CN108771728A - Inclined head seven is used to prepare the purposes of antifungal drug - Google Patents
Inclined head seven is used to prepare the purposes of antifungal drug Download PDFInfo
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- CN108771728A CN108771728A CN201810941414.2A CN201810941414A CN108771728A CN 108771728 A CN108771728 A CN 108771728A CN 201810941414 A CN201810941414 A CN 201810941414A CN 108771728 A CN108771728 A CN 108771728A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/10—Antimycotics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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Abstract
Inclined head seven disclosed by the invention is used to prepare the purposes of antifungal drug, belongs to pharmaceutical technology field.In vitro cell experiment shows, seven pairs of drug-resistant candida albicans 103 of inclined head, candida albicans SC5314, neogenesis cryptococcus, candida krusei, Candida parapsilosis, Candida glabrata, Candida tropicalis have different degrees of antimycotic even fungicidal action, therefore can inclined head seven be used as antifungal drug.The present invention is that inclined head seven opens new purposes, uses it for preparing antifungal drug, and new drug candidate is provided for fungal infection treatment.Clinically the wide and drug with Fungicidally active of anti-fungus spectra is considerably less at present, in the case where clinical fungi drug resistance is increasingly universal, drug-resistant intensity is on the rise, the characteristics of anti-fungus spectra of inclined head seven is wide and Fungicidally active, for the infection of clinical efficiently treatment fungi even drug resistance fungal, new therapy approach is provided.
Description
Technical field
The invention belongs to pharmaceutical technology fields, and in particular to inclined head seven is used to prepare the purposes of antifungal drug.
Background technology
Inclined head seven belongs to one kind of seven medicine of the Qinling Mountains, is Liliaceae Smilacina herbaceos perennial Smilacina japonica Smilacina
The drying root and rhizome of japonicaA.Gray and pipe flower Smilacina japonica Smilacina henryi (baker) .WangTang.With benefit
The effect of gas kidney-nourishing, dispelling wind and eliminating dampness and promoting blood circulation for regulating menstruation, can be used for treating treating rheumatic ostealgia, nervous headache, mastitis, menstruation not
The illnesss such as tune, carbuncle furuncle poison and traumatic injury.
But currently, the report of seven antifungic action of inclined head is had no both at home and abroad.
Invention content
The purpose of the present invention is to provide the purposes that inclined head seven is used to prepare antifungal drug.
The present invention is to be achieved through the following technical solutions：
The invention discloses application of the inclined head seven in preparing antifungal drug.
Preferably, select the drying root and rhizome of inclined head seven as the raw material for preparing antifungal drug.
Preferably, the fungi includes sensitive fungi bacterial strain and drug resistance fungal bacterial strain.
It is further preferred that the sensitive fungi bacterial strain includes：Sensitive candida albicans (SC5314), Candida parapsilosis
(22019) and neogenesis cryptococcus (32609)；The drug resistance fungal bacterial strain includes：Drug-resistant candida albicans (103), the drug resistance torrid zone are read
Pearl bacterium, natural drug resistance candida krusei (ATCC2340) and natural drug resistance Candida glabrata (ATCC2281).
It is that tablet, capsule is made in seven alcohol extract of inclined head and pharmaceutic adjuvant the invention discloses a kind of antifungal drug
Or injection.
Preferably, the preparation method of seven alcohol extract of inclined head is as follows：
The drying root and rhizome of inclined head seven is ground into powder, by 1g：The solid-liquid ratio of 20mL, with 75% ethyl alcohol condensing reflux
Extraction 2 hours, lets cool, is filtered to remove residue, and liquid is concentrated and dried and is made.
Preferably, the pharmaceutic adjuvant includes one or more of stabilizer, solubilizer, lubricant and disintegrant.
Compared with prior art, the present invention has technique effect beneficial below：
Inclined head seven disclosed by the invention is used to prepare the purposes of antifungal drug, shows inclined head by In vitro cell experiment
Seven pairs of drug-resistant candida albicans 103, candida albicans SC5314, neogenesis cryptococcus, candida krusei, Candida parapsilosis, smooth thought
Pearl bacterium, Candida tropicalis have different degrees of antimycotic even fungicidal action, therefore can be used as inclined head seven antimycotic
Drug opens new purposes for inclined head seven, uses it for preparing antifungal drug, and new time is provided for fungal infection treatment
Select drug.
Description of the drawings
Fig. 1 is the influence diagram of the various fungi growing states of seven pairs of inclined head of various concentration.
Specific implementation mode
With reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and
It is not to limit.
One, the effect of the different fungies of seven pairs of inclined head
1, reagent
Inclined head seven：It is purchased from Dare health chain drug store (the 4th chain store of Chinese scholartree bud).
Fluconazole：Sigma companies, lot number 036M4709V.
Dimethyl sulfoxide (DMSO)：Tianjin Tian Li chemical reagent Co., Ltd.
Each reagent is placed in -20 DEG C of preservations.Before experiment, drug taking-up is placed in 35 DEG C of incubators and is melted, is mixed well, respectively into
Row pharmacodynamics is examined.
2, bacterial strain
The bacterial strain used, including：
1) sensitive strain：Sensitive candida albicans (SC5314), Candida parapsilosis (22019) and neogenesis cryptococcus
(32609)；
2) antibody-resistant bacterium：Drug-resistant candida albicans (103), drug resistance Candida tropicalis, natural drug resistance candida krusei
(ATCC2340) and natural drug resistance Candida glabrata (ATCC2281).
The above bacterial strain is provided by Shanghai Changhai Hospital Mycology Lab, picks up from Changhai hospital different department clinical sample respectively,
And through morphology and biochemical identification.
All experiments draw plate activation, candida albicans, tropical beads with bacterial strain in husky fort glucose agar medium (SDA)
For the spherical bacterium such as bacterium, neogenesis cryptococcus after 30 DEG C are cultivated 2-3 days, picking monoclonal draws plate activation again respectively, takes second of institute
It obtains monoclonal and sets the inclined-planes SDA, saved backup in 4 DEG C after cultivating in aforementioned manners.
3, culture solution/base
1) 1640 liquid mediums of RPMI
RPMI1640 (Gibco BRL) 10g, NaHCO32.0g, morpholine propane sulfonic acid (Sigma) 34.5g
(0.165mol) adds tri-distilled water 900mL to dissolve, and 1mol/L NaOH tune pH to 7.0 (25 DEG C), tri-distilled water is settled to 1000mL,
0.22 μm of filtering with microporous membrane degerming, saves backup after packing in 4 DEG C.
2) husky fort agar glucose solid medium (SDA)
Peptone 10g, glucose 40g, agar 18g add tri-distilled water 900mL to dissolve, and 2mg/mL chloramphenicol solutions are added
50mL adjusts pH to 7.0, is settled to 1000mL with tri-distilled water, is saved backup in 4 DEG C after high pressure sterilization (121 DEG C, 15min).
3) YEPD culture solutions
Yeast extract 10g, peptone 20g, glucose 20g add tri-distilled water 900mL to dissolve, and 2mg/mL chloramphenicol water is added
Solution 50mL, tri-distilled water are settled to 1000mL, are saved backup in 4 DEG C after high pressure sterilization (121 DEG C, 15min).
4, instrument and equipment
Water isolation type electro-heating standing-temperature cultivator (Shanghai leap medical apparatus and instruments factory)；
THZ-82A Desk type constant-temperatureoscillator oscillators (Shanghai leap medical apparatus and instruments factory)；
SW-CT-IF types superpurgative working table (SuZhou Antai Air Tech Co., Ltd.).
5, seven extract of inclined head and Fluconazole stock solution
By the inclined head seven of purchase, it is ground into fine powder, weighs 5g, with the ethyl alcohol of 20 times of volumes (100mL) 75%, is condensed back to
Stream extraction 2 hours, lets cool, is filtered to remove residue, by liquid spin concentration to drying, takes 15mLDMSO dissolvings complete, prepares female
A concentration of 0.333g/mL of liquid, packing, -20 DEG C preserve for use.
Commercially available Fluconazole is dissolved with DMSO, a concentration of 6.4mg/mL is made into, is saved backup in -20 DEG C.
6, prepared by bacterium solution
Before experiment, with inoculation circle, each spherical fungi of picking is a small amount of respectively from the SDA culture mediums of 4 DEG C of preservations, is inoculated with respectively
To 1mL YEPD culture solutions, in 30 DEG C, 200rpm shaken cultivations activate 16h, fungi are made to be in later stage exponential phase of growth.Then
Each bacterium solution is added to respectively in 1mLYEPD culture solutions, is activated again in aforementioned manners, 16h is cultivated, with blood cell counting plate meter
Number, each bacterial concentration is adjusted separately to 1 × 10 with RPMI 1640 culture mediums3~5 × 103CFU/mL。
7, prepared by drug sensitive plate：
Each bacterial strain takes one piece of sterile 96 orifice plate respectively, adds 1640 fluid nutrient mediums of RPMI, 100 μ L to make in No. 1 hole of every row empty
White control；3~No. 12 holes respectively add the 100 μ L of bacterium solution for stating Fresh；No. 2 holes add 198 μ L of bacterium solution；No. 12 holes not drug containing,
Only plus 100 μ L of bacterium solution make Growth positive control.Mother liquid concentration is added to be the inclined of 0.3333g/mL and 6.4mg/mL in No. 2 holes of every row
First seven or each 2 μ L of Fluconazole.
10 grades of doubling dilutions are carried out to 2~No. 11 holes, it is respectively 3330 μ g/ to make final seven drug concentration of inclined head in each hole
mL、1665μg/mL、832.5μg/mL、416.3μg/mL、208.1μg/mL、104.1μg/mL、52.0μg/mL、26.0μg/mL、
The drug concentration of 13.0 μ g/mL and 6.5 μ g/mL, Fluconazole are respectively 64 μ g/mL, 32 μ g/mL, 16 μ g/mL, 8 μ g/mL, 4 μ g/
mL,2μg/mL,1μg/mL,0.5μg/mL,0.25μg/mL,0.125μg/mL.DMSO contents are below 1% in each hole.By each medicine
Quick plate is in 30 DEG C of insulating box cultures.
8, MIC value judges：
In 30 DEG C of insulating boxs, candida albicans is cultivated for 24 hours, neogenesis cryptococcus culture 72h, and observation experiment is as a result, and determine it
MIC value.It after observation, puts back in constant incubator, continues to cultivate.When the MIC value of drug is more than measured concentration range, press
Following methods are counted：When MIC value is higher than 64 μ g/mL of maximum concentration, be calculated as ">64μg/mL"；MIC value be minimum concentration or
It when below minimum concentration, does not make difference, is calculated as "≤0.125 μ g/mL ".The equal operation repetitive of above-mentioned experiment 3 times, works as MIC value
Can accurately repeat or only a poor concentration when just received, and using higher concentration as MIC value；When MIC value differs two concentration
When above, then need to test again, until meeting the requirements.
Experimental result is as shown in table 1 below：
The MIC value (μ g/mL) of 1 seven pairs of various fungies of inclined head of table
As it can be seen from table 1 partially head seven act on various fungies for 24 hours with after 48h, to Candida glabrata and neogenesis cryptococcus
MIC value is 416.3 μ g/mL, and the MIC value to candida albicans 103, candida albicans SC5314 and Candida parapsilosis is
832.5 μ g/mL, the MIC value to candida krusei is 1665.0 μ g/mL, is for 24 hours 416.3 μ to the MIC value of Candida tropicalis
G/mL, 48h are 832.5 μ g/mL.After drug effect 48h, to the MIC of candida krusei, Candida glabrata and neogenesis cryptococcus
32 μ g/mL of value difference, 8 μ g/mL and 8 μ g/mL, which show inclined heads seven to have antifungic action.
Two, the influence of the various fungi growing states of seven pairs of inclined head
Drug, bacterial strain and experiment material used are the same as above-mentioned shown.
By each bacterial strain with 1：100 are seeded to 1mLYEPD culture solutions, and in 30 DEG C, 200rpm shaken cultivations activate 16h, make true
Bacterium is in later stage exponential phase of growth.It takes in the bacterium solution to 1mLYEPD culture solutions, after activating 16h again in aforementioned manners, dilution is adjusted
Whole bacterial concentration is to 1 × 105~5 × 105CFU/mL takes the bacterium solution of 1mL Fresh to be positioned in glass tube, is separately added into partially
First seven or Fluconazole, drug grouping situation is：Blank group (not drug containing), Fluconazole (64 μ g/mL), (the 416.3 μ g/ of inclined head seven
ML), inclined head seven (832.5 μ g/mL), inclined head seven (1665 μ g/mL), inclined head seven (3330 μ g/mL).
For experimental result as shown in Figure 1, from figure 1 it appears that culture is for 24 hours with after 48h, the invisible spectro bacterium solution of blank group is muddy
It is turbid, and the inclined head seven of various concentration acts on invisible spectro bacterium solution and keeps relatively clarifying, and shows further seven various fungies of inclined head
The inhibiting effect of growth.
Three, the influence of seven pairs of various fungi survivings of inclined head
Drug, bacterial strain and experiment material used are the same as above-mentioned shown.
It is a small amount of from picking bacterium clone on the SDA culture mediums of 4 DEG C of preservations before experiment, 1mLYEPD culture solutions are seeded to, in 37
DEG C, 200rpm shaken cultivation 16h make fungi be in later stage exponential phase of growth.It is diluted with RPMI1640 culture solutions and adjusts bacterium solution
Concentration is to 1 × 105CFU/mL.Make its final concentration of 64mg/mL, 16mg/mL respectively at Fluconazole is added in above-mentioned bacterium solution, it is positive
Same volume is added in extract final concentration of 104.1mg/mL, 208.2mg/mL, 416.3mg/mL and 832.5mg/mL, blank control
Long-pending DMSO, DMSO contents are below 0.5% in all bacterium solutions.In 37 DEG C of incubators, with 200rpm shaken cultivations, respectively at
0h, for 24 hours, each sample is separately sampled, dilutes various concentration, and painting is laid on SDA tablets, and number clone, calculates each group after culture 48 hours
Colony survival number.
Experimental result is as shown in table 2 below：
2. inclined head seven of table acts on the surviving colonies number (CFU/ml) after 7 kinds of different fungies
As shown in Table 2, the inclined heads seven of a concentration of 416.3 μ g/mL and 832.5 μ g/mL act on 7 kinds of different fungi (candida albicans
SC5314, Candida parapsilosis, neogenesis cryptococcus, candida albicans 103, Candida tropicalis, candida krusei, Candida glabrata)
After for 24 hours, clump count is by about 105CFU/ml becomes 0, this shows the inclined head seven of a concentration of 416.3 μ g/mL and 832.5 μ g/mL to this
7 kinds of fungies all have preferable killing activity.
In conclusion the present invention shows inclined head seven pairs of drug-resistant candida albicans 103, candida albicans by In vitro cell experiment
SC5314, neogenesis cryptococcus, candida krusei, Candida parapsilosis, Candida glabrata, Candida tropicalis have different degrees of
Antimycotic even fungicidal action shows that inclined head seven has the characteristics that anti-fungus spectra is wide and Fungicidally active, therefore can will be inclined
First seven are used as antifungal drug.The present invention is that inclined head seven opens new purposes, uses it for preparing antifungal drug, is fungi
Treatment of infection provides new drug candidate, for the infection of clinical efficiently treatment fungi even drug resistance fungal, provides new control
Treatment approach.
Claims (7)
1. application of the inclined head seven in preparing antifungal drug.
2. application as described in claim 1, which is characterized in that select the drying root and rhizome of inclined head seven antimycotic as preparing
The raw material of drug.
3. application as described in claim 1, which is characterized in that the fungi includes sensitive fungi bacterial strain and drug resistance fungal bacterium
Strain.
4. application as claimed in claim 3, which is characterized in that the sensitive fungi bacterial strain includes：Sensitive candida albicans
(SC5314), Candida parapsilosis (22019) and neogenesis cryptococcus (32609)；The drug resistance fungal bacterial strain includes：Drug resistance is read in vain
Pearl bacterium (103), drug resistance Candida tropicalis, natural drug resistance candida krusei (ATCC2340) and natural drug resistance Candida glabrata
(ATCC2281)。
5. a kind of antifungal drug, which is characterized in that tablet, capsule or injection is made in seven alcohol extract of inclined head and pharmaceutic adjuvant
Agent.
6. antifungal drug according to claim 5, which is characterized in that the preparation method of seven alcohol extract of inclined head is as follows：
The drying root and rhizome of inclined head seven is ground into powder, by 1g：The solid-liquid ratio of 20mL is extracted with 75% ethyl alcohol condensing reflux
It 2 hours, lets cool, be filtered to remove residue, liquid is concentrated and dried and is made.
7. antifungal drug as claimed in claim 5, which is characterized in that the pharmaceutic adjuvant include stabilizer, solubilizer,
One or more of lubricant and disintegrant.
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| CN201810941414.2A CN108771728B (en) | 2018-08-17 | 2018-08-17 | Application of radix pseudostellariae to preparation of antifungal drugs |
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| CN201810941414.2A CN108771728B (en) | 2018-08-17 | 2018-08-17 | Application of radix pseudostellariae to preparation of antifungal drugs |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109700859A (en) * | 2019-03-13 | 2019-05-03 | 陕西科技大学 | Polygonum cill is preparing the application in antifungal drug and its synergist |
| CN113082150A (en) * | 2021-04-30 | 2021-07-09 | 陕西科技大学 | Application of effective part of deer medicine in preparing antifungal medicine and synergist thereof |
-
2018
- 2018-08-17 CN CN201810941414.2A patent/CN108771728B/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109700859A (en) * | 2019-03-13 | 2019-05-03 | 陕西科技大学 | Polygonum cill is preparing the application in antifungal drug and its synergist |
| CN109700859B (en) * | 2019-03-13 | 2021-11-16 | 陕西科技大学 | Application of radix notoginseng cinnabaris in preparation of antifungal drugs and synergists thereof |
| CN113082150A (en) * | 2021-04-30 | 2021-07-09 | 陕西科技大学 | Application of effective part of deer medicine in preparing antifungal medicine and synergist thereof |
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