CN104188984A - Application of Cleistanone O-(morpholinyl)ethyl derivative in preparation of antibacterial drugs - Google Patents
Application of Cleistanone O-(morpholinyl)ethyl derivative in preparation of antibacterial drugs Download PDFInfo
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Abstract
The invention relates to the field of organic synthesis and pharmaceutical chemistry and particularly relates to a Cleistanone O-(morpholinyl)ethyl derivative, a preparation method thereof and application thereof in the preparation of antibacterial drugs. According to the invention, a novel Cleistanone O-(morpholinyl)ethyl derivative is synthesized, and a preparation method of the novel Cleistanone O-(morpholinyl)ethyl derivative is disclosed. Shown by pharmacological experiments, the Cleistanone O-(morpholinyl)ethyl derivative disclosed by the invention has an antibacterial effect and has a value in the development of the antibacterial drugs.
Description
Technical field
The present invention relates to organic synthesis and pharmaceutical chemistry field, be specifically related to O-(morpholinyl) ethyl derivative, the preparation method and its usage of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone.
Background technology
The mankind's health and lives in the enhancing serious threat of the diffusion of pathogenic bacterium and drug resistance thereof, antibacterials are widely used in acquired immune deficiency syndrome (AIDS) as routine administration, organ transplantation and Chronic consumptions are (as cancer, diabetes, uremia etc.) treatment, although the antimicrobial agent of using clinically is at present (as ketoconazole, amikacin, gentamycin, voriconazole, itraconazole, terbinafine, amphotericin, fluconazol etc.) better to the curative effect of skin and superficial place infection, but the cumulative toxicity of these antibacterials is stronger, usually cause lesions of liver and kidney, digestive tract stimulates, dizzy, irritated etc., so find the novel antibacterial medicine of mechanism of action uniqueness, become one of focus of current medicament research and development.
Helicobacter pylori (Helicobacter pylori, Hp) is a kind of Gram-negative spiral bacteria.Studies show that, helicobacter pylori is the main pathogenesis of acute and chronic gastritis and Stomach duodenum ulcer, and may be relevant with the morbidity of gastric cancer stomach function regulating mucosa-associated lymphoid tissue (MALT) malignant lymphoma.Recently, World Health Organization (WHO) is classified as I class carcinogen by Hp, and it plays a leading role in gastric cancer development.The scheme that at present popular treatment Hp infects is to take the triple therapy that proton pump inhibitor (PPI) adds two kinds of antibiotic (clarithromycin, amoxicillin, tetracycline, metronidazole etc. select two kinds) simultaneously.The main factor that affects triple therapy is considered to the drug resistance of Hp to antibacterial; Another serious problems are that proton pump inhibitor can bring out dyspepsia, and a large amount of antibacterial cause the serious destruction of flora in digestive tract.Therefore, find the active kind new medicine thing of efficient, safe anti-Hp and become an important and urgent task.
Global morbidity lungy is in recent years increases trend, according to World Health Organization (WHO) (WHO), estimate, the whole world is subject to mycobacterium tuberculosis (Mycobacterium tuberculosis at present, MTB) population infecting accounts for 1/3rd of world population, and wherein the infected of 5～10% becomes tuberculosis patient.There are every year active tuberculosis patient 1,300,000 examples in China, infectiousness pulmonary tuberculosis approximately 600,000 examples wherein, and wherein infectiousness pulmonary tuberculosis approximately 600,000 examples, are one of the high burden of global tuberculosis countries.
From antituberculotics, come out one after another, make treatment lungy play epoch-making variation.Yet due to the treatment management of tuberculosis patient standard very not still, irregular chemotherapy, abuse antituberculotics, makes drug resistance of tuberculosis situation day by day serious, and the variation of drug resistance more trends towards multi-medicament drug resistance simultaneously, and this causes very big difficulty to preventing and controlling lungy.Therefore find new antituberculotics, the antituberculotics of especially anti-multidrug resistance is to protection people's health, significant.
From natural product, find compound or lead compound and carry out structural modification and obtain its derivant, thereby the potential drug that obtains high-efficiency low-toxicity has important value.
At present for antibacterial treatment, there is no clinically specific medicament, most of medicine has inevitable toxic and side effects when alleviating antibacterial symptom, from natural product, find compound or lead compound and carry out structural modification and obtain its derivant, thereby the potential drug that obtains high-efficiency low-toxicity has important value.
The compound Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone the present invention relates to is one and within 2011, delivers (Van Trinh Thi Thanh et al., 2011.Cleistanone:A Triterpenoid from Cleistanthus indochinensis with a New Carbon Skeleton.
volume2011, Issue22,pages4108 – 4111, August2011) compound, we have carried out structural modification to compound Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, obtained O-(morpholinyl) ethyl derivative of a new Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, and its antibacterial activity is evaluated, it has antibacterial activity.
Summary of the invention
O-(morpholinyl) ethyl derivative that the invention discloses a Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, its structure is:
O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr of the present invention wood ketone Cleistanone can be prepared by method below:
(1) Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone (I) reacts the O-bromoethyl derivant (II) that obtains Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone with glycol dibromide;
(2) the O-bromoethyl derivant (II) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone and morpholine generation substitution reaction make O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone.
Further the preparation method of O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is:
(1) 440mg compound Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone (I) is dissolved in to 10mL benzene, to the tetrabutyl ammonium bromide that adds 0.04g in solution, 50% sodium hydroxide solution of the glycol dibromide of 3.760g and 6mL; Mixture stirs 24h at 25 degrees Celsius; After 24h, reactant liquor is poured in frozen water, used immediately dichloromethane extraction twice, merge organic phase solution; Then to organic phase solution successively water and saturated common salt water washing 3 times, then use anhydrous sodium sulfate drying, last concentrating under reduced pressure is removed solvent and is obtained product crude product; Product crude product purification by silica gel column chromatography, mobile phase is: petroleum ether/acetone=100:1, v/v, collects the yellow yellow solid of concentrating elution band to obtain the O-bromoethyl derivant (II) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone.
(2) the O-bromoethyl derivant (II) of the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of 273mg is dissolved in the middle of 20mL acetonitrile, adds wherein the Anhydrous potassium carbonate of 345mg, the potassium iodide of 84mg and the morpholine of 1742mg, mixture reflux 8h; After reaction finishes, reactant liquor is poured in 30mL frozen water, used equivalent dichloromethane extraction three times, merge organic facies; Water and saturated common salt water washing merge organic facies afterwards successively, then use anhydrous sodium sulfate drying, and concentrating under reduced pressure is removed solvent and obtained product crude product; Product crude product purification by silica gel column chromatography, mobile phase is: petroleum ether/acetone=100:0.5, v/v, collects the yellow yellow solid 185.3mg that concentrates elution band to obtain O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone.
Compound disclosed by the invention can be made pharmaceutically acceptable salt or pharmaceutically acceptable carrier.
Pharmacodynamic experiment shows, O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of the present invention has good antibacterial action.Pharmaceutically acceptable salt of the present invention and its compound have same drug effect.
Further, the experiment in vitro of O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone shows, chemicals III has very strong anti-human body fungi activity, and O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone therefore of the present invention is expected to be used to prepare novel anti-human body fungi-medicine.
Further, the experiment in vitro of O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone shows, O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has very strong anti Helicobacter pylori activity, explanation for the disease such as the closely-related acute and chronic gastritis of helicobacter pylori, duodenal ulcer, O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is the compound of a great exploitation potential for its.It can be directly used in the treatment of corresponding disease and the preparation of related drugs.
Further, the experiment in vitro of O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone shows, O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has the effect of very strong inhibition escherichia coli, fluorescent pseudomonas, Staphylococcus aureus, Bacillus proteus, neogenesis cryptococcus, so O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone can be used as the compound with antibacterial actions, and is expected to be applied in preparing anti-bacterial drug.
Further, according to the result of preliminary test, the minimal inhibitory concentration of this compound to bacillus calmette-guerin vaccine, the H37Rv strain of mycobacterium tuberculosis type strain and three kinds of tulases of substance of medicines-resistant branched tubercle bacillus (MDR MTB) that the present invention has used solid medium By Dilution, experimental result confirms that O-(morpholinyl) ethyl derivative of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has very strong anti-tubercle bacillus and anti-drug resistance tulase is active, can be used as the lead compound for the treatment of tubercle bacillus affection disease, also can be used for preparation treatment tuberculosis medicine.
The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not subject to any restriction of specific embodiment, but be limited by claim.
The specific embodiment
The preparation of embodiment 1 compound Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Document (the Van Trinh Thi Thanh et al. that the preparation method of compound Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone (I) is delivered with reference to people such as Van Trinh Thi Thanh, 2011.Cleistanone:A Triterpenoid from Cleistanthus indochinensis with a New Carbon Skeleton.Volume2011, Issue22, pages4108 – 4111, method August2011).
Synthesizing of the O-bromoethyl derivant (II) of embodiment 2 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Compound I (440mg, 1.00mmol) is dissolved in to 10mL benzene, in solution, adds tetrabutyl ammonium bromide (TBAB) (0.04g), 50% sodium hydroxide solution of glycol dibromide (3.760g, 20.00mmol) and 6mL.Mixture stirs 24h at 25 degrees Celsius.After 24h, reactant liquor is poured in frozen water, used immediately dichloromethane extraction twice, merge organic phase solution.Then to organic phase solution successively water and saturated common salt water washing 3 times, then use anhydrous sodium sulfate drying, last concentrating under reduced pressure is removed solvent and is obtained product crude product.(mobile phase is product purification by silica gel column chromatography for crude product: petroleum ether/acetone=100:1, v/v), collect the yellow yellow solid (344mg, 63%) of concentrating elution band to obtain Compound I I.
1H?NMR(500MHz,DMSO-d
6)δ5.04(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)。
13C?NMR(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]
+calcd?for?C
32H
52BrO
2:547.3151；found547.3159.
Synthesizing of O-(morpholinyl) ethyl derivative (III) of embodiment 3 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Compound I I (273mg, 0.5mmol) is dissolved in the middle of 20mL acetonitrile, adds wherein Anhydrous potassium carbonate (345mg, 2.5mmol), potassium iodide (84mg, 0.5mmol) and morpholine (1742mg, 20mmol), mixture reflux 8h.After reaction finishes, reactant liquor is poured in 30mL frozen water, used equivalent dichloromethane extraction three times, merge organic facies.Water and saturated common salt water washing merge organic facies afterwards successively, then use anhydrous sodium sulfate drying, and concentrating under reduced pressure is removed solvent and obtained product crude product.(mobile phase is product purification by silica gel column chromatography for crude product: petroleum ether/acetone=100:0.5, v/v), collect the yellow yellow solid (185.3mg, 67%) of concentrating elution band to obtain O-(morpholinyl) ethyl derivative of Cleistanone.
1H?NMR(500MHz,DMSO-d6)δ5.11(s,1H),4.81(s,1H),4.39(s,1H),3.51(d,J＝17.7Hz,6H),2.59(s,2H),2.47(s,4H),2.32(t,J＝36.5Hz,2H),2.24(s,1H),2.20(s,1H),2.11(s,1H),1.79(d,J＝90.0Hz,3H),1.59(d,J＝7.0Hz,2H),1.40(d,J＝4.2Hz,2H),1.35(d,J＝14.5Hz,3H),1.30–1.16(m,4H),1.11(d,J＝6.5Hz,2H),1.02(s,6H),0.82(d,J＝8.5Hz,13H),0.73(s,3H),0.65(s,1H).
13C?NMR(125MHz,DMSO-d6)δ216.37(s),154.55(s),105.98(s),73.28(s),69.13(s),66.17(s),58.23(s),54.02(s),53.01(s),52.77(s),51.83(s),48.12(s),46.23(s),42.97(s),41.13(s),40.99(s),39.93(s),38.92(s),38.67(s),37.13(s),36.59(s),33.21(s),32.13(s),29.41(s),27.38(s),26.01(s),24.33(s),23.12(s),20.13(s),19.13(s),17.25(s),16.16(s).
HRMS(ESI):m/z[M+H]
+calcd?for?C
36H
60NO
3:554.4573；found:554.4579。
O-(morpholinyl) the ethyl derivative antibacterial activity of embodiment 4 Cleistanone
The anti-human body fungi activity of O-(morpholinyl) ethyl derivative (III) of experimental example 1 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Anti-human body fungi activity experiment is the method that adopts concentration dilution, each mensuration in triplicate, and the pathogen of test has trichophyton, microsporum canis and trichophyton tonsurans, and bacterial concentration is 10
5individual/mL.O-(dimethylamino) ethyl derivative (III) initial concentration of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is 50.00 μ g/mL (5% dimethyl sulfoxide DMSO), gradient dilution to 0.10 μ g/mL, (ultimate density of formation has: 25.00 in 96 orifice plates for the bacterium liquid 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 ℃, after incubation time 24h, observe, if find, there is no that minimum concentration in processing hole that bacterium colony forms is sample lowest concentration of antimicrobial, it is MIC value.This experiment positive control is ketoconazole, and the anti-Human Fungi of chemicals III the results are shown in Table 1.
The anti-Human Fungi MIC of O-(morpholinyl) ethyl derivative (III) value (μ g/mL) of table 1 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Conclusion: O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has very strong anti-human body fungi activity, O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone therefore of the present invention is expected to be used to prepare novel anti-human body fungi-medicine.
O-(morpholinyl) ethyl derivative (III) anti-helicobactor pylori activity of experimental example 2 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
1) strains tested
Helicobacter pylori (Helicobacter pylori, Hp) reference culture ATCC43504 is purchased from U.S.'s strain and preserves center (American Type Culture Collection, ATCC).13 strain Hp clinical strains are picked up from Nanjing drum tower hospital Dndoscope Laboratory and are accepted gastroscopic patient; Patient to peptic ulcer, duodenal bulbar inflammation or gastritis verrucosa in continuous gastroscopy, first through RUT experiment, be defined as Hp positive, get again antral gastric mucosa 1-2 piece, after chopping, be inoculated in containing 8% horse serum, trimethoprim (trimethropin, TMP) in the Columbia selectivity agar culture medium of 1.25g/L, polymyxin (polymyxin) B2500U/L, vancomycin (vancomycin) 10mg/L, in 37 ℃ of (5%O under micro-oxygen environment
2, 10%CO
2and 85%N
2) cultivate 72 hours.Collect antibacterial, through smear Gram’s staining, oxidase, catalase and urease are accredited as after the positive, the pure culture of going down to posterity, and obtained strains is as experimental strain.
2) strain culturing
We adopt micro-aerobic bag (purchased from Shanghai Medical Univ) to carry out the strain culturing of HP, and it can produce the needed micro-aerobic environment of Hp by chemical reaction.
3) biological activity determination
Adopt paper disk method (Microbiological paper method) to measure the inhibitory action of compound to helicobacter pylori, with agar dilution, measure the minimum inhibitory concentration (minimal inhibitory concentration, MIC) of test sample.
I. paper disk method experiment
(A) prepare culture medium by the Columbia culture medium preparing after high pressure steam sterilization, be cooled to 50-60 ℃, add 8% horse serum or Sheep Blood, mix in the culture dish that is poured into sterilizing, every ware 7-10ml, culture medium thickness is 1.5mm (sterile working).
(B) switching experimental bacteria (be coated with bacterium) with microscale sampler get diluted 10
8cFU/ml (1OD
660=10
8cFU/ml) the bacteria suspension 0.1ml of Hp spreads upon suitable culture dish surface equably.Be inverted in 37 ℃ of drying bakers and take out after 15min, object makes agar surface dry, standby.
(C) pasting the sample scraps of paper gets 6 μ l testing samples (mass concentration 2mg/ml) with microscale sampler and injects on the round filter paper of sterilizing.With aseptic nipper tweezer containing the scraps of paper of sample and the blank scraps of paper of contrast, by sterile working respectively the scraps of paper be close to containing bacterio-agar surface, paste at a certain distance a piece of paper sheet.Every kind of bacterium is cooked 3 wares, and acquired results is asked its meansigma methods.
(D) cultivate each plate is placed in to micro-aerobic bag, sealing, opens gas generator, then is placed in 37 ℃ of incubators and cultivates 72h.
(E) survey antibacterial circle diameter and take out after flat board, measure respectively each scraps of paper size of antibacterial circle diameter around.With reference to the result of matched group, can draw the result of testing sample sensitive experiment.In triplicate.
II. agar dilution is measured MIC
(A) first the preparation of medicine flat board becomes dimethyl sulfoxide for compound (DMSO) solution preparation of test the mother solution of 0.5mg/ml, then with sterilized water dilution, is finally made into 100.0,80.0,60.0,45.0,40.0,35.0,30.0,25.0,20.0,15.0,10.0, the concentration series of 5.0 and 2.5 μ g/ml, the concentration of DMSO in medium is less than 1%.The test compounds solution that 1ml is prepared be incubated the 8ml Colombia culture medium in 50 ℃, separately add 1ml and be incubated in the horse serum of 50 ℃ and fully mix, cast that cooling in culture dish (in culture dish, the final concentration of chemicals is 10.0,8.0,6.0,4.5,4.0,3.5,3.0,2.5,2.0,1.5,1.0,0.5 and 0.25 μ g/ml, if there is bacteriostasis, MIC value is these in worthwhile).
(B) switching experimental bacteria (be coated with bacterium) with microscale sampler draw diluted 1 * 10
8the bacteria suspension 0.1ml of CFU/ml Hp spreads upon culture dish surface equably, is inverted in 37 ℃ of drying bakers and takes out after 15min, and object makes agar surface dry, standby.
(C) determine that MIC (contains culture dish to be measured: 85%N at micro-aerobic bag
2, 10%CO
2and 5%O
2) in, be incubated 37 ℃ and cultivate 72 hours, observe Hp growing state, with blank group contrast, take and there is no the sample of bacteria growing least concentration completely as minimum inhibitory concentration (minimal inhibitory concentration, MIC) value.Positive control is ampicillin (Ampicillin).
Experimental result
Experiment in vitro shows, O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has very strong anti Helicobacter pylori activity, and paper disk method shows that its antibacterial circle diameter is 23mm (ATCC43504).With agar dilution, show that it can suppress the growth of 5 random clinical strains and 1 reference culture (ATCC43504) completely, minimal inhibitory concentration (MIC) is 2.0 μ g/ml.With ampicillin, make positive control, its Cmin (MIC) that 6 strain test bacterium are suppressed is completely 2.5 μ g/ml.
Conclusion: it is active that O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has stronger inhibition helicobacter pylori, explanation for the disease such as the closely-related acute and chronic gastritis of helicobacter pylori, duodenal ulcer, O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is the compound of a great exploitation potential for its.It can be directly used in the treatment of corresponding disease and the preparation of related drugs.
O-(morpholinyl) ethyl derivative (III) antibacterial activity of experimental example 3 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Antibacterial activity test is the method that adopts concentration dilution, each mensuration in triplicate, and test pathogen has escherichia coli, fluorescent pseudomonas, Staphylococcus aureus, Bacillus proteus, neogenesis cryptococcus, and bacterial concentration is 10
5individual/mL.O-(morpholinyl) ethyl derivative (III) initial concentration of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is 50.00 μ g/mL (5% dimethyl sulfoxide DMSO), gradient dilution to 0.10 μ g/mL, (ultimate density of formation has: 25.00 in 96 orifice plates for the bacterium liquid 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), antibacterial culturing temperature is respectively 37 ℃, after incubation time 24h, observe, if find, there is no that minimum concentration in processing hole that bacterium colony forms is sample lowest concentration of antimicrobial, it is MIC value.O-(morpholinyl) ethyl derivative (III) the anti-bacterial result of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is in Table 2.
O-(morpholinyl) ethyl derivative (III) the antibacterium MIC value (μ g/mL) of table 2 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Conclusion: O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has very strong antibacterial activity, O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone therefore of the present invention is expected to be used to prepare novel anti-bacterial drug.
O-(morpholinyl) ethyl derivative (III) anti-tubercle bacillus of experimental example 4 Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is active
(1) the anti-bacillus calmette-guerin vaccine of O-(morpholinyl) ethyl derivative (III) (BCG) absolute concentration of solid medium By Dilution Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Scraping bacillus calmette-guerin vaccine culture from inclined-plane, join in 3ml Middlebrook7H9 broth bouillon, add a small amount of bead, screw test tube cap, on vortex oscillation device, high vibration grinds, than turbid, be made into bacillus calmette-guerin vaccine (BCG) bacteria suspension of 1mg/ml with standard Maxwell opacity tube (MacFarland No.1).
O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is made into the stock solution of high concentration with DMSO, with the aseptic ultra-pure water of the tween 80 containing 5%, dilute stock solution to desired concn, O-(morpholinyl) ethyl derivative (III) of the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone having diluted is joined to 4mlMiddlebrook7H11 agar culture medium by required dosage, and (this culture medium is 121 ℃ of high pressure steam sterilizations 15 minutes, be cooled to 50～55 ℃), mix, make O-(morpholinyl) ethyl derivative (III) containing Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, 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 bacillus calmette-guerin vaccine that is 1mg/ml by concentration (BCG) bacteria suspension dips ring of numbers with inoculating loop, be inoculated in respectively in the culture medium and blank medium slant containing O-(morpholinyl) ethyl derivative (III) series concentration of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, being placed in 37 ℃ cultivates 4～8 weeks, observation experiment result, result is as shown in table 3.
Conventional culture medium when Middlebrook7H9 broth bouillon used and Middlebrook7H11 agar culture medium carry out tulase cultivation for those skilled in the art in the present embodiment, its formula adopts conventional formulation.
(2) O-(morpholinyl) ethyl derivative (III) the Killing Mycobacterium Tuberculosis type strain H37Rv strain absolute concentration of solid medium By Dilution Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Scraping mycobacterium tuberculosis type strain H37Rv strain culture from inclined-plane, join in 3mlMiddlebrook7H9 broth bouillon, add a small amount of bead, screw test tube cap, on vortex oscillation device, high vibration grinds, than turbid, be made into the H37Rv strain bacteria suspension of 1mg/ml with standard Maxwell opacity tube (MacFarland No.1).
O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is made into respectively to the stock solution of high concentration with DMSO, with the aseptic ultra-pure water of the tween 80 containing 5%, dilute stock solution to desired concn, O-(morpholinyl) ethyl derivative (III) of the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone having diluted is joined to 4mlMiddlebrook7H11 agar culture medium by required dosage, and (this culture medium is 121 ℃ of high pressure steam sterilizations 15 minutes, be cooled to 50～55 ℃), mix, make O-(morpholinyl) ethyl derivative (III) containing Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, 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 that is 1mg/ml by concentration dips ring of numbers with inoculating loop, be inoculated in respectively in the culture medium and blank medium slant containing O-(morpholinyl) ethyl derivative (III) series concentration of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, being placed in 37 ℃ cultivates 4～8 weeks, observation experiment result, result is as shown in table 3.
(3) the clinical separation of O-(morpholinyl) ethyl derivative (III) the tuberculosis mycobacterium MTB of the resistance to ISRE strain absolute concentration of solid medium By Dilution Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
The clinical separation of the scraping mycobacterium tuberculosis MTB of resistance to ISRE strain (resistance to isoniazid, streptomycin, rifampicin, the clinical detached dowel of ethambutol mycobacterium tuberculosis) culture from inclined-plane, join in 3ml Middlebrook7H9 broth bouillon, add a small amount of bead, screw test tube cap, on vortex oscillation device, high vibration grinds, than turbid, be made into the bacteria suspension of 1mg/ml with standard Maxwell opacity tube (MacFarland No.1).
O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone is made into respectively to the stock solution of high concentration with DMSO, with the aseptic ultra-pure water of the tween 80 containing 5%, dilute stock solution to desired concn, O-(morpholinyl) ethyl derivative (III) of the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone having diluted is joined to 4mlMiddlebrook7H11 agar culture medium by required dosage, and (this culture medium is 121 ℃ of high pressure steam sterilizations 15 minutes, be cooled to 50～55 ℃), mix, make O-(morpholinyl) ethyl derivative (III) containing Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, 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 clinical separation of the mycobacterium tuberculosis MTB of the resistance to ISRE strain bacteria suspension that is 1mg/ml by concentration dips ring of numbers with inoculating loop, be inoculated in respectively in the culture medium and blank medium slant containing O-(morpholinyl) ethyl derivative (III) series concentration of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone, being placed in 37 ℃ cultivates 4～8 weeks, observation experiment result, result is as shown in table 3.
O-(morpholinyl) ethyl derivative (III) the anti-tubercle bacillus absolute concentration result of table 3 solid medium By Dilution Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone
Conclusion: O-(morpholinyl) ethyl derivative (III) of Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone has very strong anti-tubercle bacillus and anti-drug resistance tulase is active, can be used as the lead compound for the treatment of tubercle bacillus affection disease, also can be used for preparation treatment tuberculosis medicine.
The preparation of O-(morpholinyl) the ethyl derivative tablet of embodiment 5 Cleistanone involved in the present invention
Get a kind of in the middle of O-(morpholinyl) ethyl derivative of 20 grams of Cleistanone or its pharmaceutically acceptable salt, add 180 grams of conventional adjuvants preparing tablet, mix, conventional tablet machine is made 1000.
The preparation of O-(morpholinyl) the derivatized composite capsule of embodiment 6 Cleistanone involved in the present invention
Get a kind of in the middle of O-(morpholinyl) ethyl derivative of 20 grams of Cleistanone or its pharmaceutically acceptable salt, add the conventional adjuvant of preparing capsule as 180 grams of starch, mix, encapsulatedly make 1000.
Claims (8)
1. O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III:
2. a kind of O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in preparation antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III as claimed in claim 1, is characterized by: described bacterium is antibacterial.
3. a kind of O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III as claimed in claim 1, is characterized by: described bacterium is fungus.
4. a kind of O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III as claimed in claim 1, is characterized by: described bacterium is helicobacter pylori.
5. a kind of O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III as claimed in claim 1, is characterized by: described bacterium is tulase.
6. a kind of O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III as claimed in claim 3, is characterized by: described fungus is trichophyton, microsporum canis or trichophyton tonsurans.
7. a kind of O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III as claimed in claim 2, is characterized by: described antibacterial is escherichia coli, fluorescent pseudomonas, Staphylococcus aureus, Bacillus proteus or neogenesis cryptococcus.
8. a kind of O-(morpholinyl) ethyl derivative and the application of pharmaceutically acceptable salt in antibacterials thereof with the Cleistanthus sumafranus (Miq) Muell-Arg-C. Saichikii Merr wood ketone Cleistanone of structure shown in formula III as claimed in claim 5, is characterized by: described tulase is bacillus calmette-guerin vaccine, the H37Rv strain of mycobacterium tuberculosis type strain and substance of medicines-resistant branched tubercle bacillus.
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| CN104758296A (en) * | 2015-03-10 | 2015-07-08 | 江苏卓见医疗用品有限公司 | Chitosan composite material containing novel active molecule, preparation method and application thereof |
| CN104840470A (en) * | 2015-04-15 | 2015-08-19 | 南京广康协生物医药技术有限公司 | Application of cleistanone O-(1H-tetrazole)ethyl derivative in preparation of antibacterial drugs |
| CN104873509A (en) * | 2015-05-27 | 2015-09-02 | 南京大学 | Application of O-(diethylamino) ethyl derivative of Daphmalenine A in preparation of antibacterial drug |
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Cited By (3)
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