CN109232684A - 17-AAG glucoside and preparation method thereof and application in preparation of anti-tumor drugs - Google Patents
17-AAG glucoside and preparation method thereof and application in preparation of anti-tumor drugs Download PDFInfo
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Abstract
The present invention is using 17-AAG as substrate, method is glycosylated by external enzyme process and prepares novel Corylifol A glycosylated derivative 17-AAG glucoside using UDP-glucose as donor using glycosyl transferase (YjiC), shown in its structural formula such as formula (I)And study its physicochemical property, it was demonstrated that the water solubility of 17-AAG glucoside is water-soluble 10.5 times of substrate 17-AAG, and has preferable stability to pH；And 17-AAG glucoside inhibits the active IC of Hsp90ATPase50Value is 131.8 μm of ol/L；All there is very strong Inhibit proliferaton effect to human hepatocarcinoma BEL-7402, human hepatoma HepG2 cell, human breast carcinoma MDA-MB-231 cell, human colon carcinoma SW480 cell in vitro, and also turn out that 17-AAG glucoside also has the activity for significantly inhibiting tumour growth in vivo through nude mice in vivo studies, inhibiting rate is 51.8% when being administered 21 days, and toxicity is low, with it is current clinic used in antineoplastic drug compared with, with good clinical antineoplastic medicine application prospect.
Description
Technical field
The invention belongs to pharmaceutical technology field, it is specifically related to a kind of 17-AAG glucoside and preparation method thereof and is making
Application in standby anti-tumor drug.
Background technique
In recent years, Several Kinds of Malignancy disease incidence is worldwide in that rising is taken advantage of a favourable situation, seriously endanger human health and
Life, and chemotherapy is one of the important means for the treatment of malignant tumour.Therefore, safe and efficient antitumor innovation is provided to clinic
Drug is the important mission of current medical personal.
Heat shock protein 90 (Hsp90) adjusts the conformational maturation and stabilization of numerous signal proteins as molecular chaperone protein
Property, cell growth, differentiation, apoptosis, canceration and tumour in terms of play an important role, it has also become antineoplastic
One of the important target spot of object research and development.Currently, more than 200 kinds albumen are confirmed as the client protein of Hsp90, including cross-film
Tyrosine kinase receptor (Her-2), vascular endothelial growth factor receptor, Akt, Raf-1, jump signal albumen (v-Src, p53),
HIF-1 α, MMPs etc. play an important role in tumour generation, the signal path shifted.
Geldanamycin (geldanamycin) is first Hsp90 inhibitor being found, and is initially from water suction strepto-
Isolated natural products, belongs to benzoquinones ansha antibiosis in bacterium (Streptomyces hygroscopicus) fermentation liquid
Element.Geldanamycin has very strong anti-tumor activity as antitumor drug candidate, but due to its hepatotoxicity wind agitation, poorly water-soluble etc.
Disadvantage limits application clinically.In recent years, many scholars be improve geldanamycin water solubility, reduce its toxicity and
Enhance its bioavilability and compare extensive structure of modification, and achieves certain achievement.Such as, 17-AAG
The anti-tumor activity of (Tanespimycin, the analog of geldanamycin) is higher than geldanamycin, and hepatotoxicity wind agitation is also significantly
It reduces, has come into the III clinical trial phase stage at present.But 17-AAG still there are certain hepatotoxicity wind agitation, it is water-soluble compared with
The deficiencies of difference, limited bioavailability.
Small molecule compound it is glycosylation modified, can not only increase compound structure diversity, and functionally this
A little glycan molecules have also assisted in the identification of aim cell, improve compound and obtain water-soluble, have influenced bioactivity.Such as 7 β-
Xylosyl-10-deacetyl taxol (CAS:90332-63-1) and its derivative are shown while retaining anti-tumor activity
It lands and improves its specific selectivity water-soluble and to tumour cell.Currently used glycosylation approach has chemical synthesis
Method, biosynthesis pathway engineering technology, conversion technology and external enzyme process glycosylation (in vitro enzymatic
Glycosylation) etc..Although external enzyme process glycosylation has satisfactory yield and selectivity, one kind is found
Glycosyl transferase with substrate specificity selectivity is key, and glycosyl transferase is in glycosylated natural products GCMS computer approach
A kind of important enzyme, major function are that the sugar being catalyzed on donor is transferred to non-saccharide body (aglycon).
Summary of the invention
The purpose of the present invention is to provide a kind of 17-AAG glucoside and preparation method thereof and preparing anti-tumor drug
In application using UDP-glucose as saccharide donor, structure is carried out to substrate 17-AAG and is repaired using glycosyl transferase (YjiC)
Decorations, prepare 17-AAG glucoside by external enzyme process glycosylation, obtain satisfactory yield and selectivity, and
Study its physicochemical property, it was demonstrated that 17-AAG glucoside of the invention not only has preferable water-soluble and stability, also has
Significant antitumor action, 17-AAG glucoside and its salt have broad application prospects in the preparation of antitumor drugs.
To achieve the above object, the present invention provides a kind of 17-AAG glucosides, have the structure as shown in formula (I)
Formula:
The preparation method of 17-AAG glucoside of the invention, specifically:
Tris-HCl, 1mmol/L MgCl for being 6.0~9.6 by pH value containing 25mmol/L2, 25mg YjiC, 3~
The reaction solution 50mL of 6mmol/L UDP-glucose and 3mmol/L 17-AAG react 6~9 hours at 30 DEG C；After reaction,
100 DEG C heating water bath 10 minutes, eliminate the activity of glycosyl transferase, then into reaction solution be added 50mL ethyl acetate to reaction solution
It is extracted, obtains ethyl acetate extract, through half preparation efficient liquid phase purification, obtain 17-AAG glucoside.
The present invention to the specific selectivity of 17-AAG, is supplied using glycosyl transferase (YjiC) using UDP-glucose as sugar
Body prepares 17-AAG glucoside by external enzyme reaction, and the 17-AAG glucoside prepared not only has good water
Dissolubility and stability also have significant antitumor action.
The present invention also provides 17-AAG glucoside application in preparation of anti-tumor drugs, the tumour cell is
The tumour cell of digestive system, gastrointestinal system and reproductive system position, especially liver cancer cells, colon cancer cell and breast cancer are thin
Born of the same parents.
The water-soluble of 17-AAG glucoside of the invention increases by 10.5 times than the water-soluble of substrate 17-AAG, and to pH
With better stability；While 17-AAG glucoside inhibits Hsp90 ATPase activity in vitro, suppression also can be effectively suppressed
Human liver cancer SMMC7721 cell processed, human hepatoma HepG2 cell, human breast carcinoma MDA-MB-231 cell and human colon carcinoma SW480 are thin
The proliferation of born of the same parents, and concentration dependent is presented, IC50Value is respectively 5.26,6.28,28.52,6.13 μm of ol/L.It is moved in nude mice
It plants in tumor model, shows preferable anti-tumor activity, there is good potential applicability in clinical practice.
The 17-AAG glucoside or its salt and drug excipient or vehicle group are at pharmaceutical composition, pharmaceutical composition
Dosage form include liquid preparation, tablet, granule, electuary, capsule and pill, capsule, sustained release agent, pill or oral cavity disintegration preparation.
Detailed description of the invention
Fig. 1 is the pH stability of 17-AAG glucoside.
Fig. 2 is influence of the 17-AAG glucoside to customer's Hsp90 protein expression.
Fig. 3 is the internal anti-tumor activity of 17-AAG glucoside.
Specific embodiment
Invention is further illustrated with reference to embodiments.
17-AAG glucoside, structural formula are as follows:
The preparation method of embodiment 1,17-AAG glucoside, specifically:
Experimental material:
Instrument: half preparative high-performance liquid chromatographic instrument (Waters, US), Bruker ARX Nuclear Magnetic Resonance (Germany
Bruker company), JMS-HX/HX110A type mass spectrograph (Japanese JEOL company).
Reagent: acetonitrile, methanol (Fisher company of the U.S.).
External enzyme process glycosylase reaction:
By the MgCl of Tris-HCl, the 1mmol/L for being 8.8 of pH containing 25mmol/L2, 25mg YjiC, 6mmol/L UDP-
The reaction solution 50mL of glucose and 3mmol/L 17-AAG reacts 6 hours at 30 DEG C, after reaction, in 100 DEG C of heating water baths
10 minutes, the activity of glycosyl transferase is eliminated, then 50mL ethyl acetate is added to reaction solution and extracts to reaction solution, obtain second
Acetoacetic ester extract 80mg, through half preparation efficient liquid phase (Waters 2535Q system；Chromatographic column is SunfireTM prep
C18,10x250mm；Mobile phase is 40% acetonitrile；Flow velocity is 4mL/min) purification, obtain 17-AAG glucoside 20.6mg.
Structural Identification:
17-AAG glucoside is brown solid；HR-ESI-MS m/z[M-H]-746.3506 [M+Na]+770.3487
Molecular formula is C37H53N3O13.In addition, 17-AAG glucoside is dissolved in DMSO-d6, carried out magnetic resonance detection, the Portugal 17-AAG
The NMR data of polyglycoside are as follows: δH 10.18(17-NH,s),9.39(1-NH,s),7.06(1H,s,H-3),6.82(1H,s,H-
19), 6.54 (1H, t, J=10.2Hz, H-4), 5.90 (1H, m, H-27), 5.74 (1H, dd, J=10.2,5.7Hz, H-5),
5.45(1H,m,H-9),5.23(2H,m,H-7,H-28),5.11(1H,m,H-28),5.23,m4.54(1H,m,H-6),4.23
(1H, d, J=7.5Hz, H-1 '), 4.09 (2H, m, H-26), 3.83 (m, H-6 '), 3.60 (1H, m, H-12), 3.39 (m, H-
6′),3.20(3H,s,12-OCH3),3.16(3H,s,6-OCH3),3.11(m,H-3′),3.02(1H,m,H-11),2.95(m,
H-2′),2.79(m,H-4′,H-5′),2.65(1H,m,H-10),1.98(1H,m,H-14),1.91(3H,s,H-22),1.60
(3H, s, H-24), 1.58 (2H, m, H-15), 1.40 (2H, m, H-13), 1.03 (3H, d, J=6.8Hz, H-23), 0.84 (3H,
D, J=6.8Hz, H-25)；δC184.59(C-18),178.9(C-21),169.43(C-1),156.72(7-OCONH2),
146.11(C-17),141.34(C-20),138.34(C-5),135.74(C-27),133.86(C-2,C-8),130.36(C-
3),129.16(C-9),125.92(C-4),116.18(C-28),109.06(C-16),107.56(C-19),103.46(C-
1′),81.58(C-12),79.16(C-7),79.12(C-6),78.94(C-11),77.42(C-3′),76.91(C-5′),
74.42(C-2′),70.98(C-4′),61.93(C-6′),56.71(12-OCH3),56.41(6-OCH3),46.7(C-26),
37.8(C-13),33.06(C-10),33.3(C-15),30.03(C-14),21.22(C-25),16.8(C-24),14.4(C-
22),12.67(C-23)。
The preparation method of embodiment 2,17-AAG glucoside, specifically:
By the MgCl of Tris-HCl, the 1mmol/L for being 6.0 of pH containing 25mmol/L2, 25mg YjiC, 6mmol/L UDP-
The reaction solution 50mL of glucose and 3mmol/L 17-AAG reacts 9 hours at 30 DEG C, after reaction, in 100 DEG C of heating water baths
10 minutes, the activity of glycosyl transferase is eliminated, then 50mL ethyl acetate is added to reaction solution and extracts to reaction solution, obtain second
Acetoacetic ester extract 80mg, through half preparation efficient liquid phase (Waters 2535Q system；Chromatographic column is SunfireTM prep
C18,10x250mm；Mobile phase is 40% acetonitrile；Flow velocity is 4mL/min) purification, obtain 17-AAG glucoside 17.8mg.
The preparation method of embodiment 3,17-AAG glucoside, specifically:
By the MgCl of Tris-HCl, the 1mmol/L for being 9.6 of pH containing 25mmol/L2, 25mg YjiC, 3mmol/L UDP-
The reaction solution 50mL of glucose and 3mmol/L 17-AAG reacts 9 hours at 30 DEG C, after reaction, in 100 DEG C of heating water baths
10 minutes, the activity of glycosyl transferase is eliminated, then 50mL ethyl acetate is added to reaction solution and extracts to reaction solution, obtain second
Acetoacetic ester extract 80mg, through half preparation efficient liquid phase (Waters 2535Q system；Chromatographic column is SunfireTM prep
C18,10x250mm；Mobile phase is 40% acetonitrile；Flow velocity is 4mL/min) purification, obtain 17-AAG glucoside 17.6mg.
The preparation method of embodiment 4,17-AAG glucoside, specifically:
By the MgCl of Tris-HCl, the 1mmol/L for being 6.0 of pH containing 25mmol/L2, 25mg YjiC, 3mmol/L UDP-
The reaction solution 50mL of glucose and 3mmol/L 17-AAG reacts 6 hours at 30 DEG C, after reaction, in 100 DEG C of heating water baths
10 minutes, the activity of glycosyl transferase is eliminated, then 50mL ethyl acetate is added to reaction solution and extracts to reaction solution, obtain second
Acetoacetic ester extract 80mg, through half preparation efficient liquid phase (Waters 2535Q system；Chromatographic column is SunfireTM prep
C18,10x250mm；Mobile phase is 40% acetonitrile；Flow velocity is 4mL/min) purification, obtain 17-AAG glucoside 11.9mg.
Embodiment 5, water-soluble detection
Instrument: KQ 5200B Ultrasound Instrument (Chinese Kunshan ultrasonic instrument Co., Ltd)；Centrifuge (Italian ALC company)；
HPLC (Dionex company of the U.S.).
Reagent: acetonitrile, methanol (Fisher company of the U.S.).
Method: taking the 17-AAG glucoside of sufficient amount 17-AAG and preparation, is dissolved in respectively equipped with 600 μ L distilled water
In 1.5mL centrifuge tube, and make it in hypersaturated state using ultrasonic wave auxiliary dissolution at room temperature, ultrasonic wave dissolves 40 minutes
Afterwards, then 10000r/min is centrifuged 20 minutes, takes supernatant to be diluted to debita spissitudo, injection HPLC is analyzed, and is asked water-soluble
The concentration of compound in liquid.
It is 493.58 μ g/mL that experimental result, which is shown in Table the solubility in the water of 1:17-AAG glucoside, is substrate 17-AAG
10.5 times.
The water-soluble testing result of table 1.
Embodiment 6, the Detection of Stability to pH
Instrument: Mini Dry Bath (Hangzhou meter Ou Instrument Ltd.).
Reagent: acetonitrile, methanol (Fisher company of the U.S.).
Method: by the 17-AAG glucoside of preparation and substrate 17-AAG be dissolved in respectively 200 μ L difference pH (6.0,7.0,
8.0, it 8.8,9.6) in Tris-HCl buffer, reacts 30 minutes at room temperature, and inject HPLC and analyzed.
Experimental result indicates its stability with the ratio of former compound as shown in Figure 1:, under the conditions of certain pH, 17-
The product 17-AAG glucoside that AAG is modified through glycosylation structure is more stable than substrate 17-AAG.
Embodiment 7, the ATPase Activity determination to Hsp90
Instrument: multi-function microplate reader (U.S. BioTek).
Method: malachite green-molybdat chromogenic reaction is used.By the Hsp90-his fusion protein of high-purity
2 μm of ol/L are placed in 96 orifice plates, and compound (17-AAG glucoside, 17-AAG and the Ge Erde of various concentration is added in each hole
Mycin) and reaction buffer (100mmol/L Tris-HCl, 20mmol/L KCl, 6mmol/L MgCl2, pH=7.4) and it mixes
It is placed on 37 DEG C to react 2.5 hours, each compound concentration sets three multiple holes.80 μ L malachite are added after reaction
Green-molybdate solution (CAS:68083-41-0) colour developing, and absorbance (A) value in each hole is surveyed in wavelength 620nm, and
Compound is calculated to the ATPase inhibitory activity of Hsp90, the above experiment is repeated 3 times.
Experimental result is shown in Table 2:17-AAG glucoside and inhibits the active IC of Hsp90ATPase50Value is 131.8 μm of ol/L,
And the IC of positive control drug geldanamycin50Value is 3.2 μm of ol/L.
2 17-AAG glucoside of table inhibits Hsp90 ATPase Activity determination result
| Hsp90 inhibitory activity (IC50, μm ol/L) | |
| 17-AAG glucoside | 131.8±10.5 |
| 17-AAG | 31.9±2.9 |
| Geldanamycin | 3.2±0.2 |
The influence of embodiment 8, mtt assay detection 17-AAG glucoside to four kinds of tumor cell proliferations
The experimental section by human liver cancer SMMC7721 cell, human hepatoma HepG2 cell, MCF-7 Human Breast Cancer Cells,
The lethal effect of human colon carcinoma SW480 cell carries out effect assessment.
(1) experimental material:
Cell strain: human liver cancer SMMC7721 cell, human hepatoma HepG2 cell, MCF-7 Human Breast Cancer Cells, human colon carcinoma
SW480 cell origin is in the U.S. American Type Culture Collection (ATCC) company.
Instrument: carbon dioxide incubator (SHELL LAB S. A. of the U.S.), multi-function microplate reader (U.S. BioTek) are inverted
Microscope (Japanese OLYMPUS company).
Reagent: thiazolyl blue (MTT) is purchased from Sigma Co., USA；Culture solution, dimethyl sulfoxide (DMSO), 0.25% pancreas egg
White enzyme, penicillin and streptomysin are purchased from Hyclone；96 well culture plates are purchased from Corning company；Fetal calf serum is purchased from Hangzhou China
Chinese holly biotech company.
(2) method:
Cell culture: four kinds of tumour cells are inoculated in DMEM or RPMI1640 culture solution respectively (containing 10% inactivation tire
Cow's serum, 100IU/l penicillin, 100 μ g/mL streptomysins), set 5%CO2, 37 DEG C, cultivate and pass under saturated humidity environment.
Mtt assay: the cell of logarithmic growth phase is made single cell suspension with 0.25% trypsin digestion, is inoculated in 96
In orifice plate, 5000 cells are contained in every 100 μ L culture solution of hole, in 5%CO2It is cultivated in 37 DEG C of incubators of saturated humidity.Culture
After 14 hours, compound (while setting positive controls, 20 μm of ol/L of geldanamycin) is handled by various concentration, each processing 3
A multiple holes, the MTT solution that every hole is added that 10 μ L concentration are 5g/L after continuing culture 72 hours continue to be incubated for 4 hours, discard culture
150 μ L of DMSO is added in liquid, every hole, and 37 DEG C are incubated for 30 minutes, and micro oscillator, which vibrates 10 minutes, dissolves crystal sufficiently, use
Microplate reader detects absorbance (A) value in every hole under 570nm wavelength, calculates cell survival rate (cell survival rate/%=experimental group
A570nm/ control group A 570nm × 100%), dose-effect curve is drawn, the above experiment is repeated 3 times.
(3) experimental result is shown in Table shown in 3, and 17-AAG glucoside is in vitro to human hepatocarcinoma BEL-7402, human liver cancer
HepG2 cell, human breast carcinoma MDA-MB-231 cell, human colon carcinoma SW480 cell have very strong Inhibit proliferaton effect, IC50
Value is respectively 5.26,6.28,28.52,6.13 μm of ol/L.
Influence of the 3 17-AAG glucoside of table to four kinds of tumor cell proliferations
Embodiment 9,17-AAG glucoside are to Akt, HIF-1 α in human hepatocarcinoma BEL-7402 and Hsp90 albumen table
The influence reached
(1) experimental material:
Cell strain: human hepatocarcinoma BEL-7402 derives from U.S. American Type Culture Collection
(ATCC) company.
Instrument: carbon dioxide incubator (SHELL LAB S. A. of the U.S.), inverted microscope (Japanese OLYMPUS company) coagulate
Glue imaging system (BIO-RAD company of the U.S.)；Electrophoresis apparatus (BIO-RAD company of the U.S.)；Inverted fluorescence microscope (Japan
OLYMPUS company)；Ultracentrifuge (U.S. Beckman).
Reagent: DMEM culture solution, DMSO, 0.25% trypsase, penicillin and streptomysin are purchased from Hyclone；6 orifice plates purchase
From Corning company；Fetal calf serum is purchased from Hangzhou China Chinese holly biotech company；Rabbit-anti people Akt antibody (U.S. Abcam
Company), rabbit-anti people HIF-1 Alpha antibodies (Abcam company of the U.S.), Goat anti-Human Hsp90 Alpha antibodies (Stressgen company of the U.S.),
β-actin (U.S. Santa Cruz)；Secondary antibody (Chinese Hefei BioSharp company)；ECL reaction solution (U.S. Santa Cruz).
(2) method:
It is 2X10 that human hepatocarcinoma BEL-7402, which is pressed concentration,5/ mL is inoculated in 6 orifice plates, and the hole 2mL/ is pasted completely to cell
Wall post-processes compound, continues culture 24 hours.Culture cell is collected, after being cleaned with ice PBS, filter paper is added carefully after blotting liquid
Cellular lysate liquid, 200 holes μ L/ are set 30 minutes on ice, under cell scraper, are transferred in EP pipe and are frozen for 80 DEG C, after 3 multigelations
BCA standard measure.40 μ g of protein extract is taken, 5XSDS sample-loading buffer is added, 100 DEG C are boiled 10 minutes, and SDS-PAGE electricity is done
Swimming, spacer gel constant pressure 50V, separation gel constant pressure 100V, electrophoresis to bromophenol blue fuel reach gel forefront, stop electrophoresis.
Transferring film: glue is taken off after electrophoresis, and gel is dipped in suitable Transfer buffer.It takes simultaneously appropriate big
PVDF is first soaked in anhydrous methanol, is then dipped in Transfer together with filter paper by small pvdf membrane and 4 3M filter paper
In buffer, 2 3M filter paper are respectively padded on the two sides of film, constant pressure 60V, transferring film 2 hours in 4 DEG C of chromatography cabinets.
Sealer: the film for turning to have albumen is dipped in the TBST containing 10% skimmed milk power and is closed 1 hour.Hybridization: taking-up has been sealed
Then the film closed is dipped in the diluted rabbit-anti people Akt antibody of 1:500~1:1000, rabbit-anti people HIF-1 Alpha antibodies, Goat anti-Human
In Hsp90 Alpha antibodies (preparation of the TBST containing 5% skimmed milk power, pH 7.4), overnight at 4 DEG C.TBST is rinsed 5 minutes X 5 times, then
It is dipped in the diluted secondary antibody of 1:2000 (TBST containing 5% skimmed milk power, pH7.4 are prepared), at room temperature 1 hour, TBST rinsed 5 points
Clock X 4 times.It prepares developing solution (ECL A 0.5mL, ECL B 0.5mL), places chromogenic assay in gel imaging system.
(3) experimental result as shown in Fig. 2, with 17-AAG glucoside concentration increase, Akt and HIF-1 alpha protein contains
Amount substantially reduces.Swimming lane 1: blank control group；The 17-AAG glucoside of 2:5 μm of ol/L of swimming lane；The 17- of 3:10 μm of ol/L of swimming lane
AAG glucoside；The 17-AAG glucoside of 4:20 μm of ol/L of swimming lane.17-AAG glucoside is in 5~20 μm of ol/L concentration
Dose-dependently inhibit the expression of Akt albumen in SMMC-7721 cell, and significantly inhibits HIF- when 20 μm of ol/L administration concentrations
The expression of 1 α albumen.At the same time, 17-AAG glucoside is in 5~20 μm of ol/L concentration, Hsp90 α in SMMC-7721 cell
Expressing quantity does not change.
The internal anti-tumor activity detection of embodiment 10,17-AAG glucoside
(1) method: purchase BALB/c strain nude mice, 4~5 week old, 18~20g of weight, in Bengbu Medical College experimental animal
It is tested at center (SPF grades).The good SMMC-7721 cell of growth conditions is taken to be digested, be centrifuged, counted, with sterile PBS
Buffer is deployed into 6X107/ mL suspension injects 100 μ L cell suspensions in the back channel injected s.c. of every mouse.
After tumour cell is successfully inoculated with, gross tumor volume reaches 100mm3When left and right, nude mice is grouped into blank control group, administration group
(17-AAG glucoside, 10mg/kg), every group of 3 nude mices, administration mode are intraperitoneal injection, and the period is 3 days primary, each medicine 21
It.When giving the processing of 17-AAG glucoside every time, the size of nude mice knurl, calculation formula are measured are as follows: knurl product V=1/2
(LxW2).After administration 21 days, nude mice is dislocated through vertebra and is put to death, knurl is separated out of nude mouse, is cleaned with PBS, filter paper
It dries, takes pictures and weigh the weight of tumor tissue.Statistical procedures: data are handled with SPSS16.0 statistical software, numerical value is with equal
Number ± standard deviation indicates that experimental group is handled compared with blank group with single factor test variance and Dunnette-t inspection is handled, and P value is less than
When 0.05, it is believed that difference has statistical significance between two groups of data.
(2) experimental result is as shown in figure 3,17-AAG glucoside processing group compared with blank control group, has significant suppression
The activity of tumour growth processed, inhibiting rate is 51.8% (P < 0.05) when being administered 21 days.In addition, the size and weight result of tumour is aobvious
Show, the tumor weight of administration group is significantly less than port lid control group (P < 0.05).
Claims (7)
1.17-AAG glucoside, which is characterized in that there is the structural formula as shown in formula (I):
2. the preparation method of 17-AAG glucoside described in claim 1, which is characterized in that specifically:
Tris-HCl, 1mmol/L MgCl for being 6.0~9.6 by pH value containing 25mmol/L2, 25mg YjiC, 3~6mmol/L
The reaction solution 50mL of UDP-glucose and 3mmol/L 17-AAG reacts 6~9 hours at 30 DEG C；After reaction, 100 DEG C of water
Bath heating 10 minutes, eliminates the activity of glycosyl transferase, then 50mL ethyl acetate is added into reaction solution and extracts to reaction solution
It takes, obtains ethyl acetate extract, through half preparation efficient liquid phase purification, obtain 17-AAG glucoside.
3. 17-AAG glucoside application in preparation of anti-tumor drugs described in claim 1.
4. 17-AAG glucoside application in preparation of anti-tumor drugs according to claim 3, it is characterised in that: institute
The 17-AAG glucoside stated is configured to for oral or injection administration.
5. 17-AAG glucoside application in preparation of anti-tumor drugs according to claim 3, it is characterised in that: institute
The tumour cell stated is the tumour cell of digestive system, gastrointestinal system and reproductive system position.
6. 17-AAG glucoside application in preparation of anti-tumor drugs according to claim 5, it is characterised in that: institute
The tumour cell stated is liver cancer cells, colon cancer cell and breast cancer cell.
7. 17-AAG glucoside application in preparation of anti-tumor drugs as claimed in claim 4, it is characterised in that: described
At pharmaceutical composition, the dosage form of pharmaceutical composition includes for 17-AAG glucoside or its salt and drug excipient or vehicle group
Liquid preparation, tablet, granule, electuary, capsule and pill, capsule, sustained release agent, pill or oral cavity disintegration preparation.
Priority Applications (1)
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| CN111138444A (en) * | 2020-01-08 | 2020-05-12 | 山东大学 | Epothilone B glucoside compounds and enzymatic preparation and application thereof |
| CN111205343A (en) * | 2020-01-08 | 2020-05-29 | 山东大学 | Nitrogen acetyl glucoside or galactoside compound of epothilone B, and enzymatic preparation and application thereof |
| CN112553231A (en) * | 2020-12-25 | 2021-03-26 | 华南农业大学 | Recombinant human heat shock protein HSP90-His and expression and purification method thereof |
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| CN111138444A (en) * | 2020-01-08 | 2020-05-12 | 山东大学 | Epothilone B glucoside compounds and enzymatic preparation and application thereof |
| CN111205343A (en) * | 2020-01-08 | 2020-05-29 | 山东大学 | Nitrogen acetyl glucoside or galactoside compound of epothilone B, and enzymatic preparation and application thereof |
| CN111205343B (en) * | 2020-01-08 | 2022-06-14 | 山东大学 | Nitrogen acetyl glucoside or galactoside compound of epothilone B, and enzymatic preparation and application thereof |
| CN112553231A (en) * | 2020-12-25 | 2021-03-26 | 华南农业大学 | Recombinant human heat shock protein HSP90-His and expression and purification method thereof |
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