CN106266264A - A kind of pharmaceutical composition of anti-mycobacterium tuberculosis and preparation method thereof - Google Patents
A kind of pharmaceutical composition of anti-mycobacterium tuberculosis and preparation method thereof Download PDFInfo
- Publication number
- CN106266264A CN106266264A CN201510313738.8A CN201510313738A CN106266264A CN 106266264 A CN106266264 A CN 106266264A CN 201510313738 A CN201510313738 A CN 201510313738A CN 106266264 A CN106266264 A CN 106266264A
- Authority
- CN
- China
- Prior art keywords
- callicarpa nudiflora
- pharmaceutical composition
- preparation
- extract
- petroleum ether
- 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.)
- Pending
Links
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of Callicarpa nudiflora medical composition and its use, described pharmaceutical composition contains 1 part of Callicarpa nudiflora petroleum ether extraction effective site B and 3-5 part Callicarpa nudiflora extract C, and gained pharmaceutical composition of the present invention has stronger anti-mycobacterium tuberculosis effect.
Description
Technical field
The present invention relates to field of medicaments, be specifically related to a kind of pharmaceutical composition with anti-mycobacterium tuberculosis effect and preparation method thereof.
Background technology
Callicarpa nudiflora (Callicarpa nudiflora Hook.et Arn.) is Verenaceae Callicarpa plant, originate from China Hainan,
Guangxi, Guangdong, also there is distribution on the ground such as India, Vietnam, Malaysia, and its root, leaf can be used as medicine, and have astringing to arrest bleeding, antiinflammatory
The effects such as removing toxic substances.Effect to the anti-mycobacterium tuberculosis of Callicarpa nudiflora in nearly 2 years has a small amount of research, as " Callicarpa nudiflora is to being clinically separated
The In Vitro Bacteriostasis of mycobacterium tuberculosis " (China's Chinese medicine magazine, 2014,29 (11): 3630-3632), " Callicarpa nudiflora receipts
Hold back hemostasis and the antibacterial action of mycobacterium tuberculosis studied " (Hunan Normal University, Master's thesis in 2014), preliminary study
To the effect to mycobacterium tuberculosis of the Callicarpa nudiflora dry extract, but its material base the distinctest.
Summary of the invention
It is an object of the invention to provide a kind of Callicarpa nudiflora pharmaceutical composition, be achieved through the following technical solutions:
A kind of pharmaceutical composition, it is characterised in that be 1 part of Callicarpa nudiflora petroleum ether extraction effective site B and 3-5 containing weight portion
Part Callicarpa nudiflora extract C, the preparation method of wherein said Callicarpa nudiflora petroleum ether extraction effective site B is:
(1) take Callicarpa nudiflora medical material, be ground into coarse powder, add 90%-95% alcohol heating reflux and extract 1-3 time, obtain extracting solution,
After filtration, gained filtrate is condensed into extractum A,
(2) step (1) gained extractum A is dispersed in water, adds petroleum ether extraction, removes solvent, concentrates, and is dried
Obtain petroleum ether extraction effective site B,
The preparation method of described Callicarpa nudiflora extract C is:
(1) taking Callicarpa nudiflora chopping, extracting in water, Extracting temperature 85 DEG C-95 DEG C, extracting solution filters, and filtrate is dense to relatively
Density 1.05-1.15 (60 DEG C),
(2) in step (1) gained filtrate, add ethanol to alcohol content 40%-50%, stand 10-14 hour, filter,
(3) in step (2) gained filtrate, add ethanol to alcohol content 60%-70%, stand 10-14 hour, filter, filtrate
Concentrate, be dried to obtain Callicarpa nudiflora extract C.
Preferred as such scheme, the preparation method step (1) of described petroleum ether extraction effective site B is: take nakedflower purple
Pearl medical material, is ground into coarse powder, adds 95% alcohol heating reflux and extracts 3 times, obtains extracting solution, and after filtration, gained filtrate decompression is dense
Contract to obtain extractum A.
Preferred as such scheme, the preparation method step (1) of described Callicarpa nudiflora extract C is: takes Callicarpa nudiflora and cuts
Broken, add 6-8 times amount water extraction 1-3 time, Extracting temperature 85 DEG C-95 DEG C, extracting solution filters, and filtrate is dense to relative density 1.05
-1.15 (60 DEG C)；In the preparation method step (2) of described Callicarpa nudiflora extract C, alcohol content is 45%；Described nakedflower is purple
In the preparation method step (3) of pearl extract C, alcohol content is 65%.
Described in pharmaceutical composition of the present invention, the weight portion of Callicarpa nudiflora extract C is preferably 4 parts.
It is a further object to provide described pharmaceutical composition preparation have in anti-mycobacterium tuberculosis drugs with function purposes and
Purposes in preparation treatment tubercular drugs.
Pharmaceutical composition of the present invention, containing the described pharmaceutical composition of any of the above-described item of effective dose and pharmaceutically can be able to connect
The adjuvant being subject to, is prepared as pharmaceutical preparation.
The chemical composition of Callicarpa nudiflora have flavone and glycoside, triterpene and glycoside, diterpene, phenethyl alcohol glycosides, phenolic acids, steroid,
Volatile oil etc., the most therefrom more than 100 compounds of isolated.But it is existing to Callicarpa nudiflora composition and tuberculosis branch
The research of bacillus effect is the weakest, and existing research conclusion speculates that Main Function composition is flavonoid.It is known that Chinese medicine ingredients
Complexity, typically plays tuberculosis effect, going out of multi-drug resistant tubercule bacillus by multicomponent, Mutiple Targets, overall coordination effect
Now make to control tuberculosis and remain a global problem being badly in need of solving.Inventor on the basis of early-stage Study, Antituberculous
Bacillus active ingredient group has done in-depth study, by using solvent (water, methanol, ethanol, acetone etc.) to extract, in conjunction with big
Hole resin or activated carbon or ceramic membrane or extraction etc. separation method, combine the analytical technology such as mass spectrum, liquid phase, finally draw this
Bright technical scheme, its experiment effect is shown in detailed description of the invention with the contrast such as comparative example, positive control drug.
Extract at low temperature (low-temperature dynamic extraction) gained filtrate afterwards adds the ethanol of variable concentrations and carries out the naked of with different levels precipitate with ethanol acquisition
Flower Folium Callicarpae Formosanae extract, its tuberculosis effect is obviously improved, and on the one hand uses low-temperature dynamic to extract, and retains as far as possible and is oxidized easily
Flavonoid, the active materials such as phenethyl alcohol glycosides, on the other hand the ethanol precipitate with ethanol of variable concentrations is conducive to removing more fully naked
The non-active ingredient such as phlegmatic temperament, polysaccharide, pigment and Antituberculous effect may produce the material of antagonism in flower Folium Callicarpae Formosanae crude extract,
Enrichment and purification effective active composition further, improves the biological activity of unit extract.
Beneficial effects of the present invention: (1) obtains the pharmaceutical composition that Killing Mycobacterium Tuberculosis effect is stronger, (2) preparation method is grasped
Make simplicity.
Detailed description of the invention
Hereinafter will describe in detail the most by way of example, inventor obtains corresponding embodiment and comparative example by Different Preparation,
And record the MIC value of embodiment, comparative example, positive control drug, refer to table 1.
Inventor prepares embodiment by following technique:
1, the preparation of Callicarpa nudiflora petroleum ether extraction effective site B:
(1) take Callicarpa nudiflora medical material, be ground into coarse powder, add 90%-95% alcohol heating reflux and extract 1-3 time, obtain extracting solution,
After filtration, gained filtrate is condensed into extractum A,
(2) step (1) gained extractum A is dispersed in water, adds petroleum ether extraction, removes solvent, concentrates, and is dried
Obtain petroleum ether extraction effective site B,
2, the preparation of Callicarpa nudiflora extract C:
(1) taking Callicarpa nudiflora chopping, add 6-8 times of water extraction 1-3 time, Extracting temperature 85 DEG C-95 DEG C, extracting solution filters, filter
Liquid is dense to relative density 1.05-1.15 (60 DEG C),
(2) in step (1) gained filtrate, add ethanol to alcohol content 40%-50%, stand 10-14 hour, filter,
(3) in step (2) gained filtrate, add ethanol to alcohol content 60%-70%, stand 10-14 hour, filter, filtrate
Concentrate, be dried to obtain Callicarpa nudiflora extract C.
3, Callicarpa nudiflora petroleum ether extraction effective site B and Callicarpa nudiflora extract C mixed with the weight ratio of 1:3-5 and get final product.
Each embodiment, comparative example, positive control drug are as follows to the experimental technique of suppression mycobacterium tuberculosis:
1. material
1.1 bacterial strains: mycobacterium tuberculosis reference culture MTB H37Rv (offer of Tuberculosis Control Institute of Hunan Province)
1.2 culture medium and antibacterials
Basal medium is modified Russell medium；Antibacterials are each embodiment, comparative example, positive control drug.
1.3 reagent: Middlebrook 7H9 meat soup OADC enrichment liquid is purchased from BD Difco company；Positive control drug isoniazid, profit
Good fortune is flat, ethambutol is purchased from sigma company；DMSO deionized water, tween 80 are purchased from the traditional Chinese medicines group limited public affairs of chemical reagent
Department, is chemical analysis pure；96 microwell plates (Corning, the U.S.)；Alamar blue (Serotec, Britain)；7H9 (Difco,
The U.S.)；OADC (BD, the U.S.).
2. experiment and method
The preparation of 2.1 bacterium colonies: make basal liquid by tubercule bacillus inoculated and cultured according to improvement Roche method；
The preparation of 2.2 original liquid concentrations: take about 16g gained of the present invention pharmaceutical composition DMSO and dissolve, with 0.22um after fully dissolving
Membrane filtration, degerming, standby.Comparative example gained sample herewith method is prepared.Isoniazid 5g, ethambutol 5g are abundant with water respectively
With 0.22um membrane filtration after dissolving, degerming, standby.Rifampicin 5g with DMSO dissolve after with 0.22um membrane filtration,
Degerming, standby.
2.3 experiment
The sterile deionized water of 200ul is joined the hole of the outer perimeter of all 96 aseptic orifice plates, hatches period with minimizing as far as possible
The evaporation of the medium in instrument connection.With aseptic Middlebrook 7H9 broth dilution Callicarpa nudiflora stock solution and other three kinds of positives
(Callicarpa nudiflora is 32mg/ml to 4 times of final detectable concentrations of greatest hope of medicine, and isoniazid is 1.6 μ g/ml, and ethambutol is
40 μ g/ml, rifampicin is 8 μ g/ml) add 100 μ l in detection hole, then medicine is carried out 2 times of serial dilutions and dilutes respectively
To 1/2,1/4,1/8,1/16,1/32,1/64.Each drug moiety concentration repeats 2 holes.Choose and grow on Russell medium
The fresh colony in 2-3 week, with glass bead mill bacterium, (McFarland 1 is equivalent to (3-6) to be diluted to 1 Maxwell concentration
×107CFU/ml ", then dilute backward microwell plate addition 100ul bacterium solution with 1:25, the final concentration of bacterium solution is about 106CFU/ml。
Each bacterial strain sets 1 not growth control hole (positive control) of drug containing and comprise only 7H9 not drug containing and the bacteria control of bacterium solution
Hole (negative control), each bacterial strain is parallel carries out 3 tests.96 orifice plates add in growth control hole after hatching 5d in 37 DEG C
The mixed liquor of 20ulAlamar blue and 50ulTweerr80 (5%), hatches 24h for 37 DEG C, if color becomes pink colour from blueness,
In each Experimental agents hole, then add Alamar blue and the Tweerr80 mixed liquor of above-mentioned amount, hatch 24h for 37 DEG C and record each hole
Color, blue Kong Weiwu growth, pink hole is for there being growth.As aubergine hole occurs, then continue to cultivate 24h at 37 DEG C,
If not becoming pink and its blue hole being connected being still blueness, then it is recorded as growth.MIC is defined as stoping color change
The lowest concentration of drug of (becoming pink from blueness).
Said method parallel test, applies identical bacterium solution and concentration and medicine with in a few days operating, carries out 3 times.The MIC of report
Result is 2 times and more than 2 times identical MIC value.Experimental result see table 1.
Table 1
Each embodiment, blank group, positive control drug group the experimental technique of anastalsis as follows:
90 healthy Kunming white mice are divided at random 9 groups (often group 10, male and female half and half), be divided into experimental group (embodiment 1-7),
Positive control drug group (YUNNAN BAIYAO group), blank group, successive administration 7 days, experimental group and the dosage of positive control drug group
0.5g/kg, gavage volume 0.2mL/10g body weight, blank group gives equal-volume normal saline, after last day is administered 45min,
Measure the mice bleeding time (BT) with cutting tail method, survey clotting time (CT) by capillary glass-tube method, evaluate drug hemostasis effect.Real
Test result and see table 2.
The impact (mean ± standard deviation, N=10) on mice anastalsis of table 2 present composition
| Group | Dosage | BT (second) | CT (second) |
| Blank group | 20.56±2.32 | 66.91±4.85 | |
| Embodiment 1 | 0.5g/kg | 14.27±1.35** | 45.08±3.16** |
| Embodiment 2 | 0.5g/kg | 14.02±1.57** | 46.12±3.41** |
| Embodiment 3 | 0.5g/kg | 13.81±1.31** | 43.59±4.07** |
| Embodiment 4 | 0.5g/kg | 13.55±1.42** | 43.18±3.76** |
| Embodiment 5 | 0.5g/kg | 13.28±1.25** | 42.71±3.93** |
| Embodiment 6 | 0.5g/kg | 14.09±1.44** | 43.46±3.25** |
| Embodiment 7 | 0.5g/kg | 13.62±1.18** | 42.15±3.52** |
| YUNNAN BAIYAO group | 0.5g/kg | 13.98±2.03** | 46.15±3.87** |
* P < 0.05, * * P < 0.01 compares with blank group.
Table 1 shows that the anti-mycobacterium tuberculosis effect of gained pharmaceutical composition of the present invention is relatively strong, compared with prior art, has significance difference
Different, can be used for preparing in the medicine of anti-mycobacterium tuberculosis effect, and gained pharmaceutical composition has astringing to arrest bleeding effect (being shown in Table 2),
The comprehensive function of astringing to arrest bleeding and anti-mycobacterium tuberculosis can be given full play to, be used for treating tuberculosis.
Claims (9)
1. a pharmaceutical composition, it is characterised in that be 1 part of Callicarpa nudiflora petroleum ether extraction effective site B and 3-5 part containing weight portion
Callicarpa nudiflora extract C, the preparation method of wherein said Callicarpa nudiflora petroleum ether extraction effective site B is:
(1) take Callicarpa nudiflora medical material, be ground into coarse powder, add 90%-95% alcohol heating reflux and extract 1-3 time, obtain extracting solution, mistake
After filter, gained filtrate is condensed into extractum A,
(2) step (1) gained extractum A is dispersed in water, adds petroleum ether extraction, removes solvent, concentrates, is dried to obtain
Petroleum ether extraction effective site B,
The preparation method of described Callicarpa nudiflora extract C is:
(1) taking Callicarpa nudiflora chopping, extracting in water, Extracting temperature 85 DEG C-95 DEG C, extracting solution filters, and filtrate is dense to relative density
1.05-1.15 (60 DEG C),
(2) in step (1) gained filtrate, add ethanol to alcohol content 40%-50%, stand 10-14 hour, filter,
(3) adding ethanol in step (2) gained filtrate to alcohol content 60%-70%, stand 10-14 hour, filter, filtrate concentrates,
It is dried to obtain Callicarpa nudiflora extract C.
Pharmaceutical composition the most according to claim 1, it is characterised in that the preparation side of described petroleum ether extraction effective site B
Method step (1) is: takes Callicarpa nudiflora medical material, is ground into coarse powder, adds 95% alcohol heating reflux and extracts 3 times, obtains extracting solution,
After filtration, gained filtrate reduced in volume obtains extractum A.
Pharmaceutical composition the most according to claim 1, it is characterised in that the preparation method step of described Callicarpa nudiflora extract C
Suddenly (1) is: takes Callicarpa nudiflora chopping, adds 6-8 times amount water extraction 1-3 time, Extracting temperature 85 DEG C-95 DEG C, and extracting solution filters,
Filtrate is dense to relative density 1.05-1.15 (60 DEG C).
Pharmaceutical composition the most according to claim 1, it is characterised in that the preparation method step of described Callicarpa nudiflora extract C
Suddenly in (2), alcohol content is 45%.
Pharmaceutical composition the most according to claim 1, it is characterised in that the preparation method step of described Callicarpa nudiflora extract C
Suddenly in (3), alcohol content is 65%.
Pharmaceutical composition the most according to claim 1, it is characterised in that the weight portion of described Callicarpa nudiflora extract C is 4
Part.
7. pharmaceutical composition described in any one of claim 1-6 has the application in anti-mycobacterium tuberculosis drugs with function in preparation.
8. the application in preparation treatment tubercular drugs of the pharmaceutical composition described in any one of claim 1-6.
Pharmaceutical composition the most according to claim 1, it is characterised in that described in any one of claim 1-6 containing effective dose
Pharmaceutical composition and pharmaceutically acceptable adjuvant, be prepared as pharmaceutical preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510313738.8A CN106266264A (en) | 2015-06-10 | 2015-06-10 | A kind of pharmaceutical composition of anti-mycobacterium tuberculosis and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510313738.8A CN106266264A (en) | 2015-06-10 | 2015-06-10 | A kind of pharmaceutical composition of anti-mycobacterium tuberculosis and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106266264A true CN106266264A (en) | 2017-01-04 |
Family
ID=57659164
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510313738.8A Pending CN106266264A (en) | 2015-06-10 | 2015-06-10 | A kind of pharmaceutical composition of anti-mycobacterium tuberculosis and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106266264A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234159A (en) * | 2007-01-29 | 2008-08-06 | 陈晓坚 | Pharmaceutical combination with synergistic reaction |
| CN103479798A (en) * | 2012-06-13 | 2014-01-01 | 九芝堂股份有限公司 | Traditional Chinese medicine for resisting mycobacterium trberculosis and preparation method thereof |
-
2015
- 2015-06-10 CN CN201510313738.8A patent/CN106266264A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234159A (en) * | 2007-01-29 | 2008-08-06 | 陈晓坚 | Pharmaceutical combination with synergistic reaction |
| CN103479798A (en) * | 2012-06-13 | 2014-01-01 | 九芝堂股份有限公司 | Traditional Chinese medicine for resisting mycobacterium trberculosis and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tambe et al. | Phytochemical screening and antibacterial activity of Syzygium cumini (L.)(Myrtaceae) leaves extracts | |
| Molina-Salinas et al. | Evaluation of the flora of Northern Mexico for in vitro antimicrobial and antituberculosis activity | |
| Neelavathi et al. | Antibacterial activities of aqueous and ethanolic extracts of Terminalia catappa leaves and bark against some pathogenic bacteria | |
| Muhammad et al. | Phytochemical screening and antimicrobial activities of Terminalia catappa, leaf extracts | |
| Jafarı-sales et al. | Antibacterial effect of methanolic extract of saffron petal (Crocus sativus L.) on some standard gram positive and gram negative pathogenic bacteria in vitro | |
| Al-Abd et al. | Phytochemical constituents, antioxidant and antibacterial activities of methanolic extract of Ardisia elliptica | |
| Rajamohan et al. | Antioxidant, Antimicrobial activities and GC-MS analysis of Calotropis gigantea white flowers | |
| Ogidi et al. | Determination of Phytoconstituents and Antimicrobial activities of aqueous and methanol extracts of neem (Azadirachta indica) leaves | |
| CN106266265A (en) | A kind of Callicarpa nudiflora Chinese medicine composition and its production and use | |
| Geetha et al. | In vitro evaluation of anti mycotic activity of ethanolic extract of Glycyrrhiza glabra | |
| Sugandhi et al. | Antimicrobial activity of Mentha arvensis L.(Lamiaceae) | |
| CN106266263A (en) | A kind of Chinese medicine composition and its production and use | |
| Suliman et al. | Chemical composition and antibacterial activity of crude extracts from Sudanese medicinal plant Adansonia digitata L | |
| Gowrish et al. | In vitro antioxidant and antitubercular activity of Leucas marrubioides Desf. root extracts | |
| Mbakwem-Aniebo et al. | Effects of Ficus exasperata Vahl on common dermatophytes and causative agent of Pityriasis Versicolor in rivers state, Nigeria | |
| Sunday et al. | Antibacterial effect of Vernonia amygdalina and Ocimum gratissimum on some bacteria that cause opportunistic infections in HIV/AIDS patients | |
| Cruz et al. | Antimicrobial activity of essential oils and ethanolic extracts of three species of laurel (Litsea spp.) from Guatemala | |
| Ruma et al. | Determination of secondary metabolites and antibacterial property of extract from the leaves of Stachytarpheta jamaicensis (L.) Vahl | |
| Lai et al. | Evaluation of antibacterial and antifungal activities of Persicaria chinensis leaves | |
| CN106266264A (en) | A kind of pharmaceutical composition of anti-mycobacterium tuberculosis and preparation method thereof | |
| Guzel et al. | Evaluation of antimicrobial activity of five Vincetoxicum taxa growing in Turkey | |
| Aska et al. | Quantitative phytochemical analysis and Anti-tuberculosis activity of some selected medicinal plants in some Northern parts of Bauchi state, Nigeria | |
| Banu et al. | Screening of bioactive compounds and antimicrobial effect of Orithazh thamarai chooranam | |
| Ghani et al. | Determination and identification of phytochemical properties of medicago sativa l.(alfalfa) leaf, stem and root extracts against various pathogens | |
| Al-Jeboury | In vitro antibacterial activity of Eucalyptus microtheca and Cinnamomum zeylanicum crude extracts against Pseudomonas aeruginosa isolates |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170104 |
|
| RJ01 | Rejection of invention patent application after publication |