CN108186700A - A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals - Google Patents

A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals Download PDF

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CN108186700A
CN108186700A CN201810237327.9A CN201810237327A CN108186700A CN 108186700 A CN108186700 A CN 108186700A CN 201810237327 A CN201810237327 A CN 201810237327A CN 108186700 A CN108186700 A CN 108186700A
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phenolic acids
total phenolic
water
water cress
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黄正明
刘青川
曹文斌
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    • A61K2236/50—Methods involving additional extraction steps
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    • 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
    • A61K2236/55—Liquid-liquid separation; Phase separation

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Abstract

The invention discloses a kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that：With total phenolic acids of water cress extract bulk pharmaceutical chemicals, which contains total phenolics 55 80%.Total phenolic acids of water cress experiment in vitro finds that it has stronger HIV-resistant activity to HIV；Experiment in vivo finds that it can significantly improve CD4 +The percentage of T cell improves CD4 +/CD8 +Value, by improving the immunosuppressive action of body, so as to play antivirus action；Total phenolic acids of water cress has good inhibiting effect to the activity of hiv reverse transcriptase.Total phenolic acids of water cress inside and outside is respectively provided with the effect of AntiHIV1 RT activity, and function and effect are clear and definite, and is by the way that the activity of hiv reverse transcriptase is inhibited to have the function that AntiHIV1 RT activity, and the mechanism of action is more clear and definite, has the potential quality of exploitation Clinical practice.

Description

It is a kind of to be used to prepare anti-hiv drug preparation by bulk pharmaceutical chemicals of total phenolic acids of water cress Purposes
Technical field
The invention belongs to the separation of plant active component, technical field of purification, and Chinese celery active component total phenol acid content can be made big In 50%, reach national 5 class Chinese medicine material medicine standards；And it is related to new as the anti-AIDS of the pharmaceutical preparation of bulk pharmaceutical chemicals using the ingredient Purposes.
Background technology
The present inventor team has studied a kind of preparation process of total phenolic acids of water cress and the medicine using the total phenolics as active constituent Object preparation and application (patent No. 2006100838103) and a kind of total phenolic acids of water cress preparation process and using the ingredient as bulk pharmaceutical chemicals Pharmaceutical preparation and hepatopathy purposes (patent No. 2006101607233), extraction process to total phenolic acids of water cress and application carry out Preliminary exploration.Common extraction and separation process is：Alcohol extracting-water sinks-ethyl acetate extraction method or acid-base method, total phenolics are obtained, this A little method loss of effective components are big or ingredient is corrupted such that total phenolics yield is low, of high cost.Therefore further research one is needed New easy, the efficient extraction separation method of kind, and suitable for industrialized production.Meanwhile in face of this medical field of AIDS Problem, need to explore the pharmaceutical preparation new application for preparing anti AIDS virus using total phenolic acids of water cress as bulk pharmaceutical chemicals.
Invention content
Present invention aims at proposition one kind anti-hiv drug preparation is used to prepare by bulk pharmaceutical chemicals of total phenolic acids of water cress Purposes.The extraction process of total phenolic acids of water cress is optimized simultaneously.
The present invention seeks to what is be achieved through the following technical solutions：One kind is used to prepare anti-using total phenolic acids of water cress as bulk pharmaceutical chemicals The purposes of AIDS virus pharmaceutical preparation, it is characterised in that：With total phenolic acids of water cress extract bulk pharmaceutical chemicals, which contains total phenolics 55-80%.
A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that： Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals, using addition cosolvent such as Tween-80 or using water-miscible Propylene glycol, polyethylene glycol-400 dissolving or using with the methods of organic base forming salt water soluble preparation being made.
A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that： Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals, the total phenolic acids of water cress extraction containing 0.5%-99.5% in component The drug excipient of raw material medicine and 99.5%-0.5%.
A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that： Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals, the total phenolic acids of water cress extract containing 1%-60% is former in component Expect medicine and the drug excipient of 99%-40%.
A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that： Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals is oral preparation or parenteral dosage form.
A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that： Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals, the oral preparation selected from tablet, pill, capsule, Granula, suspension, dripping pill, oral liquid any one of are worked as.
A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that： Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals, the parenteral dosage form selected from injection, aerosol, Suppository or subcutaneous administration dosage form any one of are worked as.
The excipient, one or several groups including solvent, disintegrant, corrigent, preservative, colorant and adhesive It closes.The drug of other adapteds is also included in the pharmaceutical excipient.
The drug of other adapteds included in the pharmaceutical excipient refers to using the total phenolic acids of water cress of effective dose as medicine Raw material, compatibility other allowed the Chinese medicine or chemicals that share.
The preparation method of total phenolic acids of water cress bulk pharmaceutical chemicals is：
Dry Chinese celery herb is ground into coarse powder, the hydrophilic solvent heating and refluxing extraction or cold soaking measured with 10-30 times Diafiltration extraction, heat carries 2-3 times, 1-3 hours each, and extracting solution then is concentrated under reduced pressure into proportion under the conditions of less than 60 DEG C The medicinal extract of (1.15-1.30 70 DEG C)；Primary 4-6 times of hot water of dose is added in, is dissolved by several times, stands 24 hours, is filtered, is gathered on filtrate Amide is the column chromatography of carrier, is rinsed with water and uses water：Alcohol ratio is 50：50-5：95 mixed solvent elution, collects water-alcohol Eluent is concentrated under reduced pressure under conditions of less than 60 DEG C, is dried in vacuo to obtain total phenolic acids of water cress extract.
The preparation method of total phenolic acids of water cress bulk pharmaceutical chemicals can also be：Dry Chinese celery herb is ground into coarse powder, adds 10 times 70% ethyl alcohol heating and refluxing extraction of amount 2 hours totally 3 times, merges extracting solution, lets cool, filters, then by extracting solution in less than 60 The medicinal extract of proportion 1.15-1.30 (70 DEG C) is concentrated under reduced pressure under the conditions of DEG C；Primary 6 times of hot water of dose are added in, are dissolved by several times, are stood 24 hours, filtering, filtrate was added on 5 times of the crude drug amount that is equivalent to, and on processed good polyamide column, was rinsed with water to efflux in light It after yellow, is eluted with 70% alcohol, collects ethanol eluate until eluent is in faint yellow, depressurized under conditions of less than 60 DEG C Concentration, is dried in vacuo to obtain total phenolic acids of water cress extract.
The present invention has the advantages that：Total phenolic acids of water cress experiment in vitro has stronger HIV-resistant activity to HIV；Body Interior experiment can significantly improve CD4 +The percentage of T cell improves CD4 +/CD8 +Value, by improving the immunosuppressive action of body, from And play antivirus action；Total phenolic acids of water cress has good inhibiting effect to the activity of hiv reverse transcriptase.Total phenolic acids of water cress body The inside and outside effect for being respectively provided with AntiHIV1 RT activity, function and effect are clear and definite, and are by the way that the activity of hiv reverse transcriptase is inhibited to reach AntiHIV1 RT activity Effect, the mechanism of action is more clear and definite, has the potential quality of exploitation Clinical practice.Using total phenolic acids of water cress as the pharmaceutical preparation energy of bulk pharmaceutical chemicals It is enough in anti-AIDS purposes.
Specific embodiment
Examples below is used to further illustrate the present invention, but be not intended to limit rights protection scope of the present invention.
Experimental example 1：Total phenolic acids of water cress is in vitro to the inhibiting effect of AIDS virus
1. experiment material
1.1 experimental cell TZM-B1 cells, from HeLa cell lines, which can infect the HIV of various preferendums, monkey Immunodeficiency virus (SIV) and monkey-human immunodeficiency virus (sHIV) etc., and primary viral and pseudovirus can infect.
1.2 Viral experiment adapted strain SF33 viruses, are provided by Chinese Center for Disease Control and Prevention.
1.3 reagent D MEM culture mediums, CORNING；FBS fetal calf serums, Gibco；Glutamine (G) L-Glutamine, Gibco；Mycillin, Gibco；BriteliteTMplus Reporter Gene Assay System PerkinElmer 6066769。
1.4 laboratory apparatus carbon dioxide incubators, Thermo scientific HERAcell 150i；Optical microphotograph Mirror, OLYMPUS CKX41；Blood-counter system, NIHON KOHDEN MEK-6318K；Luminescence of cell detector, PerkinElmer 1420 Multilabel Counter。
2 test methods
2.1 fishplate bars are inoculated with TZM-bl cells in 96 orifice plates, and 104/ hole is placed in 37 DEG C, 5%CO2It is cultivated in incubator, Start to test afterwards for 24 hours.
2.2 make up a prescription administration total phenolic acids of water cress and T-20 be respectively diluted to 7 dilutions with complete DMEM culture mediums； SF33 Virus Samples are diluted to 2000TICD with the DEAE culture solutions containing DEAE-dextran50The end of/ml, DEAE-dextran A concentration of 15 μ g/ml.Liquid suction in 96 orifice plates is abandoned, sequentially adds the drug diluted, 100 μ l/ holes, while it is right to set virus According to group (VC) and cell controls group (CC), the complete DMEM culture solutions of 100 μ l, each medicine are added in virus control and cell control well Each dilution of object sets 2 multiple holes, the virus diluted is added in 96 orifice plates, 100 μ l/ hole (i.e. 200TICD50/ hole), carefully Born of the same parents' control wells add in the complete DMEM culture solutions of 100 μ l.
96 well culture plates are sealed after the administration of 2.3 viral diagnosis, 37 DEG C is placed in and cultivates 48 hours, then take out 96 holes 100 μ l are sucked out per hole for plate, and 100 μ l of addition, which shine, detects liquid, and room temperature avoid light place 2min draws 150 μ l supernatants and is transferred to black In 96 orifice plates, luminous value is detected in luminescence of cell detector, IC is calculated according to Reed and Muench methods50。
3. experimental result
The influence of 3.1 total phenolic acids of water cress intracellular inhibition of HIV to TZM-B1
After TZM-B1 cell infection SF33 viruses, virus control group luminescence of cell value is significantly raised, shows cell infection disease Poison success.After 48h is administered, compared with virus control group, total phenolic acids of water cress and each dosage group virus luminous value of positive control drug are equal It is decreased obviously, in dose-effect relationship, there is significant difference (P ＜ 0.01), as a result there is repeatability, the results are shown in Table 1.
The influence (n=2) of 1 total phenolic acids of water cress of table intracellular inhibition of HIV carrying capacity to TZM-B1
4 discuss
TZM-B1 cell deriveds stablize expression HIV-1 infection cell receptor CD4 and accessory receptor in Hela cell lines CXCR4、CCR5.TZM-B1 cells carry the luciferase (Luc) of HIV LTR startups and E.coli β-half as indicator cells Lactoside enzyme (β-gal) gene.Cell starts reporter gene and lives once being infected by HIV or SIV, Tat protein expression transcriptional activations Change expression, you can by the infection conditions of the expression performance virus of luciferase, primary viral and pseudovirus can infect.
This research and utilization TZM-B1 cell tests total phenolic acids of water cress tests adapted strain (SF33) infection cell to inhibition of HIV Inhibiting effect, the results showed that：Total phenolic acids of water cress has good inhibiting effect, two batches experimental result to HIV laboratories adapted strain With repeatability, average IC50For 24.26 μ g/ml.The result shows that total phenolic acids of water cress has certain HIV-resistant activity.
Experimental example 2：Therapeutic effect of the total phenolic acids of water cress to Murine retroviral (Fr.MuLV) infection model
1 materials and methods
1.1 animal BALB/c mouses 48, SPF grades, 17~19g of weight, half male and half female (is purchased from Beijing dimension tonneau China Experimental animal Technology Co., Ltd., animal quality certification number：SCXK (capital) 2012-0001.)
1.2 reagent total phenolic acids of water cress, this research institute extraction (lot number：090303)；Zidovudine tablets, Dongbei Pharmaceutical General Factory Product (lot number：5151102)；Mouse CD4ELISA Kit (lot numbers：201509)、Mouse CD8ELISA Kit (are Shanghai Enzyme-linked bio tech ltd provides)；Trizol (Invitrogen companies, lot number：90803)；Chloroform (Tianjin wind Ship chemical reagent Science and Technology Ltd., lot number：20150226)；Isopropanol (Tianjin East China chemical reagent work, lot number：20140915)； 75% ethyl alcohol (up to commerce and trade Co., Ltd produce, lot number by Wuhan Xinghe：20150101)；One Step SYBY PrimeScriptTM RT-PCR Kit II (Perfect Real Time) kit (TaKaRa companies, lot number：AK2601).
1.3 Fr.MuLV Strain are purchased from American classic culture collecting center (ATCC), are passed in BALB/c mouse body Generation, -70 DEG C of preservations.
1.4 primer m-gapdh-135F:gCTgAgTATgTCgTggAgT；
m-gapdh-135R:gTTCACACCCATCACAAAC；
Friendmurineleukemiavirus-116F:AggATAACCCTCAgAgTCgg；
Friendmurineleukemiavirus-116R:TggACCCAggCAgACTTgTC；
It is synthesized by the prosperous bio tech ltd's design of Beijing AudioCodes.
1.5 electronics assay balances (Ohaus Corp.Brock.WJ.USA)；The A2 type Biohazard Safety Equipments (U.S. Thromo eppendorf)；Centrifuge (German Eppendorf)；Multi-function microplate reader (German PerkinElmer)；It is multi-functional Microplate reader (Thermo companies of the U.S.)；Real-time Thermal Cycler (Thermo companies of the U.S.).
The foundation and administration of 1.6 mouse models will infect the sterile taking-up of spleen of the morbidity mouse of Fr.MuLV, grind, 10% (M/V) suspension is made with DMEM, 4 DEG C, 2000r/min, centrifuges 15min.Take its supernatant abdominal cavity infection mouse, every mouse Inject 0.15ml.After infecting 2h, respectively with zidovudine tablets 90mgkg-1·d-1, total phenolic acids of water cress 500mgkg-1·d-1、 250mg·kg-1·d-1And 125mgkg-1·d-1, according to 0.2ml/10g weight, gastric infusion.Normal group and model pair According to group under equal conditions with distilled water gavage, once a day, continuous 28 days, dissected after weighing within the 29th day, eye socket takes blood, wins Spleen and thymus gland, and weigh.
1.7 Testing index and detection method claim mouse weight before putting to death, and spleen and thymus gland are taken to weigh after execution, with spleen weight The ratio of amount, thymic weight and mouse weight is spleen weight index, thymic weight index；Take parts whole blood observation red blood cell, The variation of leucocyte, hemoglobin；Detach serum, PT-PCR methods[5]Detect spleen virus load；Peripheral blood CD4 +、CD8 +Cell Count simultaneously ratio calculated.
1.8 statistical method experimental datas are handled with 13.0 statistical softwares of SPSS, all data withIt represents, Comparison among groups are examined using variance analysis and t.
2 results
Influence modeling 28 day of 2.1 total phenolic acids of water cress to Fr.MuLV infecting mouses weight, spleen index and thymus index Afterwards, compared with normally group mouse, model group weight is substantially reduced, and spleen index rises appreciably, and thymus index is substantially reduced, and has aobvious Sex differernce (P ＜ 0.01) is write, illustrates model foundation success.Compared with model group, each dosage group weight of total phenolic acids of water cress is It aggravates, spleen index has reduction trend, but statistically there was no significant difference (P ＞ 0.05), total phenolic acids of water cress high dose group Thymus index is increased significantly trend, has otherness (P ＜ 0.05), the results are shown in Table 2.
2 total phenolic acids of water cress of table to mouse weight and the dirty index of device influence (N=8)
Note：Compared with normal group：##P ＜ 0.01；Compared with model group：**P ＜ 0.01,*P ＜ 0.05
Influence of 2.2 total phenolic acids of water cress to Fr.MuLV infecting mouse blood routines is compared with normal group, model group peripheral blood Leucocyte, red blood cell and hemoglobin significantly increase, significant difference (P ＜ 0.01, P<0.05) after, illustrating virus infection Cause the typical erythroleukemia performance of mouse.Compared with model control group, the leucocyte of each dosage group of total phenolic acids of water cress has reduction Trend, but there was no significant difference (P>0.05), total phenolic acids of water cress high dose group red blood cell and hemoglobin are substantially reduced, and are had aobvious Write sex differernce (P<0.05) 3, be the results are shown in Table.
3 total phenolic acids of water cress of table to Fr.MuLV infection blood routine influence (N=8)
Note：Compared with normal group：##P ＜ 0.01；Compared with model group：**P ＜ 0.01,*P ＜ 0.05
2.3 total phenolic acids of water cress are to mouse peripheral blood CD4 +、CD8 +The influence of T cell is compared with normal group, model group CD4 + Content and CD4 +/CD8 +Value is substantially reduced, significant difference (P<0.05) there is immunosupress after, illustrating mouse infection virus. Compared with model group, total phenolic acids of water cress is high, the CD of middle dose group4 +Content and CD4 +/CD8 +Value is apparent to be risen, poor with conspicuousness Different (P ＜ 0.01, P ＜ 0.05), illustrates that total phenolic acids of water cress can effectively improve viral premunition holddown, the results are shown in Table 4。
4 total phenolic acids of water cress of table is to CD4 +、CD8 +T cell percentage and its ratio influence (N=7)
Note：Compared with normal group：##P ＜ 0.01；Compared with model group：**P ＜ 0.01,*P ＜ 0.05
Influence of 2.4 total phenolic acids of water cress to mouse spleen virus load is compared with normal group, in model group mouse spleen Virus load it is significantly raised, there is significant difference (P ＜ 0.05).After administration, total phenolic acids of water cress is high, middle dose group virus carries Amount is decreased obviously, and is had significant difference (P ＜ 0.05), be the results are shown in Table 5.
5 total phenolic acids of water cress of table to Fr.MuLV mouse infection spleen virus loads influence (N=6)
Note：Compared with normal group：#P ＜ 0.05；Compared with model group：*P ＜ 0.05
3 discuss
Fr.MuLV Tobamovirus retrovirus after mouse infection Fr.MuLV viruses, can lead to its weight loss, thymus gland The symptoms such as atrophy, splenomegaly, immunosupress and opportunistic infections.The virus is similar to the symptom of HIV-1 infection people, therefore also has The person model its be mouse AIDS (MAIDS).This research observes total phenolic acids of water cress with Fr.MuLV virus infected mice models While interior resisting virus acts on, its influence to similar HIV-1 infection symptoms is investigated.
This experimental result is shown, after Fr.MuLV virus infected mices, mouse weight declines, and splenomegaly occurs and thymus gland withers The symptom of contracting, CD4 +、CD4 +/CD8 +Value is remarkably decreased, and is occurred a large amount of viruses in spleen, is illustrated modeling achievement.Total phenolic acids of water cress It after administration, has some improvement to mouse splenomegaly, high, middle dose group can significantly improve CD4 +The percentage of T cell improves CD4 +/CD8 +Value, and have apparent inhibiting effect to virus load in mouse spleen.Thus it is speculated that total phenolic acids of water cress can improve machine The immunosuppressive action of body so as to play the role of antivirus action, further needs clinical deep investigation.
Experimental example 3：Total phenolic acids of water cress is in vitro to the inhibiting effect of hiv reverse transcriptase activity
1 experiment material
1.1 drug
Total phenolic acids of water cress, Huang Zhengming seminars independently extract；Nevirapine (NVP), Hai Disainuo pharmacy.
1.2 hiv reverse transcriptase
Hiv reverse transcriptase (is bought in Ambion companies).
1.3 reagent
Dimethyl sulfoxide (DMSO) (DMSO), BIOMOL；Balf serum albumin (BSA), Beijing Sino-U.S. transduction science and technology；DTT, Poly A, PNPP, SIGMA company；DUTP, Shanghai life work；DTTP, oligo (dT)15, the precious biology in Dalian；Tris, Beijing Biodee；Alkali Phosphorus enzyme label streptavidin (APS), Zhong Shan Golden Bridge；MgCl2·6H2O, KCl, NaOH, Beijing Chemical Plant.
1.4 key instrument
96 porocyte culture plates, NUNC companies of Denmark；Low-temperature and high-speed centrifuge, SIGMA 3-18K；Electronic analytical balance, The permanent flat scientific instrument of Shanghai space；Enzyme-linked instrument, U.S. BIO-RAD；Eddy blending machine, its woods Bell's instrument manufacturing of Haimen；Thermostatted water Bath, Grant.
2 experimental methods
2.1 total phenolic acids of water cress are to the inhibiting effect of hiv reverse transcriptase (RT)
Oligo(dT)15It is coated with the preparation of plate
By Oligo (dT)15It is dissolved in the hydrochloride buffer of the 1- methyl-imidazoles of 100mM (pH 7.4), adds in 96 hole enzyme marks In plate, with water-soluble carbodiimide mixing, 4h is reacted in 50 DEG C of water-baths, after reaction with washing lotion (50mmol/L Tris-HCl, pH 7.5) it washes three times, remove unbonded Oligo (dT)15, 96 orifice plates after coating are put into 4 DEG C of preservations.
HIV-1RT Activity determinations
Total phenolic acids of water cress is made into the DMSO solution of 1. 5000 μ g/ml, gradient dilution to 2. 1000 μ g/ml, 3. 500 μ g/ml, 4. 100 μ g/ml respectively take 5 μ l to add in 20 μ l DMSO and 25 μ l PBS.Positive control drug NVP is made into 500 with DMSO and PBS, 100th, 10 μM of liquid is added separately in 96 well culture plates washed with PBS, per 10 μ l of hole, per 3 hole of concentration, is then added in RT Buffer per 70 μ l of hole and the PBS solution containing RT per 20 μ l of hole, make reaction system total volume (wherein contain for 100 μ l 50mmol/L Tris-HCl, pH 8.3,3mmol/L MgCl2, 75mmol/L KCl, 5mmol/L DTT, 0.13mg/ml BSA, 10 μ g/mL poly (A), 0.75 μM of biotin-11-Dutp, 1.5 μM of dTTP and appropriate enzyme), then 37 DEG C of water-bath 1h, With washing lotion (50mmol/L Tris-HCl, pH 7.5,0.15mol/L NaCl, 0.05mmol/L MgCl2, 0.02% Tween20 it) washes three times, removes unbonded free substrate, then 200 μ l 1%BSA are added in per hole, room temperature closing 60min is prevented The non-specific binding of biotin and streptavidin albumen, board-washing, the SA-ALP dilutions (100ng/ml) of 50 μ l of addition per hole, 37 DEG C of water-bath 1h, board-washing add in 100 μ l PNPP (1mg/ml, pH 9.5), 37 DEG C of water-bath 30min per hole；It is added in per 50 μ l of hole The NaOH of 1mol/L terminates reaction, and microplate reader measures A values at 405nm wavelength, to determine the activity of HIV-1RT, while sets not Enzyme negative control, experiment with computing hole A values/negative hole A values (P/N values).
2.2 statistical procedures
Experimental data is with mean ± standard deviationIt represents, with T inspections between group two-by-two compare, and calculate IC50 Value.
3 experimental results
The experimental results showed that total phenolic acids of water cress has apparent inhibiting effect to the activity of hiv reverse transcriptase, with RT control groups (the P that compares that there were significant differences<0.01), and apparent dose-effect relationship is presented in the inhibition of high, medium and low dosage group.It is shown in Table 6.
6 total phenolic acids of water cress of table to hiv reverse transcriptase activity influence (N=3)
Note：Compared with RT control groups, * * P<0.01
4 discuss
HIV is the main pathogens for causing global spread of aids, replicative cycle include the absorption of virus and penetrate, Reverse transcription and integration, the critical stages such as translation process of albumen.Wherein, during hiv reverse transcriptase is HIV genome duplications One of key enzyme inhibits the ideal target spot that hiv reverse transcriptase is also current anti-AIDS drug design.This test water Celery total phenolics are applied in hiv reverse transcriptase, measure its influence to hiv reverse transcriptase activity.The result shows that total phenolic acids of water cress There is apparent inhibitory activity to hiv reverse transcriptase, and have dosage according to lazyness, prompt total phenolic acids of water cress that can pass through inhibition Hiv reverse transcriptase achievees the effect that anti-AIDS.
In conclusion total phenolic acids of water cress experiment in vitro finds that it has stronger HIV-resistant activity to HIV；Experiment in vivo is sent out Existing its can significantly improve CD4 +The percentage of T cell improves CD4 +/CD8 +Value, by improving the immunosuppressive action of body, so as to Play antivirus action；Total phenolic acids of water cress has good inhibiting effect to the activity of hiv reverse transcriptase.In total phenolic acids of water cress body The outer effect for being respectively provided with AntiHIV1 RT activity, function and effect are clear and definite, and are the works by the way that the activity of hiv reverse transcriptase is inhibited to reach AntiHIV1 RT activity With the mechanism of action is more clear and definite, has the potential quality of exploitation Clinical practice.

Claims (12)

1. a kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals, it is characterised in that：With Total phenolic acids of water cress extract bulk pharmaceutical chemicals, bulk pharmaceutical chemicals 55-80% containing total phenolics.
2. the use according to claim 1 that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals On the way, it is characterised in that：Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals, using addition cosolvent for example Tween-80 or Water solubility is made using water-miscible propylene glycol, polyethylene glycol-400 dissolving or using with the methods of organic base forming salt Preparation.
3. according to claim 1 or 2 be used to prepare anti-hiv drug preparation by bulk pharmaceutical chemicals of total phenolic acids of water cress Purposes, it is characterised in that：Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals contains 0.5%-99.5% in component Total phenolic acids of water cress extract bulk pharmaceutical chemicals and 99.5%-0.5% drug excipient.
4. according to claim 1 or 2 be used to prepare anti-hiv drug preparation by bulk pharmaceutical chemicals of total phenolic acids of water cress Purposes, it is characterised in that：Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals, the water containing 1%-60% in component The drug excipient of celery total phenolic acid extract bulk pharmaceutical chemicals and 99%-40%.
5. according to claim 1 or 2 be used to prepare anti-hiv drug preparation by bulk pharmaceutical chemicals of total phenolic acids of water cress Purposes, it is characterised in that：Pharmaceutical preparation containing total phenolic acids of water cress extract bulk pharmaceutical chemicals is oral preparation or parenterai administration agent Type.
6. the use according to claim 5 that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals On the way, it is characterised in that：The oral preparation is selected from tablet, pill, capsule, granule, suspension, dripping pill, oral liquid Body preparation any one of is worked as.
7. the use according to claim 5 that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals On the way, it is characterised in that：The parenteral dosage form is in injection, aerosol, suppository or subcutaneous administration dosage form Any one.
8. the use according to claim 3 that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals On the way, it is characterised in that：The excipient, including one of solvent, disintegrant, corrigent, preservative, colorant and adhesive or Several combinations；The drug of other adapteds is also included in the pharmaceutical excipient.
9. the use according to claim 4 that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals On the way, it is characterised in that：The excipient, including one of solvent, disintegrant, corrigent, preservative, colorant and adhesive or Several combinations；The drug of other adapteds is also included in the pharmaceutical excipient.
10. according to claim 8 or claim 9 be used to prepare anti-hiv drug preparation by bulk pharmaceutical chemicals of total phenolic acids of water cress Purposes, it is characterised in that：The drug of other adapteds included in the pharmaceutical excipient refers to total with the Chinese celery of effective dose Phenolic acid is medicine material, compatibility other allowed the Chinese medicine or chemicals that share.
11. according to claim 1 or 2 be used to prepare anti-hiv drug preparation by bulk pharmaceutical chemicals of total phenolic acids of water cress Purposes, it is characterised in that：The preparation method of total phenolic acids of water cress bulk pharmaceutical chemicals is：
Dry Chinese celery herb is ground into coarse powder, hydrophilic solvent heating and refluxing extraction or the cold soaking diafiltration measured with 10-30 times Extraction, heat carries 2-3 times, 1-3 hours each, and extracting solution is then concentrated under reduced pressure into proportion 1.15- under the conditions of less than 60 DEG C The medicinal extract of 1.30 (70 DEG C)；Primary 4-6 times of hot water of dose is added in, is dissolved by several times, stands 24 hours, is filtered, polyamide on filtrate For the column chromatography of carrier, water is rinsed with water and used：Alcohol ratio is 50：50-5：95 mixed solvent elution, collects water-alcohol elution Liquid is concentrated under reduced pressure under conditions of less than 60 DEG C, is dried in vacuo to obtain total phenolic acids of water cress extract.
12. the use according to claim 11 that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals On the way, it is characterised in that：The preparation method of total phenolic acids of water cress bulk pharmaceutical chemicals is：Dry Chinese celery herb is ground into coarse powder, adds 10 times 70% ethyl alcohol heating and refluxing extraction of amount 2 hours totally 3 times, merges extracting solution, lets cool, filters, then by extracting solution in less than 60 The medicinal extract of proportion 1.15-1.30 (70 DEG C) is concentrated under reduced pressure under the conditions of DEG C；Primary 6 times of hot water of dose are added in, are dissolved by several times, are stood 24 hours, filtering, filtrate was added on 5 times of the crude drug amount that is equivalent to, and on processed good polyamide column, was rinsed with water to efflux in light It after yellow, is eluted with 70% alcohol, collects ethanol eluate until eluent is in faint yellow, depressurized under conditions of less than 60 DEG C Concentration, is dried in vacuo to obtain total phenolic acids of water cress extract.
CN201810237327.9A 2018-03-22 2018-03-22 A kind of purposes that anti-hiv drug preparation is used to prepare using total phenolic acids of water cress as bulk pharmaceutical chemicals Pending CN108186700A (en)

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CN1935167A (en) * 2006-06-02 2007-03-28 黄正明 Water fennel total phenolic acid preparing process, and medicinal preparation using same as active component and use
CN1994341A (en) * 2006-06-02 2007-07-11 黄正明 Preparation process of total phenolic acids of water cress and pharmaceutical formulation using the ingredient as bulk drug and use thereof for treating liver disease

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Publication number Priority date Publication date Assignee Title
CN1935167A (en) * 2006-06-02 2007-03-28 黄正明 Water fennel total phenolic acid preparing process, and medicinal preparation using same as active component and use
CN1994341A (en) * 2006-06-02 2007-07-11 黄正明 Preparation process of total phenolic acids of water cress and pharmaceutical formulation using the ingredient as bulk drug and use thereof for treating liver disease

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