CN101481394B - 2,3,4,6-tetra-O-acetyl-D-glucopyranosyl-[N,N'-bis(2-chloroethyl)]- phosphorodiamidate, and preparation and use thereof - Google Patents
2,3,4,6-tetra-O-acetyl-D-glucopyranosyl-[N,N'-bis(2-chloroethyl)]- phosphorodiamidate, and preparation and use thereof Download PDFInfo
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Abstract
The invention discloses 2,3,4,6-tetra-O-acetyl-D-glucopyranosyl-[N,N'-di(2-chloroethane)]-diamidophosphoric acid I, an alpha configuration II and a beta configuration III thereof. Anti-cancer pharmacodynamic tests prove that the compounds have inhibitory effect on mouse transplantation tumor, and a drug toxicology test results of drugs thereof prove that the drugs have low toxic and side effects and can be used for preparing anti-cancer drugs. The invention further provides a preparation method of compound I, II and III. The preparation method has few reaction steps, high yield and low cost, and is suitable for industrialized production.
Description
Technical field:
The invention belongs to the pharmaceutical chemistry technical field.Be specifically related to 2,3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides and its production and use.
Background technology:
The high incidence of cancer has been one of the factor of human health that constitutes a serious threat at present.Although people are developing the gene therapy method for the treatment of cancer gene engineering medicine and cancer, large-scale application in clinical, distance is also quite remote.Therefore in cancer therapy, need the little and eutherapeutic chemotherapeutic of the more toxic side effect of exploitation.Mustargen, podophyllotoxin and di(2-ethylhexyl)phosphate acid amides etc. all belong to mammiferous cytotoxin, but have because toxicity is too large, what have is difficult to enter cancer cells, thereby affects the treatment.In order to make the correlation function gene be easy to enter cancer cells, reduce toxic side effect, pharmacy men are combined cytotoxin usually with carbohydrate, carrier by delivery carbohydrate on the cytolemma is introduced cancer cells with anticarcinogen, then, under intracellular enzyme and the effect of enzyme system, discharge antioncogene, the anticancer growth.Therefore synthesized many cytotoxic carbohydrate derivates recent decades, first application carbohydrate of Vargha et al (1957) has synthesized nitrogen mustard derivatives as bio-carrier; Thomas (1961) has synthesized two (β-chloroethyl)-α of ethyl-N--D-6-galactopyranose methane amide ester with semi-lactosi, and mouse subcutaneous injection every day 2mg totally 14 days, reaches 40% to Mammals body of gland cancer inhibiting rate; Kitty et al. (1962) synthetic 2,3,4,6-four-O-ethanoyl-D-glucopyranosyl-[N, N '-two-β-chloroethyl] di(2-ethylhexyl)phosphate acid amides syrup, mouse oral 250mg.kg-1 every day, totally 7 days, Mammals body of gland inhibiting rate is reached 40～50%; Gudkova et al. (1968) synthesizes 2,3,5,6-, two-O-isopropylidene-D-mannopyranose base-1-isobutyl-phosphoric acid ester.Ovrutskii et al (1974) has also synthesized N-alkyl-N, 2,3,4 of N '-two (2-chloroethyl) diamide phosphoric acid, 6-four-O-glucopyranosyl ester.Nineteen eighty-three drugs approved by FDA with β-D-Glucopyranose, trade(brand)name " Vepesid " (etoposide, etoposide, anticarcinogen VP-16-213) that podophyllotoxin and formaldehyde are synthetic.Dickes et al. (1988) has synthesized β-D-glucopyranosyl [N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (Glufosfamide, Glc-IPM, GLU-IPM), in external use 4 * 10
-5During molar dose, the restraining effect of mouse honeycomb sarcoma growth is approximately 20% in two hours, and the cancers such as pancreas, mammary gland and leukemia are all had good curative effect.But from the synthetic Glufosfamide of glucose feed, reactions steps is too many, and yield is very low, and cost is high, and Glufosfamide extremely difficult crystallization in solvent, needs preparation dried frozen aquatic products or freeze-dried preparation.And Glufosfamide is polar material, and entering needs the transhipment by acceptor SAATI behind the human body, needs consumed energy.Chemically with on the zymetology also there is relative instability in it simultaneously, at the water more than 18 ℃, automatically degradable in the solution such as methyl alcohol and ethanol, may cause the different phosphoamide diffusion of cell toxicant, cause toxic side effect to increase and curative effect reduction (U.S.pat.NO.20040029815).Therefore, pharmaceutically Glufosfamide remains the difficulty that is being difficult to overcome.
Summary of the invention:
Technical problem to be solved by this invention is to overcome above-mentioned weak point, research and design good effect, the little anticarcinogen of side effect.
The invention provides the new compound 2,3,4 of a kind of formula I, 6-four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides.
AC=CH3CO-in the formula (I)
The compounds of this invention has α configuration (formula II) and beta comfiguration (formula III), and both are white powder, the fusing point of formula II: heat heat is analyzed as unimodal, and mp91.2 ℃, specific rotation: [α]
D 20-4.820 (C1, ethanol); The fusing point of formula III: the unimodal mp119.70 of differential thermal analysis ℃, specific rotation [α]
D 20-6.31 ° (C0.4, ethanol).
2,3,4; 6-four-O-ethanoyl-α-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (II) and 2,3; 4,6-, four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (III).
AC=CH3CO-among formula (II), (III).
Another object of the present invention has provided the preparation method of 2,3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (I).
The inventive method is first synthetic 2,3,4,6-, four-O-ethanoyl-α-D-Glucopyranose bromine and N take commercially available glucose and 2-chloroethyl amine hydrochloride as basic raw material, N '-two (2-chloroethyl)-two kinds of intermediates of di(2-ethylhexyl)phosphate acid amides.Above-mentioned two kinds of intermediates have been condensed into 2,3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (I).Synthetic product divides α-type (II) and two kinds of structures of β-type (III), obtains high purity α-type (II) and two kinds of products of β-type (III) through separation and purification and crystallization.
The inventive method specifically comprises the following steps:
(1) 2, the preparation of 3,4,6-, four-O-ethanoyl-α-D-Glucopyranose bromine
With glucose, aceticanhydride and phosphorus tribromide or bromine water and red phosphorus are raw material, and preparation 2,3,4,6 four-O-ethanoyl-α-D-Glucopyranose bromine reacts as follows:
(VI) is the D-Glucopyranose in the formula
(V) be 2,3,4,6-, four-O-ethanoyl-α-D-Glucopyranose bromine
Reaction (5 ℃～-10 ℃) under ice bath adds perchloric acid and makes catalyzer (1: 0.01～1: 0.3), and reaction temperature is no more than 40 ℃ in adding the glucose process.Crystallized product through silica gel G thin-layer chromatography and paper chromatography (propyl carbinol: acetic acid: water=4: 1: 5, V/V) and 105 ℃ of bakings automatically show afterwards brown spot.Crystallized product ethanolic soln and the straight fire of 3,5-edlefsen's reagent add the reaction of thermogenesis reddish-brown.
(2) preparation of [N, N '-two (2-chloroethyl)-di(2-ethylhexyl)phosphate acid amides]
Take phosphorus oxychloride and 2-chloroethyl amine hydrochloride as main raw material, preparation [its reaction is as follows for N, N '-two (2-chloroethyl)-di(2-ethylhexyl)phosphate acid amides (ifosfamide):
(VI) is [N, N '-two (2-chloroethyl)-di(2-ethylhexyl)phosphate acid amides] in the formula
Et
3N is that triethylamine, THF are tetrahydrofuran (THF)
This reaction is carried out in anhydrous solvent chloroform, methylene dichloride or the ethylene dichloride below-10 ℃, and triethylamine is used for the hydrochloric acid that neutralization generates.Make crystallized product VI; (propyl carbinol: acetic acid: water=4: 1: 5, V/V), phosphorus reagent (3mol/L H is decided in spray after the oven dry through silica gel G reagent thin-layer chromatography in the product crystallization
2SO
4: water: 2.5% ammonium molybdate: 10% xitix=1: 2: 1: 1, V/V) be blue spot.The 1%NaOH solution of product crystallization with decide the phosphorus reagent mix in 40～50 ℃ of insulations 30 minutes, solution is blue and reacts.
(3) 2, the preparation of 3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides
In the anhydrous solvents such as methylene dichloride or chloroform; 2; 3,4,6-, four-O-ethanoyl-D-Glucopyranose bromine (V) and [N; N '-two (2-chloroethyl)]-two kinds of intermediates of di(2-ethylhexyl)phosphate acid amides (VI) are condensed into 2; 3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N; N '-two (2-chloroethyl)]-the di(2-ethylhexyl)phosphate acid amides, its reaction is:
This reaction is that 30～70 ℃ are stirred or back flow reaction 20～25 hours.(propyl carbinol: acetic acid: water=4: 1: 5, V/V), decide phosphorus reagent and be blue spot by the rear spray of oven dry through the silica gel G thin-layer chromatography for product.The ethanolic soln of product (I) and the straight fire of 3,5-edlefsen's reagent add thermogenesis reddish-brown reaction, and decide the phosphorus reagent mix in 40～50 ℃ of insulations 30 minutes, are blue reaction.
(4) the α-type II of separating step (3) condensation product I and β type III:
Step (3) condensation product (I) is the mixture of α-type (II) and two kinds of configurations of β-type (III), through silica gel column chromatography (methylene dichloride: methyl alcohol=80: 20, V/V) separate, again by crystallization and recrystallization, then isolate 3～5% α-type (II) and 95% above β-type (III), and be used for acute toxicity test and the test of antitumor drug effect.
In the method for the invention, described step (3) condensation reaction 2,3,4,6-four-O-ethanoyl-α-D-Glucopyranose bromine is 1～1.5: 1 with the mol ratio of [N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides.Temperature of reaction is 30～70 ℃; PH is 6～9; The stirring reaction time is 16～30 hours; Used anhydrous solvent is chloroform, methylene dichloride, 1,2-ethylene dichloride, 1,1-ethylene dichloride, vinyl acetic monomer, tetrahydrofuran (THF), ether, benzene, toluene, sherwood oil or pyridine.Described step (4) silica gel column chromatography eluent is chloroform, methylene dichloride, ethylene dichloride, sherwood oil, tetrahydrofuran (THF), toluene, benzene, ether or vinyl acetic monomer or their mixed solvent; The crystallization of formula I, formula II or formula III compound and recrystallization solvent are chloroform, methylene dichloride, ethylene dichloride, vinyl acetic monomer, toluene, benzene, tetrahydrofuran (THF), sherwood oil, ether or pyridine or their mixed solvent, and the temperature of crystallization and recrystallization is-5 ℃～-20 ℃.
Another purpose of the present invention has provided the application in the preparation cancer therapy drug of formula I of the present invention, formula II compound and formula III compound.
It is as follows that the present invention has carried out acute toxicity and the anticancer pharmacodynamics test of above-claimed cpd:
(1) 2, the acute toxicity test (LD of 3,4,6-, four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (III)
50) result.
1) LD as a result after the mouse ig administration
50=2424.0mg.Kg
-1, the 95% credible 1922.1-3057.5mg.Kg that is limited to
-1Dead animal necrotomy result, naked eyes have no the considerable change of the important organs such as the heart, liver, spleen, lung, kidney and gi tract.The surviving animals administration was put to death in 14 days afterwards, and naked eyes have no abnormal changes after the necrotomy.
2) LD as a result after the ip in mice administration
50=384.3mg.Kg
-1, the 95% credible 331.5-445.6mg.Kg that is limited to
-1
(2) 2,3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (I) 2,3,4,6-four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides
(III) antitumor pharmacy test
1) 2,3,4,6-four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (III) is to the restraining effect of animal transplanting tumor growth
Administration group dosage is respectively 0.6g.Kg
-1, 0.3g.Kg
-1, and 0.15g.Kg
-1, positive controls gives endoxan (CTX) 0.025g.Kg
-1, the blank group gives physiological saline.The administration volume is 0.4ml/20g, administration every day one day, and administration is 7 days altogether.Administration finishes 2 days aftertreatment animals.Respectively the restraining effect of Heps.S180 and three kinds of transplanted tumor growths of EC is tested, the results are shown in Table 1,2 and 3.
Administration group dosage is respectively 0.6g.Kg
-1, 0.3g.Kg
-1, and 0.15g.Kg
-1, positive controls gives 10-hydroxycamptothecine with the 0.5mg/kg intravenous injection, and the blank group gives physiological saline.The administration volume is 0.4ml/20g, administration every day one day, and administration is 7 days altogether.Administration finishes 2 days aftertreatment animals.Restraining effect to the growth of Lewis lung cancer kind transplanted tumor is tested, and the results are shown in Table 4.
The administration group has certain growth-inhibiting effect to the MGC803 cell line Nude Mice.When administration group gastric infusion 0.6g/kg, 0.3g/kg, 0.15g/kg dosage, to the tumour inhibiting rate best result Bie Keda 57.3% and 38.9%, 15.4% of MGC803 cell line Nude Mice.When Topotecan is take the 2mg/kg intravenous injection under the similarity condition to the tumour inhibiting rate of MGC803 cell line Nude Mice as 66.5%.The antitumor action of general amine phosphorus is certain dose-effect relationship.The results are shown in Table 5.
The restraining effect of table 1 couple mice-transplanted tumor Heps (
) (n=10)
*P＜0.05
*Compare with the blank group P＜0.01
Table 2 couple mice-transplanted tumor S
180Restraining effect (
) (n=10)
*P＜0.05
*Compare with the blank group P＜0.01
The restraining effect of table 3 couple mice-transplanted tumor EC (
) (n=10)
*P＜0.05
*Compare with the blank group P＜0.01
The restraining effect (X ± sD, n=8) of table 4. pair Lewis lung cancer mouse transplanting tumor growth
*P＜0.05
*Compare with the blank group P＜0.01
The restraining effect (X ± SD, n=8) of table 5. pair MGC803 people's gastric cancer in nude mice xenotransplantation tumor growth
Compare with the blank group,
*P＜0.01
Table 1,2 and 3 show that 2,3,4,6-, four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (III) all has remarkable restraining effect to animal-transplanted tumor Heps.S180e and EC after the ig administration.And at 0.6g.Kg
-1Dosage and positive group (0.025mg.Kg
-1) the same, growth all has remarkable restraining effect to the weight of animals.
Table 4 shows that 2,3,4,6-, four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides also is certain dose-effect relationship to the lung cancer Lewis effect after the ig administration, and at 0.15g.Kg
-1Dosage and positive group (0.5mg.Kg
-1) approach, to the weight of animals is increased remarkable restraining effect is arranged all.
Table 5 shows that 2,3,4,6-, four-0-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (III) has certain growth-inhibiting effect to the MGC803 cell line Nude Mice after the ig administration.During administration 0.6g/kg dosage, to the tumour inhibiting rate best result Bie Keda 57.3% of MGC803 cell line Nude Mice.
(2) 2,3,4,6-, four-O-ethanoyl-α-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (II) is to the restraining effect of growth of tumour cell.
The anti-tumor biological body outer screening test
Screening method: tetrazolium (Methyl-Thiazol-Tetrozlium, MTT reduction method)
Sulphonyl rhodamine B (sulforhodamine B, SRB) albumen beam color method.
Cell strain: HL60 human leukemia
*
Action time: 72 hours
Result evaluation: invalid: 10
-5Mol/L＜85%;
Weak effect: 10
-5Mol/L 〉=85% or 10
-6Mol/L＞50%;
Potent: 10
-6Mol/L 〉=85% or 10
-7Mol/L＞50%.
Inhibiting rate % to growth of tumour cell
Show that according to top test-results 2,3,4,6-, four-O-ethanoyl-α-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (II) is to the HL60 human leukemia
*Have potent.
(3) 2,3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides
(I) to the restraining effect of growth of tumour cell.
The anti-tumor biological body outer screening test
Screening method: tetrazolium (Methyl-Thiazol-Tetrozlium, MTT reduction method)
Sulphonyl rhodamine B (sulforhodamine B, SRB) albumen beam color method.
Cell strain: BEL-7402 people's liver cancer, P338 mouse leukemia *, A-549 human lung adenocarcinoma
Action time: 72 hours
Result evaluation: invalid: 10
-5Mol/L＜85%;
Weak effect: 10
-5Mol/L 〉=85% or 10
-6Mol/L＞50%;
Potent: 10
-6Mol/L 〉=85% or 10
-7Mol/L＞50%.
Inhibiting rate % to growth of tumour cell
Show that according to top test-results 2,3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (I) has potent to BEL-7402 people's liver cancer, P338 mouse leukemia *, A-549 human lung adenocarcinoma.
The present invention is take glucose and 2-chloroethyl amine hydrochloride as main raw material; synthesized 2; 3; 4; 6 four-O-ethanoyl-D-glucopyranosyl-[N; N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (I); this compound comprises α-type (II) and β-type (III) composition; therefore adopt silica gel column chromatography; crystallization and recrystallization have obtained respectively high purity 2,3,4 from synthetic product; 6-four-O-ethanoyl-α-D-glucopyranosyl-[N; N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (II, α-type) and high purity 2,3; 4; 6-four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (III, β-type).Reactions steps of the present invention is few; product yield is high; production cost is low; and this compounds is hydrophobic Ester, is easy to crystallization, chemically with on the zymetology relative stability is arranged all; enter behind the human body by passive diffusion; need not consumed energy can be absorbed by cancer cells, then discharges the different phosphoamide of cell toxicant by esterase, acylations enzyme and glucoside effect in cancer cells, reaches and draws the anticancer growth.
Compound of the present invention can be used as active compound and pharmaceutical excipient is made tablet according to a conventional method, electuary, capsule or injection.
Compound of the present invention can be used for the cancer therapy drugs such as preparation treatment lung cancer, liver cancer, carcinoma of the colon and rectum, cancer of the stomach, oral carcinoma, esophagus cancer, nasopharyngeal carcinoma, He Jiejin lymphatic cancer, prostatic cancer, cancer of pancreas, cervical cancer, non-hodgkin's lymphatic cancer, carcinoma of gallbladder, mammary cancer or leukemia.
Embodiment
Set forth 2 by following instance; 3; the isolation technique of α-type (II) and two kinds of isomer of β-type (III) in the new synthetic technology of 4,6-, four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides and the product.Simultaneously product is carried out ultimate analysis (analyser: Elementar Vario ELIII); Fusing point test (determinator: MetzscH DSC204 or capillary melting point determination instrument); Perkin Elmer214MC) and INFRARED SPECTRUM analysis (analyser: NICOLET impact 410) specific rotation is measured (determinator:
Synthesizing of example one [N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides
Add 1000ml methylene dichloride and 170g 2-chloroethyl amine hydrochloride in the 2000ml three-necked bottle, ice bath is cooled to below-10 ℃, drips the 65ml phosphorus oxychloride.Then after dripping the dichloromethane solution of 500ml 50% triethylamine about 0 ℃ and stirring 2.5 hours, except tetrahydrofuran (THF), stirring reaction is 1.5 hours under the ice bath in 20～25 ℃ of steamings for filtrate, the leaching crystallization, wash successively with cold water, sherwood oil, recrystallizing methanol gets 38.7g, mpl08～110 ℃.The silica gel G thin-layer chromatography (propyl carbinol: acetic acid: water=4: 1: 5, V/V), decide the phosphorus reagent colour development and be blue spot, Rf=0.26～030.
Example 22,3,4,6-, four-O-ethanoyl-α-D-Glucopyranose bromine synthetic
In the 250ml three-necked bottle, add the 120ml aceticanhydride, add 0.4ml perchloric acid and gradation under the ice bath and add altogether 32g of glucose crystallization, in 25～35 ℃ of stirrings 30 minutes, in 18～25 ℃ of dropping 7.8ml phosphorus tribromides, stir after 20 minutes again, add 12ml water, stirred 3 hours under the room temperature, add the 150ml methylene dichloride, use the 200ml water washing, tell organic phase, use 10%NaHCO
3Solution washing is to neutral.Organic addition anhydrous sodium sulfate drying spends the night, and filtering sodium sulfate filters with activated carbon decolorizing, gets 58g with anhydrous diethyl ether crystallization and recrystallization behind the filtrate evaporate to dryness.Mp86～88 ℃; [α]
0 20+ 191.34 ℃ (C3.3, ethanol) [α]
0 20+ 105.86 ℃ (after 24 hours); The silica gel G thin-layer chromatography (propyl carbinol: acetic acid: water=4: 1: 5, V/V), Rf=0.85～0.86.
Example 32, the separation of synthetic and α-type (II) and β-type (III) isomer of 3,4,6-, four-O-ethanoyl-β-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides
In the three-necked bottle of 500ml, add the anhydrous ethylene dichloride of 350ml, 28.5g[N; N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides, 35g 2; 3; 4; 6-four-O-ethanoyl-α-D-Glucopyranose bromine, in 45 ℃ of stirring reactions 24 hours; filter, filtrate is washed successively with 5% sodium chloride solution and cold water.Tell the organic phase activated carbon decolorizing, destainer adds the 70g anhydrous sodium sulfate drying and spends the night, filter, filtrate gets syrup through concentrating under reduced pressure, with silica gel column chromatography (ethylene dichloride: sherwood oil), tell α-type (II) and β-type (III) product, get respectively α-type (II) product 1.4g and β-type (III) product 28.8g through condensing crystal and recrystallization.Their textural property is analyzed as follows:
(1) 2, the textural property analysis of 3,4,6-, four-O-ethanoyl-α-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (II):
Fusing point heat heat is analyzed as unimodal mp91.2 ℃
Specific rotation [α]
D 20-4.820 (C1, ethanol)
Ultimate analysis
IR(KBr)
V
(N-H) 3407.94cm
-1N-H formation vibration frequency; V
(-CH3)2938.63cm
-1-CH
3Stretching vibration frequency absorbs; V
(-CH2-)2888.06cm
-1-CH
2Formation vibration absorbs; V
(C=O)1747.61cm
-1Stretching vibration absorbs; V
(-CH3)1434.23cm
-1-C-H-formation vibration absorbs; V
(-CH3)1382.31cm
-1-CH
3Symmetrical deformation vibration absorbs; V
(C-O-c)1221.36cm
-1Antisymmetric stretching vibration absorbs.
2,2,3,4, the textural property analysis of 6-four-O-ethanoyl-β-D-glucopyranosyl-[N, N`-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides (III):
Silica gel G thin-layer chromatography R=0.84 (propyl carbinol: acetic acid: water=4: 1: 5, V/V), decide the phosphorus reagent colour development.Fusing point: the unimodal mp119.70 of differential thermal analysis ℃,
Specific rotation [α]
D 20-6.31 (C0.4, ethanol)
Ultimate analysis
IR(KBr)
V
(N-H)3393.50cm
-1, 3328.52cm
-1, 3213.00cm
-1N-H symmetry or asymmetrical stretching vibration absorb; V
(CH3 ,-CH2)2945.85cm
-1, 2902.53cm
-1Deng three stretching vibration absorption peaks; V
(C=O)1754.64cm
-1, the C=O stretching vibration absorbs; V
(-CH)1435.25cm
-1,-C-H formation vibration absorbs; V
1369.37Cm
-1The stretching vibration of C-O ester bond absorbs and O=C-CH
3Formation vibration absorbs; V
(O-C=O)1239.51cm
-1, the stretching vibration of O-C=O key absorbs.
Claims (3)
1.2 the preparation method of 3,4,6-, four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides I is characterized in that the method comprises the following steps:
(1) 2, the preparation of 3,4,6-, four-O-ethanoyl-α-D-Glucopyranose bromine V:
Under condition of ice bath, make V with aceticanhydride, glucose and phosphorus tribromide as raw material; The mol ratio of described aceticanhydride, glucose and phosphorus tribromide is 15: 2: 1;
(2) preparation of [N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides VI:
Take phosphorus oxychloride and 2-chloroethyl amine hydrochloride as raw material, under 0 ℃～-20 ℃ conditions, in chloroform, methylene dichloride or ethylene dichloride anhydrous solvent, react, then with in the triethylamine and neutrality make the crystallized product VI; Described phosphorus oxychloride and 2-chloroethyl amine hydrochloride mol ratio are 1.6: 1;
(3) 2,3,4,6-, four-O-ethanoyl-α-D-Glucopyranose bromine prepares I with [N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides:
In anhydrous solvent, 2,3,4,6-, four-O-ethanoyl-α-D-Glucopyranose bromine V carries out condensation reaction and makes I with [N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides VI; Described 2,3,4,6-four-O-ethanoyl-α-D-Glucopyranose bromine V is 1～1.5: 1 with the mol ratio of [N, N '-two (2-chloroethyl)]-di(2-ethylhexyl)phosphate acid amides VI;
(4) the α-type II of separating step (3) condensation product I and β type III:
Condensation product I is through silica gel column chromatography for step (3), methylene dichloride: methyl alcohol=80: 20V/V, 100 milliliters of column volumes, upper column quantity 1%, flow velocity 1～2 ml/min, optically-active according to effluent liquid is different, separates to get α-configuration II and beta configuration III, gets Compound I I and III by crystallization and recrystallization purifying respectively again
Ac is CH in formula I, II, the III
3CO-.
2. according to claim 12,3,4,6-four-O-ethanoyl-D-glucopyranosyl-[N, N '-two (2-chloroethyl)]-preparation method of di(2-ethylhexyl)phosphate acid amides I, it is characterized in that temperature of reaction is 30～70 ℃ in the condensation reaction of step (3) wherein; PH is 6～9; The stirring reaction time is 16～30 hours; Used anhydrous solvent is chloroform, methylene dichloride, 1,2-ethylene dichloride, 1,1-ethylene dichloride, vinyl acetic monomer, tetrahydrofuran (THF), ether, benzene, toluene, sherwood oil or pyridine.
3. according to claim 12; 3; 4; 6-four-O-ethanoyl-D-glucopyranosyl-[N; N '-two (2-chloroethyl)]-preparation method of di(2-ethylhexyl)phosphate acid amides I; the crystallization and the recrystallization solvent that it is characterized in that step (4) formula II wherein or formula III compound are chloroform, methylene dichloride, ethylene dichloride, vinyl acetic monomer, toluene, benzene, tetrahydrofuran (THF), sherwood oil, ether or pyridine or their mixed solvent, and the temperature of crystallization and recrystallization is-5 ℃～-20 ℃.
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| PCT/CN2008/001679 WO2009086720A1 (en) | 2008-01-09 | 2008-09-28 | 2,3,4,6-tetra-o-acetyl-d-glucopyranosyl-[n,n'-di-(2-chloroethyl)]-phosphoric acid diamide, preparation methods and uses thereof |
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