 1 gram, then
The actual amount of Rhizoma Coptidis to be measured is as shown in table 6:
Table 6
What the present invention set up regulates the method empirical tests of coptis consumption can represent the anti-shigella dysenteriae activity of the coptis preferably based on anti-shigella dysenteriae effect equivalence factor.As the index of chemical composition analysis and biologically active aggregative weighted, it solves the problem of to run counter to result between the index by occurring during multiple index evaluation coptis quality and being difficult to evaluate medical material quanlity difference with the form of integrated quantification.Owing to being assigned with drug effect weight coefficient directly to each active component, coefficient is larger, then the representativeness of this composition to the coptis anti-shigella dysenteriae effect is stronger.When Clinical practice comments control coptis quality, can directly going to embody this batch of coptis it " effect " by detecting component content, not increasing the execution difficulty of quality standard, realizing simple, scientific and effective that traditional Chinese medicine quality comments control pattern.Meanwhile, effect equivalence factor to a certain degree can instruct clinical application, and the medicinal material that index is high suitably can reduce consumption when Clinical practice, and the medicinal material that index is low can increase quantity, to ensure the consistance of effect equivalent.To sum up, based on effect equivalent conforming coptis consumption adapting method, comprehensive quantification is made integrally to comment control Quality Evaluation of Chinese Medicinal and instruct clinical application to become possibility.
Method of the present invention also can carry out effect equivalence factor structure to different activities composition in the coptis; the consumption of different target dosage also can be adopted to adjust; these are only the example illustrated cited by summary of the invention; all coptiss made based on flesh and blood of the present invention batch, the simply change such as composition, target amount, all should drop in protection scope of the present invention.
Claims (3)
1. regulate a method for Rhizoma Coptidis consumption based on anti-shigella dysenteriae effect equivalence factor, said method comprising the steps of:
(1) measure the content of the jamaicin (BER) in described Rhizoma Coptidis, epiberberine (EPI), coptisine (COP), palmatine (PAL) and jateorrhizine (JAT), namely measure X respectively
bER, X
ePI, X
cOP, X
pAL, X
jAT;
(2) formula I is utilized to calculate the anti-shigella dysenteriae effect equivalence factor of Rhizoma Coptidis to be measured
Formula I:EEF
survey=3.352*X
bER+ 1.293*X
ePI+ 6.635*X
cOP+ 0.140*X
pAL+ 0.419*X
jAT-0.111
Described EEF
surveyfor the anti-shigella dysenteriae effect equivalence factor of Rhizoma Coptidis to be measured;
(3) formula II is utilized to calculate the actual amount of Rhizoma Coptidis
Formula II:D
survey=D
target/ EEF
survey; Wherein said D
surveyfor the actual amount of Rhizoma Coptidis to be measured, D
targetfor the target amount of Rhizoma Coptidis.
2. the method for claim 1, the described measurement wherein in step (1) adopts high performance liquid chromatography.
3. the method for claim 1, wherein said Rhizoma Coptidis is selected from ranunculaceae plant coptis CoptischinensisFranch., triangle leaf coptis CoptisdeltoideaC.Y.ChengetHsiao and Yun Lian CoptisteetaWall.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0928781A1 (en) * | 1997-12-30 | 1999-07-14 | Laboratoire Medidom S.A. | Process for the preparation of rhein and its diacyl derivatives |
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