CN100484951C - Panaxcoside secondary glucoside fatty acid ester compound, its preparation method and medicinal composition using said compound as active ingredient - Google Patents
Panaxcoside secondary glucoside fatty acid ester compound, its preparation method and medicinal composition using said compound as active ingredient Download PDFInfo
- Publication number
- CN100484951C CN100484951C CNB2004101554932A CN200410155493A CN100484951C CN 100484951 C CN100484951 C CN 100484951C CN B2004101554932 A CNB2004101554932 A CN B2004101554932A CN 200410155493 A CN200410155493 A CN 200410155493A CN 100484951 C CN100484951 C CN 100484951C
- Authority
- CN
- China
- Prior art keywords
- compound
- ethyl acetate
- straight chain
- acyl group
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
A secondary ginsenosides ester of fatty acid, its preparing process, its medicinal composition and the application of said compound and its medicinal composition in preparing medicine for treating cancer are disclosed.
Description
Technical field:
The present invention relates to ginsenoside secondary glycoside (being called for short M1) fatty acid ester compound (being called for short FM1), and disclose molecular structure, the synthetic method of this compound and take the pharmaceutical composition that this compound is activeconstituents, belong to the semi-synthetic technical field of Effective Component of Chinese Medicine.
Background technology:
Ginsenoside secondary glycoside involved in the present invention (being called for short M1) is the ginsenoside secondary glycoside that bacterium metabolism produces in intestines, and M1 can obtain by enzymolysis or fermentation process.Find after deliberation, M1 has very strong antitumour activity.M1 forms fatty acid ester (being called for short EM1) with lipid acid in liver, it than M1 the integration time in liver long.And find by the pharmacokinetic of M1, EM1, it is active that the M1 fatty acid ester that M1 fatty acid esterification forms (being called for short EM1) has improved its anti-tumor in vivo, and reduced cytotoxicity.Therefore, the antitumour activity of ginseng comes from oral ginseng by the metabolin M 1 of bacterium metabolism formation in intestines and the EM1 of intrahepatic fat acid estersization formation, and in intestines, bacterium metabolin M 1 can directly be brought into play antitumous effect.
Before I am engaged in this research, through retrieval, have no the bibliographical information of above-mentioned synthetic product.
Summary of the invention:
The object of the present invention is to provide a class to there is ginsenoside secondary glycoside fatty acid ester (the being called for short FM1) compound of pharmaceutical use.
Another object of the present invention has been to provide the synthetic and separation method of ginsenoside secondary glycoside fatty acid ester (being called for short FM1) compound, is suitable for industrialized mass.
Further aim of the present invention has been to provide a kind of pharmaceutical composition that is used for the treatment of cancer that this compound is activeconstituents of take.
Another object of the present invention is above-claimed cpd and the purposes of composition aspect the medicine of preparation treatment cancer.
The ester compound that the present invention use M1 and C8 ~ C16 fat acyl chloride are synthetic, through Pharmacological Activity Screening, effective for kinds cancer.
The compounds of this invention has following general structural formula (I):
Wherein: R is C
8~ C
16fatty acyl group, this compound is referred to as FM1.
Above-mentioned C
8~ C
16fatty acyl group is the acyl group with the straight or branched carboxylic acid of 8-16 carbon atom, and aliphatic carboxylic acid wherein comprises the saturated or unsaturated side chain lipid acid of naturally occurring saturated or unsaturated straight chain fatty acid and synthetic.The preferred compound of the present invention is that wherein R is C
8~ C
16the formula of straight chain fatty acyl group (I) compound; Preferably wherein R is C
8~ C
16the formula of saturated straight chain fatty acyl group (I) compound; Particularly preferably being wherein R is C
8~ C
16formula (I) compound of saturated straight chain even carbon fatty acyl group; Most preferably wherein R is C
16the formula of saturated straight chain fatty acyl group (I) compound.
Wherein R is C
16the formula of saturated straight chain fatty acyl group (I) compounds process for production thereof comprises the following steps:
A. the ginsenoside secondary glycoside of ginseng being prepared through microorganism fermentation or enzymolysis, by ODS reversed-phase column, or silica gel column chromatography is separated, with methanol solution wash-out;
B. collect and be rich in ginsenoside secondary glycoside elutriant part, reclaim methyl alcohol, be concentrated into dryly, obtain ginsenoside secondary glycoside, be called for short M1;
C. get M1 and be dissolved in ethyl acetate, in the situation that agitator stirs, add saturated sodium bicarbonate aqueous solution, under ice-water bath condition, add mole number 2-20 saturated straight chain palmitoyl chlorine doubly, under room temperature, stir and spend the night.Then with separating funnel, ethyl acetate layer is separated with water layer, and by ethyl acetate aqueous layer extracted repeatedly, ethyl acetate is merged, centrifugal, supernatant liquor water rinses repeatedly, ethyl acetate is reclaim under reduced pressure under 0-20 ℃ of cold condition, dissolve with methanol, filtration for product, and methanol solution concentrating under reduced pressure obtains dry-matter;
D. by gained dry-matter through HPLC, C-18 post, 100% methanol-eluted fractions, obtain 20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-Palmitic acid ester [20-O-β-D-glucopyranosyl (6-O-palmitoyl)-20 (S)-protopanaxadi], be called for short PM1; Yield is counted 35-60% with M1.
Gained monomeric compound can obtain the sterling of product of the present invention by re-crystallizing in ethyl acetate.
R is other C
8~ C
16the formula of fatty acyl group (I) compound, can, with the acyl chlorides of corresponding carbon atom, make according to above-mentioned preparation method.And inventor's discovery, adopt the inventive method, utilize C
8~ C
16the yield of the formula of fatty acyl group (I) compound, apparently higher than other yields of the formula of high-carbon fatty acyl group (I) compound more, if R is C
18the yield of the formula of saturated or unsaturated straight chain fatty acyl group (I) compound only has 5-20%.
The present invention's research shows that the fatty acid ester of M1 has obviously strengthened the antitumour activity of M1, especially FM1, particularly PM1.Pharmacokinetic by M1, PM1 finds that the formation of M1 fatty acid esterification FM1, particularly PM1 have improved its anti-tumor in vivo activity.Administration M1 or PM1 in mouse inoculation tumor cell of liver, M1 treatment group inhibiting rate is 23%, but not obvious with control group comparing difference; With the PM1 processing of same dose, obviously suppress tumor growth, remarkable with the poor heteropole of control group, also remarkable with M1 treatment group comparing difference.After M1 administration, optionally enter soon liver, in 10 minutes, reach climax, and from liver, discharge rapidly; And PM1 promptly accumulates in liver, although As time goes on PM1 also reduces gradually, the PM1 (the μ g/g of wet tissue) that surpasses dosage 25% can retain 24 hours in liver.This more effectively has identical reason with the cytosine arabinoside treatment hematologic malignancies after fatty acid esterification than cytosine arabinoside itself.Therefore, the enhancing of PM1 anti-tumor activity may the residence time in liver extends relevant with it.
Contriver is through also discovery of pharmacological experiment screening, although the compounds of this invention FM1 and R are C
18the formula of saturated or unsaturated straight chain fatty acyl group (I) compound (SM1 or OM1) all has anticancer physiologically active; various cancers are had to inhibition, alleviation and therapeutic action; but obviously the anticancer physiologically active of the compounds of this invention FM1 is stronger; better to the inhibition of various cancers, alleviation and result for the treatment of; the PM1 in FM1 particularly; the ability that anti-lung carcinoma cell shifts is obviously better than SM1, OM1, and toxicity is lower.
Pharmaceutical composition of the present invention contain treat effective dose above-mentioned general formula (I) compound F 17-hydroxy-corticosterone M1 for living
Property composition, and contain one or more pharmaceutically acceptable carriers.
Compound of the present invention and pharmaceutical compositions can be used for the medicine of the various cancers of preparation treatment.
Pharmaceutically acceptable carrier described above refers to the pharmaceutical carrier of pharmaceutical field routine, comprises that thinner, vehicle are as water etc.; Weighting agent is as starch, sucrose etc.; Tamanori is as Mierocrystalline cellulose and derivative, gelatin; Wetting agent is as glycerine; Disintegrating agent is as sodium bicarbonate, calcium carbonate etc.; Absorption enhancer is as quaternary ammonium salt; Tensio-active agent is as high-carbon fatty alcohol; Absorption carrier is as kaolin, soap clay; Lubricant is as talcum powder, calcium stearate and polyethylene glycol etc.; Can also in composition, add other assistant agent as sweeting agent, flavouring agent etc.
The compounds of this invention can be applied to cancer patients's treatment with the form of composition by oral, rectum, vein, muscle or parenteral admin mode.Also can be according to the conventional production method of pharmaceutical field, as make its activeconstituents and one or more carriers or medicament mixed, prepare various formulations as tablet, electuary, capsule, suppository, sprays etc., preferred form is tablet, electuary, capsule, suppository, sprays, sustained release dosage and injection; Particularly preferably be the preparation in the direct administration of lesions position.Pharmaceutical composition of the present invention contains the activeconstituents FM1 that weight ratio is 1%-99.5%, and preferably containing weight ratio is the activeconstituents FM1 of 45%-99.5%, and preferably containing weight ratio is the activeconstituents FM1 of 90%-99.5%.
Formulation rate of the present invention can be according to variations such as route of administration, patient age, body weight, disease type and severity, and per daily dose is 0.01-10mg/kg body weight, preferably 0.1-8mg/kg body weight.Can use by one or many.
Embodiment
The following examples can help the present invention of those skilled in the art comprehend, but do not limit the present invention in any way.
Embodiment 1.20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-Palmitic acid ester [20-O-β-D-glucopyranosyl (6-O-palmitoyl)-20 (S)-protopanaxadi] (being called for short PM1) preparation: the M1 by ginseng through glucolase fermentation preparation, by ODS reversed-phase column, or silica gel column chromatography is separated, after methanol solution wash-out, M1 elutriant part is rich in collection, reclaim methyl alcohol, be concentrated into dryly, obtain purity and be 98% M1; Get M150g and be dissolved in 1000ml ethyl acetate, in the situation that agitator stirs, add 1000ml water saturation sodium bicarbonate, under ice-water bath condition, add respectively saturated straight chain palmitoyl chlorine 420g, under room temperature, stir and spend the night.Then with separating funnel, ethyl acetate layer is separated with water layer, and by ethyl acetate aqueous layer extracted repeatedly, ethyl acetate is merged, centrifugal (3000rpm), supernatant liquor water rinses repeatedly, ethyl acetate is reclaim under reduced pressure under (20 ℃) condition at low temperatures, dissolve with methanol, filtration for product, and methanol solution concentrating under reduced pressure obtains dry-matter.Gained dry-matter, through HPLC, C-18 post, 100% methanol-eluted fractions, is obtained to 40g 20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-Palmitic acid ester (PM1), and yield counts 58.1% with M1.
Product is water white transparency oily thing, is dissolved in chloroform, methyl alcohol.
Liebermann---Burchard reaction is positive,
In TOF-MS spectrum, m/z[M+H]
+861.6, its molecular weight is 860.
1H-NMR(500MHz，py-d
5)δ：5.32(1H，t，J＝6.5Hz，H-24)，5.11(1H，d，J＝7.5Hz，glc-H-1)，
5.02(1H，d，J＝11.0Hz，glc-Ha-6)，4.63(1H，dd，J＝7.0，11.0Hz，glc-Hb-6)，4.18(1H，t，J＝8.5Hz，glc-H-4)，4.16(1H，t，J＝8.5Hz，glc-H-4)，3.98(1H，t，J＝9.5Hz，H-12α)，3.95(1H，t，J＝8.0Hz，glc-H-2)，3.92(1H，t，J＝9.0Hz，glc-H-5)，3.39(1H，dd，J＝5.0，11.0Hz，H-3α)，2.59(1H，m，H-17α)，2.55(1H，m，Hb-23)，2.43(1H，m，Hb-22)，2.31(1H，m，Ha-23)，2.02(1H，m，Hb-11)，2.00(1H，m，H-13)，1.88(1H，m，Hb-2)，1.85(1H，m，Hb-16)，1.82(1H，m，Ha-22)，1.80(1H，m，Ha-2)，
1.69 (1H, m, Hb-1), 1.67 (3H, s, Me α), 1.64 (3H, s, Me-26), 1.62 (3H, s, Me-27), 1.59 (1H, m, Hb-6), 1.55 (1H, m, Ha-1), 1.53 (1H, m, Hb-15), 1.50 (1H, m, Hb-7), 1.45 (1H, m, Ha-6), 1.42 (1H, m, H-9 α), 1.39 (1H, Ha-10), 1.31 (1H, Ha-7), 1.20 (3H, s, Me-28 α), 1.03 (1H, m, Ha-15), 1.02 (3H, s, Me-29 β), 0.98 (3H, s, 19 β), 0.94 (3H, s, Me-30 α), 0.89 (1H, m, Ha-1), 0.87 (3H, s, Me-18 β) 0.85 (3H, t, J=7.5Hz, lipid acid terminal M e), 0.79 (1H, d, J=11.0Hz, H-5).
13c-NMR (125MHz, py-d
5) δ: its ownership refers to table 1, its carbon ownership and M1 comparison,
13on C-NMR wave spectrum, the C of glucosyl residue
6signal by δ 62.7 to low field displacement to δ 64.7 (△ 2.0); ?
1on H-NMR wave spectrum, glucosyl residue-CH
2oH signal is by δ 4.44 (1H, d, J=11.0Hz, glc-Ha-6) and 4.27 (1H, m, glc-Hb-6) respectively to low field displacement to δ 5.02 (1H, d, J=11.0Hz, glc-Ha-6) and 4.63 (1H, d, J=7.0,11.0Hz, glc-Hb-6).And C3 signal (δ 78.0) and H-3 signal α [δ 3.39 (1H, dd, J=5.0,11.0Hz)] are in full accord.Therefore, the CH of fatty acid residue and glucosyl residue
2oH dehydrating condensation.Because the molecular weight of M1 is 622, the molecular weight of Palmitic acid is 256 (C
16h
32o
2), both condensation product molecular weight are 860, this is identical with the institute molecular weight of surveying.Determine that this compound is M1 Palmitic acid acid esters, i.e. 20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-cetylate, referred to as PM1.
Table 1. compound PM1 and M1's
13c-NMR data
The preparation of embodiment 2.20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-lauric acid ester (being called for short DM1): as embodiment 1 preparation M1; Get M150g and be dissolved in 1000ml ethyl acetate, in the situation that agitator stirs, add 1000ml water saturation sodium bicarbonate, under ice-water bath condition, add respectively saturated straight chain 12 carbonyl chloride 70g, under room temperature, stir and spend the night.Then with separating funnel, ethyl acetate layer is separated with water layer, and by ethyl acetate aqueous layer extracted repeatedly, ethyl acetate is merged to centrifugal (3000rpm), supernatant liquor
Water rinses repeatedly, and ethyl acetate is reclaim under reduced pressure under (20 ℃) condition at low temperatures, dissolve with methanol, filtration for product, and methanol solution concentrating under reduced pressure obtains dry-matter.Gained dry-matter, through HPLC, C-18 post, 100% methanol-eluted fractions, is obtained to 23g 20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-lauric acid ester (DM1), and yield counts 35.7% with M1.Product is water white transparency oily thing, is dissolved in chloroform, methyl alcohol.
The preparation of embodiment 3.20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-octanoate (being called for short OM1): as embodiment 1 preparation M1; Get M150g and be dissolved in 1000ml ethyl acetate, in the situation that agitator stirs, add 1000ml water saturation sodium bicarbonate, under ice-water bath condition, add respectively saturated straight chain capryl(yl)chloride 156g, under room temperature, stir and spend the night.Then with separating funnel, ethyl acetate layer is separated with water layer, and by ethyl acetate aqueous layer extracted repeatedly, ethyl acetate is merged, centrifugal (3000rpm), supernatant liquor water rinses repeatedly, ethyl acetate is reclaim under reduced pressure under (20 ℃) condition at low temperatures, dissolve with methanol, filtration for product, and methanol solution concentrating under reduced pressure obtains dry-matter.Gained dry-matter, through HPLC, C-18 post, 100% methanol-eluted fractions, is obtained to 34.7g20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-octanoate (OM1), and yield counts 58% with M1.Product is water white transparency oily thing, is dissolved in chloroform, methyl alcohol.
Following pharmacological evaluation has confirmed the Pycnorus cinnabarius of the compounds of this invention.
The restraining effect of experimental example 4. Palmitic acid esters (PM1) to liver tumor, gastric tumor growth
1. experiment material
Pharmaceutical chemicals and test sample: PM1: synthetic by this experiment, purity 97%, tween: purchased from Hualian Pharmaceutical Co., Ltd., Shanghai, lot number 020803, endoxan: purchased from China Medicine (Group) Shanghai Chemical Reagent Co.,, lot number 20001110.
Laboratory animal: ICR mouse, ♀ ♂ half and half, body weight 18 preclinical medicine institute of～22g，You Jilin University Experimental Animal Center provide.Experimental animal feeding is that in the plastics cage of iron wire fence, bottom is covered with sawdust at top, and black and white circulation in 12 hours, supplies food and water at any time.
2. experimental technique
Adopt animal-transplanted tumor model, with physiological saline group, positive drug endoxan group, compare, PM1 antitumour activity is carried out to screening and assessment, as follows with concrete operations:
The restraining effect of A.PM1 to rat liver cancer ascitic type (HepA) Growth of Cells: the oncocyte going down to posterity in mouse peritoneal 7 days (HepA) is taken out under aseptic condition, with physiological saline washing 2 times, with normal saline dilution, oncocyte number in every milliliter counts, adjust cell concn to 1? 07ml-1, make abdominal injection, every the right armpit subcutaneous vaccination of mouse 0.2ml knurl liquid to acceptor mouse.Mouse surviving rate 100%, similar on host's impact, interindividual variation is very little.Choose 30 of mouse, be divided at random 3 groups, (injecting normal saline group, positive drug endoxan group, PM1 administration group), next day intraperitoneal administration, positive drug endoxan group mouse dosage 30mg/kg every other day administration, PM1 administration group mouse dosage 20mg/kg/day.Continuous 10 days, after drug withdrawal, 24 hours de-necks were put to death mouse, separate tumour electronic scale and weigh, and calculate tumor suppression percentage.
The restraining effect of B.PM1 to Mouse Gastric Cancer (MFC) Growth of Cells: the oncocyte going down to posterity in mouse peritoneal 7 days (MFC) is taken out under aseptic condition, with physiological saline washing 2 times, with normal saline dilution, oncocyte number in every milliliter counts, adjust cell concn to 1 * 107ml-1, make abdominal injection, every the right armpit subcutaneous vaccination of mouse 0.2ml knurl liquid to acceptor mouse.Mouse surviving rate 100%, similar on host's impact, interindividual variation is very little.Choose 30 of mouse, be divided at random 3 groups, (injecting normal saline group, positive drug endoxan group, PM1 administration group), next day intraperitoneal administration, positive drug endoxan group mouse dosage 30mg/kg every other day administration, PM1 administration group mouse dosage 20mg/kg/day.Continuous 10 days, after drug withdrawal, 24 hours de-necks were put to death mouse, separate tumour electronic scale and weigh, and calculate tumor suppression percentage.
3. the statistical study of experimental data
The statistics of experimental data adopts t-method of inspection; Result with mean add and subtract standard deviation (
± S) represent.
Tumor control rate (%)=(1-T/C) * 100%
T: the heavy C of the average knurl of experimental group: the average knurl weight of control group
4. experimental result and conclusion
The restraining effect of A.PM1 to rat liver cancer (HepA) Growth of Cells
PM1 administration group knurl is heavily starkly lower than the control group that physiological saline is processed, with the poor heteropole of control group remarkable (p<0.001), illustrate that PM1 has significantly suppressed the growth of liver cancer cell in Mice Body, PM1 administration group and positive drug endoxan group relatively act on not as good as endoxan, PM1 administration group tumor control rate is 53.66%, and positive drug endoxan group tumor control rate is 73.15%.As table 2.
The impact of table 2. different treatment on mouse tumor cell (HepA)
| Process group | Number of animals | Tumour heavy (g) | Inhibiting rate (%) |
| Contrast | 10 | 2.97±0.54 | —— |
| CTX?30mg/kg | 10 | 0.86±0.35*** | 73.15 |
| PM120mg/kg | 10 | 1.44±0.57*** | 53.66 |
In * * P<0.001 table, data are 3 test averages.
The restraining effect of B.PM1 to Mouse Gastric Cancer (MFC) growth
PM1 administration group knurl is heavily starkly lower than the control group that physiological saline is processed, with control group comparing difference remarkable (p<0.05), illustrate that PM1 has significantly suppressed the growth of stomach cancer cell in Mice Body, PM1 administration group and positive drug endoxan group relatively act on not as good as endoxan, PM1 administration group tumor control rate is 54%, and positive drug endoxan group tumor control rate is 72.11%.As table 3.
The impact of table 3. different treatment on Mouse Gastric Cancer cell (MFC) growth
| Process group | Number of animals | Tumour heavy (g) | Inhibiting rate (%) |
| Contrast | 10 | 3.32±1.39 | —— |
| CTX30mg/kg | 10 | 0.99±0.51*** | 72.11 |
| PM120mg/kg | 10 | 1.60±0.68** | 54.00 |
In * P<0.01 * * * P<0.001 table, data are 3 test averages.
The restraining effect of experimental example 5.PM1 to melanoma growth
1. liver transplantation melanoma
Laboratory animal: ICR mouse, ♀ ♂ half and half, body weight 18～22g, in age in 12-16 week, chooses 30 of mouse, is divided at random 3 groups, and black and white circulation in 12 hours, supplies food and water at any time.
2. experimental technique
Mouse peritoneal is injected to Sodital (50mg/kg) anesthesia, with injection into liver inoculation B16F-10 tumour cell (2 x10
5individual cell) in contrast; In Mouse Liver, inject B16F-10 tumour cell (2 x 10 simultaneously
5individual cell) and M1 (5mg/kg); In Mouse Liver, inject B16F-10 tumour cell (2 x 10 simultaneously
5individual cell) and PM1 (5mg/kg) respectively as processing.After 10 days, slaughter mouse and extract liver, weigh tumor weight.
3. the statistical procedures of data
The statistics of experimental data adopts t-method of inspection; Result with mean add and subtract standard deviation (
± S) represent.
4. experimental result and conclusion
Calculate the mean value (EM ± SD) of 10 mouse tumor weights, Epidemiological Analysis by statistics, result is as table 4.
Result of study shows, after M1 forms fatty acid ester PM1, antitumour activity obviously strengthens.
The restraining effect of table 4.PM1 to melanoma growth
| Process group | Number of animals | Tumour heavy (g) | Inhibiting rate (%) |
| Contrast | 10 | 2.4±0.56 | —— |
| M15mg/kg | 10 | 1.85±0.47* | 22.9 |
| PM15mg/kg | 10 | 0.95±0.35# | 60.4 |
#, p<0.002 with compare; *, p<0.02 and M1 comparison.
The splenic lymphocyte that experimental example 6PM1 processes is to B16-F10 cytotoxic effect
1. experiment material and method
Lymphocytic preparation: liver or spleen are suspended in blood cytolysate [0.17MNH by stainless steel mesh
4cl, 0.01mM EDTA, 0.1M Tris (pH7.3)] in, centrifugal 6 minutes of 100xg room temperature.Cell granulations is washed 2 times with full medium, and lymphocyte and 20 μ M 2 mercapto ethanols that liver is cultivated are added in full medium.
2. cancer cells toxic action is measured
Cancer cells (1 x 10
6/ hole) with liver or splenic lymphocyte (1-3 x 10
6/ hole) in full medium, cultivate, and under PM1 (0.1-10 μ M) existence or non-existent condition, 5% CO
2in gas, 37 ℃ of cultivations.After 3 days, with trypsinase, collect AC and examine under a microscope and measure cancer cells number.Another group splenic lymphocyte (1 x 10
6) use in advance PM1 (0-240 μ M) to process.After 2 days, splenic lymphocyte be divided into AC and non-AC and immediately respectively with B16-F10 cell (2 x 10
4) cultivate.Parallel laboratory test: cancer cells and PM1 single culture.After 1 day, with trypsinase, collect AC and measure cancer cells number.The ability of lymphocytolysis cancer cells is applied following formula and is calculated: solvency power (%)=(1-test group/control group) x 100, wherein test group=cancer cells with lymphocyte PM1 exist or non-existent condition under, the cancer cells number of cultivation; The cancer cells number that control group=do not have lymphocyte is cultivated.
By the PM1 pre-treatment splenic lymphocyte of 2 days, be divided into two groups: AC and non-AC and respectively with B16-F10 cell cultures.Parallel laboratory test: cancer cells and PM1 single culture.After 1 day, with trypsinase, obtain AC and measure cancer cells number.
3. the statistical procedures of data
The statistics of experimental data adopts t-method of inspection; Result with mean add and subtract standard deviation (
± S) represent.
4. experimental result and conclusion
Measure the cancer cells number under PM1 different concns, get the growth rate that lower 3 the dish mean numbers of each concentration of PM1 (EM ± SD) calculate cancer cells.Result is as table 5.
The splenic lymphocyte that table 5.PM1 processes is to B16-F10 cytotoxic effect
| PM1 concentration (μ m) is processed group | 0 | 60 | 120 | 240 |
| Cancer cells+PM1 | 100 | 100 | 100 | 100 |
| Cancer cells+AC+PM1 | 100 | 100 | 100 | 100 |
| Cancer cells+non-AC+PM1 | 100 | 85 | 60 | 30 |
Result shows: cancer cells is cultivated with the non-AC of processing with PM1 in advance, and with the variation of seeing concentration, growth of cancer cells inhibiting rate increases.And AC does not have restraining effect to the growth of cancer cells.Also not effect of M1 separately.These results show the restraining effect of PM1 to growth of cancer cells and transfer, and possible right and wrong AC is to the stimulation of the cancer cells state destroying and activate relevant.
The comparative studies of embodiment 7.DM1, PM1, SM1, OM1 anti-tumor activity
1. experiment material
Lewis lung tumor cell
Laboratory animal: ICR mouse, ♀ ♂ half and half, body weight 19～23g, in age in 12-16 week, chooses 50 of mouse, is divided at random 5 groups, and black and white circulation in 12 hours, supplies food and water at any time.
2. experimental technique
Mouse peritoneal is injected to Sodital (50mg/kg) anesthesia, and after intravenous inoculation cancer cells, with the dosage of 3mg/Kg, respectively to different treatment group oral administration, control group is given corresponding suspensoid, after 10 days, slaughters mouse and takes out lungs and weigh.
3. the statistical procedures of data
Statistics is as table 6.The statistics of experimental data adopts t-method of inspection; Result with (
± SD) represent.
Table 6.DM1, PM1, SM1, OM1 oral administration are to the spontaneous useless curative effect shifting of Lewis lung cancer
| Group | Dosage (mg/kg) | Dosage regimen | Number of animals (begin/eventually) | The weight of animals (g, begin/eventually) |  |
Inhibiting rate (%) |
| DM1 | PM1 | SM1 | OM1 contrast | 3.00 3.00 3.00 3.00 corresponding suspensions | po×10qod po×10qod po×10qod po×10qod po×10qod | 10/10 10/10 10/10 10/10 10/10 | 19.1±21.6 19.2±21.7 19.5±21.4 19.0±22.3 19.3±22.7 | 1.67±0.23 1.70±0.23 4.60±5.17 4.30±5.10 7.60±6.60 | 77.48** 77.63** 39.47** 43.42** |
* P<0.05 * * P<0.01, in table, data are 3 experiment averages.
4. experimental result
DM1, PM1, SM1, OM1 oral administration is to the spontaneous useless curative effect shifting of Lewis lung cancer, DM1, PM1, SM1, OM1 oral administration group knurl is heavily starkly lower than control treatment group, with the poor heteropole of control group remarkable (p<0.01), DM1 is described, PM1, SM1, OM1 has significantly suppressed the growth of lung carcinoma cell in Mice Body to be shifted, DM1, PM1 administration group and SM1, OM1 administration group relatively acts on and is obviously better than the latter, DM1, PM1 administration group tumor control rate is respectively 77.48%, 77.63%, and SM1 administration group tumor control rate is 39.42%, OM1 administration group tumor control rate is 43.42%.Result shows: the ability that the anti-lung carcinoma cell of DM1, PM1 shifts is obviously better than SM1, OM1.
The preparation of embodiment 8. anticancer tablets: get the compound 10g of embodiment 1 preparation, add the medicinal cyclodextrin 30g of vehicle, mix, granulation compressing tablet, makes tablet, every containing PM1 10mg.
The preparation of embodiment 9. anti-cancer capsules: get the compound 10g of embodiment 1 preparation, add the medicinal cyclodextrin 10g of vehicle, mix, granulation, incapsulates, every containing PM1 10mg.
The preparation of embodiment 10. anticancer injections: get the compound 10g of embodiment 3 preparations, add Proplyleng Glycol 200ml, water for injection 800ml, dissolves membrane filtration; Membrane filtration (0.2 μ m), packing, every bottle of 2ml, containing PM120mg.All operations all should carry out under aseptic condition.
The preparation of embodiment 11. anti-cancer capsules: get the compound 10g of embodiment 2 preparations, add the medicinal cyclodextrin 10g of vehicle, mix, granulation, incapsulates, every containing PM1 10mg.
Claims (8)
- [claim 1]The panaxcoside secondary glucoside fatty acid ester compound with antitumous effect, is characterized by and have following structure:
Wherein R is C8 ~ C16 fatty acyl group. - [claim 2]According to the compound of claim 1, it is characterized by R is C8 ~ C16 straight chain fatty acyl group.
- [claim 3]According to the compound of claim 2, it is characterized by R is C8 ~ C16 saturated straight chain fatty acyl group.
- [claim 4]According to the compound of claim 3, it is characterized by C8 ~ C16 saturated straight chain even carbon fatty acyl group.
- [claim 5]According to the compound of claim 4, it is characterized by R is C16 saturated straight chain fatty acyl group.
- [claim 6]The preparation method of the compound of claim 5, is characterized by and comprise the following steps:A. the ginsenoside secondary glycoside of ginseng being prepared through microorganism fermentation or enzymolysis, by ODS reversed-phase column, or silica gel column chromatography is separated, with methanol solution wash-out;B. collect and be rich in ginsenoside secondary glycoside elutriant part, reclaim methyl alcohol, be concentrated into dryly, obtain ginsenoside secondary glycoside, be called for short M1;C. get M1 and be dissolved in ethyl acetate, in the situation that agitator stirs, add saturated sodium bicarbonate aqueous solution, under ice-water bath condition, add the saturated straight chain palmitoyl chlorine of mole number 2-20 multiple, under room temperature, stir and spend the night; Then with separating funnel, ethyl acetate layer is separated with water layer, and by ethyl acetate aqueous layer extracted repeatedly, ethyl acetate is merged, centrifugal, supernatant liquor water rinses repeatedly, ethyl acetate is reclaim under reduced pressure under 0-20 ℃ of cold condition, dissolve with methanol, filtration for product, and methanol solution concentrating under reduced pressure obtains dry-matter;D. by gained dry-matter through HPLC, C-18 post, methanol-eluted fractions, obtain 20-(S)-protopanoxadiol-20-O-β-D-Glucopyranose-6-O-Palmitic acid ester, be called for short PM1.
- [claim 7]In claim 1-5, the compound of any one is applied in the medicine of preparation treatment cancer.
- [claim 8]The pharmaceutical composition that is used for the treatment of cancer, is characterized by the compound and the pharmaceutically acceptable carrier that wherein contain the claim 1 for the treatment of significant quantity.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004101554932A CN100484951C (en) | 2004-12-01 | 2004-12-01 | Panaxcoside secondary glucoside fatty acid ester compound, its preparation method and medicinal composition using said compound as active ingredient |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004101554932A CN100484951C (en) | 2004-12-01 | 2004-12-01 | Panaxcoside secondary glucoside fatty acid ester compound, its preparation method and medicinal composition using said compound as active ingredient |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1651452A CN1651452A (en) | 2005-08-10 |
| CN100484951C true CN100484951C (en) | 2009-05-06 |
Family
ID=34869731
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004101554932A Expired - Fee Related CN100484951C (en) | 2004-12-01 | 2004-12-01 | Panaxcoside secondary glucoside fatty acid ester compound, its preparation method and medicinal composition using said compound as active ingredient |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100484951C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI717385B (en) * | 2015-09-30 | 2021-02-01 | 南韓商愛茉莉太平洋股份有限公司 | Ginsenoside fatty acid ester compounds, preparation method thereof, and cosmetic composition comprising the same |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101921305B (en) * | 2010-06-23 | 2012-09-05 | 大连大学 | Method for preparing ginsenoside metabolite M1 fatty acid monoester compound |
| CN102827233B (en) * | 2011-06-14 | 2016-04-13 | 李晓辉 | Ginsenoside Compound K ester derivative and the application in the medicine of preparation control arteriosclerosis thereof |
| CN102942610A (en) * | 2012-11-26 | 2013-02-27 | 吉林农业大学 | Preparation of ginsenoside M1 n-butyrate and application of antidiabetic medicines of ginsenoside M1 n-butyrate |
| CN107619847A (en) * | 2017-11-09 | 2018-01-23 | 宁波润沃生物科技有限公司 | A kind of method that biofermentation prepares panaxoside metabolin |
-
2004
- 2004-12-01 CN CNB2004101554932A patent/CN100484951C/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| Oleoyl triterpene. Hideo hasegawa,et al.Journal of traditional medicines,Vol.17 No.5. 2000 |
| Oleoyl triterpene. Hideo hasegawa,et al.Journal of traditional medicines,Vol.17 No.5. 2000 * |
| pharmcokinetics of ginsenoside. Hideo hasegawa,et al.bio. pharm. bull.,Vol.23 No.3. 2000 |
| pharmcokinetics of ginsenoside. Hideo hasegawa,et al.bio. pharm. bull.,Vol.23 No.3. 2000 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI717385B (en) * | 2015-09-30 | 2021-02-01 | 南韓商愛茉莉太平洋股份有限公司 | Ginsenoside fatty acid ester compounds, preparation method thereof, and cosmetic composition comprising the same |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1651452A (en) | 2005-08-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101224215B (en) | Uses of ursolic acid saponin and oleanolic acid saponin in preparing medicine for increasing white blood cell and/or blood platelet | |
| CN102344481A (en) | Derivatives of 3-O-caffeoyloleanane type pentacyclic triterpene, preparation method thereof and application thereof | |
| CN100484951C (en) | Panaxcoside secondary glucoside fatty acid ester compound, its preparation method and medicinal composition using said compound as active ingredient | |
| JP2017518381A (en) | Medicinal use of anti-tumor for rutile pentacyclic triterpene saponins compounds | |
| CN110437247B (en) | Mixed source terpenoid with liver protection function and application thereof | |
| CN102525929A (en) | Wogonin liposome preparation modified with glycyrrhetinic acid and preparation method thereof | |
| CN111333694B (en) | Application of hederagenin derivative in medicine for resisting myocardial anoxia reoxygenation injury | |
| CZ291612B6 (en) | Ara-C derivatives, pharmaceutical compositions in which the derivatives are comprised as well as their use | |
| CN102603847A (en) | Preparation method and medicinal application of ginsenoside Rh2 aliphatic ester compound | |
| CN100486595C (en) | Brazil mushroom soluble small molecular extract, its preparation technology and use | |
| US8759549B2 (en) | Compound extracted from Antrodia cinnamomea and pharmaceutical composition comprising the same | |
| CN101824065B (en) | Fatty-acid monoester compound of ginsenoside secondary glycoside Rh1 and preparation method thereof | |
| CN106631957A (en) | Antitumor compound targeting FAP-alpha enzyme and preparation method and application thereof | |
| CN106748939A (en) | The new bromine phenol thiosemicarbazide compound of one class and its preparation and medicine and purposes | |
| CN1300160C (en) | Fatty acid ester compounds of ginsenoside secondary glycoside compound K and method for preparing same | |
| CN105859823A (en) | Application of ilicis routundae cortex acid ester derivatives in preparation of anti-tumor drugs | |
| CN100381459C (en) | Adriamycin derivative and its preparing method and use | |
| CN107200769B (en) | A kind of rotundic acid derivative with prevention and treatment metastasis effect | |
| CN102250197B (en) | Preparation method and application of total steroidal saponin extracts of dwarf lilyturf roots | |
| CN114642688A (en) | Application of ganoderma lucidum spore oil in preparing medicine for prolonging life of tumor patient | |
| CN101293904B (en) | New compound, preparation method and application thereof | |
| KR100564383B1 (en) | Process for preparing ginsenoside derivatives | |
| CN102503897B (en) | 5-fluorouracil iodized oil derivative as well as preparation method and application thereof | |
| KR100547253B1 (en) | Treating and prevention of cancer with new ginsenoside derivatives | |
| WO2020063262A1 (en) | Application of 3'-deoxyinosine in preparation of drug, food or health product for multiple disease |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090506 Termination date: 20191201 |