CN106146602A - A kind of preparation method and applications of Cleistanone derivant - Google Patents
A kind of preparation method and applications of Cleistanone derivant Download PDFInfo
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- CN106146602A CN106146602A CN201610559001.9A CN201610559001A CN106146602A CN 106146602 A CN106146602 A CN 106146602A CN 201610559001 A CN201610559001 A CN 201610559001A CN 106146602 A CN106146602 A CN 106146602A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
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Abstract
The present invention relates to organic synthesis and medicinal chemistry art, be specifically related to Cleistanone Cleistanone derivant, preparation method and the purposes in preparation antibacterials thereof.The present invention has synthesized a new Cleistanone Cleistanone derivant, and discloses its preparation method.Pharmacological experiment shows, the Cleistanone Cleistanone derivant of the present invention has antibacterial action, has the value of exploitation antibacterials.
Description
Technical field
The present invention relates to organic synthesis and medicinal chemistry art, be specifically related to Cleistanone Cleistanone derivant, system
Preparation Method and application thereof.
Background technology
The health and lives of the mankind, antibacterials conduct in the diffusion of pathogenic bacterium and the serious threat that strengthens of drug resistance thereof
Routine administration is widely used in acquired immune deficiency syndrome (AIDS), the controlling of organ transplantation and Chronic consumptions (such as cancer, diabetes, uremia etc.)
Treat, although the antimicrobial agent that uses clinically at present (as ketoconazole, amikacin, gentamycin, voriconazole, itraconazole,
Terbinafine, amphotericin, fluconazol etc.) preferable to the curative effect of skin and superficial place infection, but the storage of these antibacterials
Long-pending toxicity is relatively strong, usually causes the stimulation of lesions of liver and kidney, digestive tract, dizziness, allergy etc., so finding the novel of mechanism of action uniqueness
Antibacterials become one of focus of current medicament research and development.
Helicobacter pylori (Helicobacterpylori, Hp) is a kind of Gram-negative spiral bacteria.Research display,
Helicobacter pylori is the primary pathogenic event of acute and chronic gastritis and taste-blindness rate, and may be with gastric cancer and gastric mucosa
The morbidity of associated lymphoid sample tissue (MALT) malignant lymphoma is relevant.Recently, Hp is classified as I class carcinogen by World Health Organization (WHO),
It plays a leading role in stomach cancer development.The scheme that currently a popular treatment Hp infects is to take proton pump inhibitor simultaneously
(PPI) triple therapy of two kinds of antibiotic (clarithromycin, amoxicillin, tetracycline, metronidazole etc. select two kinds) is added.Affect three
The main factor of therapy is considered as the Hp drug resistance to antibacterial；Another serious problems are that proton pump inhibitor can induce and disappears
Changing bad, in a large amount of antibacterial then cause digestive tract, the serious of flora destroys.Therefore, efficient, safe anti-Hp activity one is found
Kind new medicine thing becomes an important and urgent task.
The most global morbidity lungy, in increasing trend, is estimated according to World Health Organization (WHO) (WHO), and the whole world is tied at present
The population that core mycobacteria (Mycobacteriumtuberculosis, MTB) infects accounts for 1/3rd of world population, and wherein 5
～the infected of 10% becomes tuberculosis patient.China occurs active tuberculosis patient 1,300,000 example, wherein infectiousness lung every year
Tuberculosis about 600,000 example, wherein infectiousness pulmonary tuberculosis about 600,000 example, be one of whole world tuberculosis high burden country.
Come out one after another from antituberculotics, make treatment lungy play epoch-making change.Yet with tuberculosis sufferer
The Case management of person the most not ten sectional specification, irregular chemotherapy, abuse antituberculotics, make drug resistance of tuberculosis situation day by day serious,
And the change of drug resistance tends to multi-medicament drug resistance simultaneously, this causes extreme difficulties to preventing and controlling lungy.Therefore
Finding new antituberculotics, the antituberculotics of the most anti-multidrug resistance, to protection people's health, has important
Meaning.
From natural product, find compound or lead compound and carry out structural modification and obtain its derivant, thus obtaining
The potential drug of high-efficiency low-toxicity most has important value.
The compound Cleistanone Cleistanone that the present invention relates to is one to be delivered for 2011
(VanTrinhThiThanhetal.,2011.Cleistanone:
ATriterpenoidfromCleistanthusindochinensiswithaNewCarbonSkeleton.Volume2011,
Issue22, pages4108 4111, August2011) compound, compound Cleistanone Cleistanone is entered by we
Go structural modification, it is thus achieved that a new Cleistanone Cleistanone derivant, and its antibacterial activity has been commented
Valency, it has antibacterial activity.
Summary of the invention
The invention discloses a Cleistanone Cleistanone derivant, its structure is:
Cleistanone Cleistanone derivant (III) of the present invention can be prepared by following method:
(1) Cleistanone Cleistanone (I) reacts the O-bromine obtaining Cleistanone Cleistanone with glycol dibromide
Ethyl derivative (II)；
(2) O-bromoethyl derivant (II) of Cleistanone Cleistanone and dimethylamine generation substitution reaction prepare Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood
Ketone Cleistanone derivant (III).
Further the preparation method of Cleistanone Cleistanone derivant (III) is:
(1) 440mg compound Cleistanone Cleistanone (I) is dissolved in 10mL benzene, in solution, adds four fourths of 0.04g
Base ammonium bromide, the glycol dibromide of 3.760g and 50% sodium hydroxide solution of 6mL；Mixture stirs 24h at 25 degrees Celsius；
After 24h, reactant liquor is poured in frozen water, be extracted twice with dichloromethane immediately, merge organic phase solution；Then to organic facies
Solution washs 3 times with water and saturated aqueous common salt successively, then is dried with anhydrous sodium sulfate, and last concentrating under reduced pressure is removed solvent and produced
Thing crude product；Product crude product silica gel column chromatography is purified, and flowing is mutually: petroleum ether/acetone=100:1, v/v, collects yellow and concentrates
Elution band i.e. obtains the yellow solid of O-bromoethyl derivant (II) of Cleistanone Cleistanone.
(2) the O-bromoethyl derivant of the Cleistanone Cleistanone of 273mg is dissolved in the middle of 20mL acetonitrile, Xiang Qi
The Anhydrous potassium carbonate of middle addition 345mg, the potassium iodide of 84mg and the dimethylamine of 900mg, mixture is heated to reflux 16h；Reaction knot
After bundle, reactant liquor is poured in frozen water, extract three times with equivalent dichloromethane, merge organic facies；Successively with water and saturated aqueous common salt
Organic facies after washing merging, then be dried with anhydrous sodium sulfate, concentrating under reduced pressure is removed solvent and is obtained product crude product；Product crude product
Purifying with silica gel column chromatography, flowing is mutually: petroleum ether/acetone=100:1, v/v, collects faint yellow concentration elution band and is i.e. closed
The faint yellow solid of HUAMU ketone Cleistanone derivant (III).
Compound disclosed by the invention can make pharmaceutically acceptable salt or pharmaceutically acceptable carrier.
Pharmacodynamic experiment shows, Cleistanone Cleistanone derivant (III) of the present invention has preferable antibacterial work
With.The pharmaceutically acceptable salt of the present invention have with its compound as drug effect.
Further, the experiment in vitro of compound III shows, chemicals III has the strongest human body fungi activity, because of
The chemicals III of this present invention is expected to be used for preparing novel anti-human fungi medicine.
Further, the experiment in vitro of compound III shows, compound III has the strongest helicobacter pylori resistant and lives
Property, from the point of view of illustrating for diseases such as acute and chronic gastritis closely-related with helicobacter pylori, duodenal ulcers, chemical combination
Thing III is the compound of a great exploitation potential.It can be directly used for treatment and the preparation of related drugs of corresponding disease.
Further, the experiment in vitro of compound III shows, compound III has the strongest suppression escherichia coli, fluorescence
Pseudomonas, Staphylococcus aureus, Bacillus proteus, the effect of neogenesis cryptococcus, so compound III can be as having antibacterium
The compound of effect, and be expected to be applied in preparing anti-bacterial drug.
Further, according to the result of preliminary test, present invention solid medium dilution method determines this compound pair
Bacillus calmette-guerin vaccine, mycobacterium tuberculosis type strain H37Rv strain and the minimum of substance of medicines-resistant branched tubercle bacillus (MDRMTB) three kinds of tulases
Mlc, experimental result confirms that compound III has the strongest anti-tubercle bacillus and anti-drug resistance tulase activity, can be as controlling
Treat the lead compound of tubercle bacillus affection disease it can also be used to tubercular drugs is treated in preparation.
The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not by concrete real
Execute any restriction of example, but be defined in the claims.Detailed description of the invention
The preparation of embodiment 1 compound Cleistanone Cleistanone
The document that the preparation method of compound Cleistanone (I) is delivered with reference to VanTrinhThiThanh et al.
(VanTrinhThiThanhetal.,2011.Cleistanone:ATriterpenoidfromCleistanthusindochin
EnsiswithaNewCarbonSkeleton.Volume2011, Issue22, pages4108 4111, August2011) side
Method.
The synthesis of O-bromoethyl derivant (II) of embodiment 2 Cleistanone Cleistanone
Compound I (440mg, 1.00mmol) is dissolved in 10mL benzene, in solution, adds tetrabutyl ammonium bromide (TBAB)
(0.04g), glycol dibromide (3.760g, 20.00mmol) and 50% sodium hydroxide solution of 6mL.Mixture is Celsius 25
Degree stirring 24h.After 24h, reactant liquor is poured in frozen water, be extracted twice with dichloromethane immediately, merge organic phase solution.So
Washing organic phase solution 3 times with water and saturated aqueous common salt successively afterwards, then be dried with anhydrous sodium sulfate, last concentrating under reduced pressure is removed
Solvent obtains product crude product.Product crude product silica gel column chromatography purification (flowing is mutually: petroleum ether/acetone=100:1, v/v), receives
Collection yellow concentrates elution band i.e. to obtain the yellow solid (344mg, 63%) of compound II.
1HNMR (500MHz, DMSO-d6) δ 5.04 (s, 1H), 4.82 (s, 1H), 3.94 (d, J=26.5Hz, 1H), 3.87
(d, J=26.5Hz, 2H), 3.57 (s, 2H), 2.40 (d, J=14.0Hz, 1H), 2.39 (d, J=14.0Hz, 1H), 2.27 (s,
1H), 2.21 (s, 1H), 2.15 (s, 1H), 1.82 (s, 1H), 1.62 (s, 2H), 1.57 (d, J=3.3Hz, 1H), 1.54 (d, J
=3.3Hz, 1H), 1.50 (d, J=1.2Hz, 1H), 1.47 (d, J=1.2Hz, 1H), 1.39 (d, J=15.3Hz, 2H), 1.34
(d, J=15.3Hz, 1H), 1.26 (dd, J=32.6,13.7Hz, 4H), 1.13 (d, J=18.0Hz, 2H), 1.05 (s, 6H),
0.98(s,1H),0.88(s,12H),0.78(s,3H),0.74(s,1H)。
13CNMR(125MHz,DMSO-d6)δ216.59(s),154.50(s),105.23(s),74.63(s),69.85
(s),59.71(s),52.55(s),51.21(s),47.92(s),44.10(s),42.25(s),41.73(s),40.64(s),
40.16 (s), 38.88 (s), 38.65 (s), 37.21 (s), 36.23 (s), 33.34 (d, J=1.1Hz), 32.96 (s), 29.91
(s),27.18(s),26.03(s),24.23(s),23.96(s),20.77(s),18.48(s),17.98(s),16.93(s)。
HRMS(ESI)m/z[M+H]+calcdforC32H52BrO2:547.3151；found547.3159.
The synthesis of O-(dimethylamino) ethyl derivative (III) of embodiment 3 Cleistanone Cleistanone
Compound II (273mg, 0.5mmol) is dissolved in the middle of 20mL acetonitrile, be added thereto to Anhydrous potassium carbonate (345mg,
2.5mmol), potassium iodide (84mg, 0.5mmol) and dimethylamine (900mg, 20mmol), mixture is heated to reflux 16h.Reaction knot
After bundle, reactant liquor is poured in frozen water, extract three times with equivalent dichloromethane, merge organic facies.Successively with water and saturated aqueous common salt
Organic facies after washing merging, then be dried with anhydrous sodium sulfate, concentrating under reduced pressure is removed solvent and is obtained product crude product.Product crude product
Purify (flowing is mutually: petroleum ether/acetone=100:1, v/v) with silica gel column chromatography, collect faint yellow concentration elution band and i.e. obtain
The faint yellow solid (160.7mg, 61%) of compound III.
1HNMR(500MHz,DMSO-d6)δ5.02(s,1H),4.81(s,1H),3.85(s,1H),3.51(s,2H),
2.84 (s, 6H), 2.64 (s, 2H), 2.38 (d, J=13.0Hz, 2H), 2.26 (s, 1H), 2.17 (s, 1H), 2.17 (s, 1H),
1.83 (s, 1H), 1.66 (s, 2H), 1.54 (d, J=6.0Hz, 2H), 1.46 (d, J=0.9Hz, 2H), 1.34 (d, J=
13.5Hz, 3H), 1.26 (dd, J=31.1,13.9Hz, 4H), 1.16 (d, J=13.5Hz, 2H), 1.07 (s, 6H), 0.98 (s,
1H),0.85(s,12H),0.76(s,3H),0.73(s,1H).
13CNMR(125MHz,DMSO-d6)δ216.60(s),154.51(s),105.19(s),74.62(s),65.68(s),
59.75(s),57.63(s),52.57(s),51.18(s),47.87(s),45.54(s),44.12(s),42.29(s),41.79
(s),40.62(s),40.12(s),38.82(s),38.67(s),37.25(s),36.29(s),33.32(s),32.92(s),
29.85(s),27.20(s),26.07(s),24.29(s),23.94(s),20.73(s),18.42(s),18.00(s),16.97
(s).
HRMS(ESI):m/z[M+H]+calcdforC34H58NO2:512.4468；found:512.4463.
Embodiment 4 compound involved in the present invention II and the preparation of III tablet
Taking the one in the middle of 20 g of compound II or III or its pharmaceutically acceptable salt, the routine of tablet is prepared in addition
Adjuvant 180 grams, mixing, conventional tablet presses makes 1000.
Embodiment 5 compound involved in the present invention II and the preparation of III capsule:
Taking the one in the middle of 20 g of compound II or III or its pharmaceutically acceptable salt, the normal of capsule is prepared in addition
Rule adjuvant such as starch 180 grams, mixes, encapsulated makes 1000.
O-(dimethylamino) ethyl derivative (III) the human body fungi activity of embodiment 6 Cleistanone Cleistanone
The experiment of human body fungi activity is the method using concentration dilution, measures in triplicate every time, and the pathogen of test has red
Color trichophyton, microsporum canis and trichophyton tonsurans, bacterial concentration is 105/mL.The O-of Cleistanone Cleistanone
(dimethylamino) ethyl derivative (III) initial concentration is 50.00 μ g/mL (5% dimethyl sulfoxide DMSO), and gradient dilution is extremely
(ultimate density of formation has: 25.00, in 96 orifice plates for 0.10 μ g/mL, the bacterium solution of equivalent volumes and test sample mixed culture
12.50,6.25,3.13,1.56,0.78,0.39,0.20,0.10 and 0.05 μ g/mL), Human Fungi cultivation temperature is respectively 28
DEG C, observe after incubation time 24h, if finding, in the disposal hole not having bacterium colony to be formed, that minimum concentration is that sample is minimum antibacterial
Concentration, i.e. MIC value.This experiment positive control is ketoconazole, and chemicals III human body fungus the results are shown in Table 1.
Table 1 chemicals III human body fungus MIC value (μ g/mL)
Conclusion: chemicals III has the strongest human body fungi activity, and therefore the chemicals III of the present invention is expected to be used for system
Standby novel anti-human fungi medicine.
Embodiment 7 compound III anti-helicobactor pylori activity
1) strains tested
Helicobacter pylori (Helicobacterpylori, Hp) reference culture ATCC43504 is purchased from U.S.'s Culture Collection
(AmericanTypeCultureCollection, ATCC).13 strain Hp clinical strains are picked up from Nanjing drum tower hospital Dndoscope Laboratory and are connect
By gastroscopic patient；Patient to peptic ulcer, duodenal bulbar inflammation or gastritis verrucosa in continuous gastroscopy, first
It is defined as Hp positive through RUT experiment, then takes antral gastric mucosa 1-2 block, be inoculated in after chopping containing 8% horse serum, front three
Oxygen benzyl Aminometradine (trimethropin, TMP) 1.25g/L, polymyxin (polymyxin) B2500U/L, vancomycin
(vancomycin) in the Columbia selectivity agar culture medium of 10mg/L, in 37 DEG C under micro-oxygen environment (5%O2,10%
CO2 and 85%N2) cultivate 72 hours.Collecting antibacterial, through smear Gram’s staining, oxidase, catalase and urease are accredited as sun
After property, passing on pure culture, obtained strains is as experimental strain.
2) strain culturing
We use micro-aerobic bag (purchased from Shanghai Medical Univ) to carry out the strain culturing of HP, and it can produce Hp by chemical reaction
Required micro-aerobic environment.
3) biological activity determination
Use paper disk method (Microbiologicalpapermethod) to measure the compound inhibitory action to helicobacter pylori, use
Agar dilution measures the minimum inhibitory concentration (minimalinhibitoryconcentration, MIC) of test sample.
I. paper disk method experiment
(A) prepare culture medium by the Columbia culture medium for preparing after high pressure steam sterilization, be cooled to 50-60 DEG C, add 8%
Horse serum or Sheep Blood, mixing is poured in the most sterilized culture dish, and every ware 7-10ml, culture medium thickness is that 1.5mm is (aseptic
Operation).
(B) switching experimental bacteria (being coated with bacterium) takes the 108CFU/ml (1OD660=108CFU/ml) diluted with microscale sampler
The bacteria suspension 0.1ml of Hp spreads upon suitable culture dish surface equably.It is inverted in 37 DEG C of drying bakers and takes out after 15min, mesh
The agar surface that makes be dried, standby.
(C) patch sample scraps of paper microscale sampler take 6 μ l testing sample (mass concentration 2mg/ml) inject the most sterilized
On circle filter paper.With the blank scraps of paper of the aseptic nipper tweezer scraps of paper containing sample and comparison, it is close to by sterile working's scraps of paper respectively
Containing bacterio-agar surface, patch a piece of paper sheet at a certain distance.Every kind of bacterium is cooked 3 wares, and acquired results seeks its meansigma methods.
(D) each plate is placed in micro-aerobic bag by cultivation, and sealing is opened gas generator, then is placed in 37 DEG C of incubators
Cultivate 72h.
(E), after surveying antibacterial circle diameter taking-up flat board, the size of antibacterial circle diameter around each scraps of paper is measured respectively.With reference to comparison
The result of group, can draw the result of testing sample sensitive experiment.In triplicate.
II. agar dilution measures MIC
(A) first compound dimethyl sulfoxide (DMSO) solution of test is configured to 0.5mg/ml's by the preparation of Drug plates
Mother solution, then dilute with sterilized water, finally it is made into 100.0,80.0,60.0,45.0,40.0,35.0,30.0,25.0,20.0,
The concentration series of 15.0,10.0,5.0 and 2.5 μ g/ml, DMSO concentration in media as well is less than 1%.The test that 1ml is prepared
Compound solution be incubated the 8ml Columbia medium in 50 DEG C, separately add the horse serum that 1ml is incubated in 50 DEG C and fully mix,
Cast in culture dish cooling (in culture dish final concentration of the 10.0 of chemicals, 8.0,6.0,4.5,4.0,3.5,3.0,
2.5,2.0,1.5,1.0,0.5 and 0.25 μ g/ml, if there being bacteriostasis, then during i.e. these are worthwhile one of MIC value).
(B) switching experimental bacteria (being coated with bacterium) draws the bacteria suspension of the 1 × 108CFU/mlHp diluted with microscale sampler
0.1ml spreads upon culture dish surface equably, is inverted in 37 DEG C of drying bakers and takes out after 15min, and purpose makes agar surface be dried,
Standby.
(C) determine MIC by test dish in micro-aerobic bag (containing: 85%N2,10%CO2 and 5%O2), be incubated 37 DEG C
: g cultivates 72 hours, observes Hp growing state, contrasts with blank group, minimum with the sample entirely without bacteria growing
Concentration is minimum inhibitory concentration (minimalinhibitoryconcentration, MIC) value.Positive control is ampicillin
(Ampicillin)。
Experimental result
Experiment in vitro shows, chemicals III has the strongest anti Helicobacter pylori activity, and paper disk method shows its antibacterial circle diameter
For 18mm (ATCC43504).Show that it can completely inhibit 5 random clinical strains and 1 reference culture with agar dilution
(ATCC43504) growth, minimal inhibitory concentration (MIC) is 2.5 μ g/ml.Making positive control with ampicillin, it is to 6 strains
The Cmin (MIC) that test bacterium completely inhibits is 1.5 μ g/ml.
Conclusion: chemicals III has stronger suppression helicobacter pylori activity, illustrate for H. pylori nectar
From the point of view of the diseases such as the acute and chronic gastritis of cut pass, duodenal ulcer, chemicals III is the change of a great exploitation potential
Compound.It can be directly used for treatment and the preparation of related drugs of corresponding disease.
Embodiment 8 chemicals III antibacterial activity
Antibacterial activity test is the method using concentration dilution, measures in triplicate every time, and test pathogen has escherichia coli, glimmering
Light pseudomonas, Staphylococcus aureus, Bacillus proteus, neogenesis cryptococcus, bacterial concentration is 105/mL.Compound III initiates
Concentration is 50.00 μ g/mL (5% dimethyl sulfoxide DMSO), and gradient dilution is to 0.10 μ g/mL, the bacterium solution of equivalent volumes and test
Sample mix cultivate in 96 orifice plates (ultimate density of formation has: 25.00,12.50,6.25,3.13,1.56,0.78,0.39,
0.20,0.10 and 0.05 μ g/mL), antibacterial culturing temperature is respectively 37 DEG C, observes after incubation time 24h, if finding do not have bacterium colony
In the disposal hole formed, that minimum concentration is sample lowest concentration of antimicrobial, i.e. MIC value.Compound III the anti-bacterial result is shown in Table
2。
Table 2 compound III antibacterium MIC value (μ g/mL)
Conclusion: compound III has the strongest antibacterial activity, and therefore the compound III of the present invention is expected to be used for preparation newly
Type anti-bacterial drug.
Embodiment 9 compound III anti-tubercle bacillus activity
(1) solid medium dilution method measures compound III anti-bacillus calmette-guerin vaccine (BCG) absolute concentration
From inclined-plane, scrape BCG cultures, join in 3mlMiddlebrook7H9 broth bouillon, add a small amount of glass
Pearl, screws test tube cap, and in vortex oscillator, high vibration grinds, and compares with standard Maxwell opacity tube (MacFarlandNo.1)
Turbid, i.e. it is made into bacillus calmette-guerin vaccine (BCG) bacteria suspension of 1mg/ml.
Compound III DMSO is made into the stock solution of high concentration, dilutes stock solution with the aseptic ultra-pure water of tween 80 containing 5%
To desired concn, by required dosage, the compound III diluted is joined 4mlMiddlebrook7H11 agar culture medium (should
Culture medium 121 DEG C of high pressure steam sterilizations 15 minutes, it is cooled to 50～55 DEG C), mixing, to make containing compound III, concentration is divided
Not Wei 6.0ug/ml, 4.0ug/ml, 3.0ug/ml, 2.0ug/ml, 1.5ug/ml, 1.0ug/ml, 0.75ug/ml, 0.5ug/ml
Isocyatic slant medium.
Bacillus calmette-guerin vaccine (BCG) the bacteria suspension inoculating loop that concentration is 1mg/ml is dipped ring of numbers, is inoculated in containing compound respectively
In the culture medium of III series concentration and blank medium slant, it is placed in 37 DEG C and cultivates 4～8 weeks, observation experiment result, knot
Fruit is as shown in table 3.
In the present embodiment, Middlebrook7H9 broth bouillon used and Middlebrook7H11 agar culture medium are this
Skilled person carries out conventional culture medium when tulase is cultivated, and its formula uses conventional formulation.
(2) solid medium dilution method measures compound III Killing Mycobacterium Tuberculosis type strain H37Rv strain absolute concentration
From inclined-plane, scrape mycobacterium tuberculosis type strain H37Rv strain culture, join 3mlMiddlebrook7H9 broth cultivation
Supporting in base, add a small amount of bead, screw test tube cap, in vortex oscillator, high vibration grinds, with standard Maxwell opacity tube
(MacFarlandNo.1) ratio is turbid, is i.e. made into the H37Rv strain bacteria suspension of 1mg/ml.
Compound III is made into DMSO respectively the stock solution of high concentration, dilutes with the aseptic ultra-pure water of tween 80 containing 5%
The compound III diluted, to desired concn, is joined 4mlMiddlebrook7H11 agar by required dosage and cultivates by stock solution
Base (this culture medium 121 DEG C of high pressure steam sterilizations 15 minutes, be cooled to 50～55 DEG C), mixing, make containing compound III,
Concentration is respectively 6.0ug/ml, 4.0ug/ml, 3.0ug/ml, 2.0ug/ml, 1.5ug/ml, 1.0ug/ml, 0.75ug/ml,
The isocyatic slant medium of 0.5ug/ml.
The H37Rv strain bacteria suspension inoculating loop that concentration is 1mg/ml is dipped ring of numbers, is inoculated in respectively containing compound III system
In the culture medium of row concentration and blank medium slant, it is placed in 37 DEG C and cultivates 4～8 weeks, observation experiment result, result such as table
Shown in 3.
(3) solid medium dilution method measures compound III Ad tuberculosis to be clinically separated resistance to ISREMTB strain absolute
Concentration
From inclined-plane, scrape mycobacterium tuberculosis be clinically separated resistance to ISREMTB strain (resistance to isoniazid, streptomycin, rifampicin, ethamine fourth
Alcohol mycobacterium tuberculosis is clinically separated post) culture, join in 3mlMiddlebrook7H9 broth bouillon, add a small amount of
Bead, screws test tube cap, and in vortex oscillator, high vibration grinds, with standard Maxwell opacity tube (MacFarlandNo.1)
Ratio is turbid, is i.e. made into the bacteria suspension of 1mg/ml.
Compound III is made into DMSO respectively the stock solution of high concentration, dilutes with the aseptic ultra-pure water of tween 80 containing 5%
The compound III diluted, to desired concn, is joined 4mlMiddlebrook7H11 agar by required dosage and cultivates by stock solution
Base (this culture medium 121 DEG C of high pressure steam sterilizations 15 minutes, be cooled to 50～55 DEG C), mixing, make containing compound III,
Concentration is respectively 6.0ug/ml, 4.0ug/ml, 3.0ug/ml, 2.0ug/ml, 1.5ug/ml, 1.0ug/ml, 0.75ug/ml,
The isocyatic slant medium of 0.5ug/ml.
The mycobacterium tuberculosis that concentration is 1mg/ml is clinically separated resistance to ISREMTB strain bacteria suspension inoculating loop and dips number
Ring, is inoculated in the culture medium containing compound III series concentration and blank medium slant respectively, is placed in 37 DEG C and cultivates 4
～8 weeks, observation experiment result, result is as shown in table 3.
Table 3 solid medium dilution method measures compound III anti-tubercle bacillus absolute concentration result
Conclusion: compound III has the strongest anti-tubercle bacillus and anti-drug resistance tulase activity, can be as treatment tubercle bacillus affection
The lead compound of disease is it can also be used to tubercular drugs is treated in preparation.
Claims (2)
1. a preparation method for Cleistanone derivant, comprises the following steps:
(1) Cleistanone (I) reacts O-bromoethyl derivant (II) obtaining Cleistanone with glycol dibromide；
(2) O-bromoethyl derivant (II) of Cleistanone and dimethylamine generation substitution reaction prepare Cleistanone derivant
(III)。
2. the Cleistanone derivant described in claim 1 is applied to treat the acute gastritis, chronic that helicobacter pylori causes
Gastritis, gastric ulcer and duodenal ulcer.
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| CN201610559001.9A CN106146602A (en) | 2014-06-25 | 2014-06-25 | A kind of preparation method and applications of Cleistanone derivant |
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| CN109777460A (en) * | 2019-01-30 | 2019-05-21 | 虞定生 | A kind of acicular petroleum coke and its processing technology |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104447938B (en) * | 2014-11-05 | 2016-11-30 | 南京大学 | O-(piperazinyl) ethyl derivative of Cleistanone, preparation method and its usage |
| CN105193816A (en) * | 2015-07-23 | 2015-12-30 | 南京赋海澳赛医药科技有限公司 | Composition and application thereof in antibacterial drugs |
| CN105078989A (en) * | 2015-09-08 | 2015-11-25 | 苏州贺澳德生物医药科技有限公司 | Composition 77083001030592 and application thereof in antibacterial drugs |
| CN105616423A (en) * | 2015-12-29 | 2016-06-01 | 南京大学 | Composition and application of composition to antibacterial medicine |
| CN106718391A (en) * | 2016-11-29 | 2017-05-31 | 扬州大学（如皋）花木产业研究院 | A kind of preparation method of flower-tree saving solution |
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| CN104086617B (en) * | 2014-06-25 | 2016-05-18 | 吉林大学 | Close dimethylamine derivative, the preparation method and its usage of flowers and trees ketone Cleistanone |
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| CN103446114A (en) * | 2013-09-18 | 2013-12-18 | 南京广康协生物医药技术有限公司 | Applications of Sarcaboside A in antibacterial medicament |
| CN103462989A (en) * | 2013-09-18 | 2013-12-25 | 南京广康协生物医药技术有限公司 | Application of Lycojaponicumin B in preparation of medicines for resisting human body fungi |
| CN103446115A (en) * | 2013-09-18 | 2013-12-18 | 南京广康协生物医药技术有限公司 | Applications of Sarcaboside A in medicament against human fungus |
| CN103462968B (en) * | 2013-09-22 | 2015-11-25 | 刘岩 | Incarviatone A is preparing the application in anti-bacterial drug |
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| CN104086617B (en) * | 2014-06-25 | 2016-05-18 | 吉林大学 | Close dimethylamine derivative, the preparation method and its usage of flowers and trees ketone Cleistanone |
Non-Patent Citations (2)
| Title |
|---|
| QINGGANG JI ET AL.: "Benzothieno[3,2-b]indole derivatives as potent selective estrogen receptor modulators", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》 * |
| VAN TRINH THI THANH ET AL.: "Cleistanone: A Triterpenoid from Cleistanthus indochinensis with a New Carbon Skeleton", 《EUROPEAN JOURNAL ORGANIC CHEMISTRY》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109777460A (en) * | 2019-01-30 | 2019-05-21 | 虞定生 | A kind of acicular petroleum coke and its processing technology |
| CN109777460B (en) * | 2019-01-30 | 2021-07-16 | 虞定生 | Acicular petroleum coke and processing technology thereof |
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| Publication number | Publication date |
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| CN104083377A (en) | 2014-10-08 |
| CN106188212A (en) | 2016-12-07 |
| CN104083377B (en) | 2016-12-28 |
| CN106146603A (en) | 2016-11-23 |
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