CN102746278A - Chiral pseudonucleoside compound and its preparation method and use - Google Patents

Chiral pseudonucleoside compound and its preparation method and use Download PDF

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CN102746278A
CN102746278A CN2011100979812A CN201110097981A CN102746278A CN 102746278 A CN102746278 A CN 102746278A CN 2011100979812 A CN2011100979812 A CN 2011100979812A CN 201110097981 A CN201110097981 A CN 201110097981A CN 102746278 A CN102746278 A CN 102746278A
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CN102746278B (en
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王梅祥
王德先
陈鹏
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Institute of Chemistry CAS
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Abstract

The invention discloses a chiral pseudonucleoside compound and its preparation method and use. The chiral pseudonucleoside compound is shown in the formula I. The preparation method adopts raw materials prepared by catalytic hydrolysis of an N-heterocyclic diamide compound in the presence of a Rhodococcus erythropolis AJ270 microbial catalytic system. A use amount of Rhodococcus rhodochrous can be adjusted by a use amount of a substrate. A reaction solvent is a common buffer solution having a pH value of 6.0 to 8.0. A reaction temperature is in a range of 20 to 37 DEG C. Reaction time is in a range of 0.1 to 120 hours. The Rhodococcus erythropolis AJ270 microbial catalytic system is suitable for fermentation culture and can be preserved conveniently. The preparation method of a chiral N-heterocyclic diamide, a monoamido carboxylic acid and a dicarboxylic acid is based on biotransformation, has the characteristics of simple processes, high reaction efficiency, mild reaction conditions, high enantioselectivity, easy separation of products and high product purity, and can be used for synthesis of the chiral pseudonucleoside compound which is a novel pseudonucleoside shown in the formula I.

Description

Pseudo-nucleoside compound of chirality and preparation method thereof and application
Technical field
The present invention relates to pseudo-nucleoside compound of chirality and preparation method thereof and application.
Background technology
Pseudo-nucleoside compound; Also claim the non-natural nucleoside compounds; Be one type of compound similar with natural nucleus glycoside on chemical structure, thereby can disturb or directly be used for the metabolic process of nucleic acid, the biosynthesizing of blocking protein, nucleic acid in vivo; Therefore, these compounds occupy an important position in antiviral and antitumor drug.
It is raw material that the compound of present pseudo-ucleosides mainly adopts the natural sugar compounds, synthesizes pseudo-nucleoside compound through the formation of glycosidic link.Wherein the selective control of glycosidic link is relatively more difficult.The present invention makes up cis or the trans false sugar ring of chirality skeleton earlier through the method for biocatalysis, has avoided the selective control of glycosidic link.
Biocatalysis is that up to now tool is efficient, highly selective and eco-friendly process, utilize the method for biocatalysis more synthetic have the chemical of high added value, particularly the chirality chemical has important application prospects and meaning.Nitrile is one type of important organic synthesis intermediate; The chemical conversion requirement condition harshness and the selectivity of nitrile are very poor; And the bioconversion reaction of nitrile has advantages such as mild condition, highly selective; Applicable industryization prepares corresponding carboxylic acid and amide derivatives at present, and foremost is to be taken the lead in realizing the industriallization of microbial method synthesis of acrylamide in 1985 by Japanese Nitto company (renaming Mitsubishi Rayon company as) at present.The YO of acrylic amide in 1998 surpasses 40,000 tons, becomes one of industrial biological path for transformation of present maximum-norm in the world.Rhodococcus Rhodococcus rhodochrous J1 also by Switzerland Lonza AG company in order to suitability for industrialized production vitamin B group vitamin PP and nicotinic acid, wherein the YO of vitamin PP is above 3000 tons.But form the reacting phase ratio with hydrolysis of enzyme catalysis ester and ester bond, the biocatalytic reaction of nitrile and acid amides is also less relatively.The bioconversion reaction of nitrile and acid amides can not only obtain carboxylic acid, and through kinetic resolution with go method of symmetrization can also obtain unavailable organic compounds containing nitrogen acid amides of ester hydrolysis reaction or nitrile.
Summary of the invention
The purpose of this invention is to provide pseudo-nucleoside compound of a kind of chirality and preparation method thereof and application.
The pseudo-nucleoside compound of chirality provided by the invention is compound shown in the formula I;
Figure BDA0000056120000000011
(formula I)
Among the said formula I, n is the integer of 1-3, and m is 0 or 1;
* represent chirality, be R or S configuration;
X is-OH,
Figure BDA0000056120000000012
Wherein, R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
Y is-OH,
Figure BDA0000056120000000013
Wherein, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN.
M is 0 in shown in the above-mentioned formula I of preparation provided by the invention, Y is the method for
Figure BDA0000056120000000021
X for the pseudo-nucleoside compound of-OH,
Figure BDA0000056120000000022
; Be method a or method b; Wherein, Method a is an initial reactant with compound shown in the formula II, comprises the steps:
Figure BDA0000056120000000024
1) with said compound of formula II ' and SOCl 2In N, to react in the dinethylformamide, reaction finishes and obtains compound shown in the formula IIa1;
(formula II ')
Among the said formula II ', n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN. R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
Figure BDA0000056120000000026
(formula IIa1)
2) with compound and NaN shown in the step 1) gained formula IIa1 3And ZnBr 2In the mixed solution of being made up of Virahol and water, carry out back flow reaction, reaction finishes and obtains compound shown in the formula IIb1;
Figure BDA0000056120000000031
(formula IIb1)
3) with step 2) compound and LiAlH shown in the gained formula IIb1 4In THF, carry out back flow reaction, reaction finishes and obtains compound shown in the formula IIc1;
(formula IIc1)
4) compound shown in the formula IIc1 of step 3) gained is reacted with diacetyl oxide, pyridine and 4-dimethylamino-pyridine in methylene dichloride; Reaction finish obtain that m among the said formula I is 0, Y is for the pseudo-nucleoside compound of
Figure BDA0000056120000000033
X for-OH,
Figure BDA0000056120000000034
Figure BDA0000056120000000035
, also is compound shown in the formula IId1;
Figure BDA0000056120000000036
(formula IId1)
Among said formula IIa1, formula IIb1, formula IIc1 and the formula IId1, n is the integer of 1-3; * represent chirality, be R or S
Figure BDA0000056120000000037
In the said step 1) of this method, the said compound of said formula II ', SOCl 2With N, the amount ratio of dinethylformamide is 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferred 10mmol: 1mL: 4mL; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
Said step 2) in, compound, NaN shown in the said formula IIa1 3, ZnBr 2With the amount ratio of the said mixed solution of forming by Virahol and water be 0.1-10.0mmol: 0.2-20mmol: 0.1-10mmol: 1-10mL, preferred 1mmol: 2mmol: 0.5mmol: 4.5mL; In the said mixed solution of being made up of Virahol and water, the volume ratio of Virahol and water is 1-10: 0.1-100, preferred 1: 2; In the said back flow reaction step, the time is 1-24 hour, preferred 16 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 3), compound, LiAlH shown in the said formula IIb1 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 1mmol: 10mmol: 10mL; In the said back flow reaction step, the time is 6-48 hour, preferred 12 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 4); The amount ratio of compound, diacetyl oxide, pyridine, 4-dimethylaminopyridine, methylene dichloride shown in the said formula IIc1 is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
Method b is with formula III ' shown in compound be initial reactant, comprise the steps:
Figure BDA0000056120000000041
Said method b comprises the steps:
1) with formula III ' said compound and SOCl 2In DMF, react, reaction finishes and obtains compound shown in the formula III a1;
(formula III ')
Said formula III ' in, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3, R 3' be-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
Figure BDA0000056120000000043
(formula III a1)
2) with compound and NaN shown in the step 1) gained formula III a1 3And ZnBr 2In the mixed solution of being made up of Virahol and water, carry out back flow reaction, reaction finishes and obtains compound shown in the formula III b1;
Figure BDA0000056120000000051
(formula III b1)
3) with step 2) compound and LiAlH shown in the gained formula III b1 4In THF, carry out back flow reaction, reaction finishes and obtains compound shown in the formula III c1;
Figure BDA0000056120000000052
(formula III c1)
4) compound shown in the formula III c1 of step 3) gained is reacted with acetic anhydride, pyridine and 4- dimethylamino-pyridine in carrene; Reaction finish obtain that m among the said formula I is 0, Y is the pseudo- nucleoside compound of
Figure BDA0000056120000000053
X for-OH,
Figure BDA0000056120000000054
Figure BDA0000056120000000055
; It also is compound shown in the formula III d1
Figure BDA0000056120000000056
(formula III d1)
Among said formula III a1, formula III b1, formula III c1 and the formula III d1, n is the integer of 1-3; * represent chirality, be R or
Figure BDA0000056120000000057
In the step 1) of aforesaid method, said formula III ' said compound, SOCl 2With the amount ratio of DMF be 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferred 10mmol: 1mL: 4mL; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
Said step 2) in, compound, NaN shown in the said formula III a1 3, ZnBr 2With the amount ratio of the said mixed solution of forming by Virahol and water be 0.1-10.0mmol: 0.2-20mmol: 0.1-10mmol: 1-10mL, preferred 1mmol: 2mmol: 0.5mmol: 4.5mL; In the said mixed solution of being made up of Virahol and water, the volume ratio of Virahol and water is 1-10: 0.1-100, preferred 1: 2; In the said back flow reaction step, the time is 1-24 hour, preferred 16 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 3), compound, LiAlH shown in the said formula III b1 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 1mmol: 10mmol: 10mL; In the said back flow reaction step, the time is 6-48 hour, preferred 12 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 4); The amount ratio of compound, diacetyl oxide, pyridine, 4-dimethylaminopyridine, methylene dichloride is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL shown in the said formula III c1, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours.According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
M is 1 in shown in the above-mentioned formula I of preparation provided by the invention, Y is for-OH or
Figure BDA0000056120000000061
X method for the pseudo-nucleoside compound of-OH,
Figure BDA0000056120000000062
, comprises the steps:
1) with said compound of formula II ' and LiAlH 4In THF, carry out back flow reaction, reaction finishes and obtains compound shown in the formula IIa2;
Figure BDA0000056120000000064
(formula II ')
Among said formula II ' and the formula IIa2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
Figure BDA0000056120000000071
(formula IIa2)
2) with compound and R shown in the said step 1) gained formula IIa2 4PhCH 2CH 2CHO and NaBCNH 3In methylene dichloride or methyl alcohol, react, reaction finishes and obtains compound shown in the formula IIb2; Said R 4PhCH 2CH 2Among the CHO, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
Figure BDA0000056120000000072
(formula IIb2)
Among the said formula IIb2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN. R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a; Ph is a phenyl;
3) with said step 2) aqueous solution of compound shown in the gained formula IIb2 and hydrochloric acid and formaldehyde carries out back flow reaction; Reaction finishes and obtains compound shown in the formula IIc2, also is that m is 1 among the said formula I, Y is-OH, the compound of X for
Figure BDA0000056120000000073
;
(formula IIc2)
Among the said formula IIc2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN. R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
4) compound shown in the formula IIc2 of step 3) gained is reacted with acetic anhydride, pyridine and 4- dimethylamino-pyridine in carrene; Reaction finish obtain that m among the said formula I is 1, Y is the pseudo- nucleoside compound of
Figure BDA0000056120000000075
X for-OH,
Figure BDA0000056120000000076
Figure BDA0000056120000000081
; It also is compound shown in the formula IId2
(formula IId2)
Among the said formula IId2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN. R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a; X is-OH,
Figure BDA0000056120000000083
R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5.
In the step 1) of aforesaid method, the said compound of said formula II ', LiAlH 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 1mmol: 10mmol: 10mL; In the said back flow reaction step, the time is 6-48 hour, preferred 24 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
Said step 2) in, compound, R shown in the said formula IIa2 4PhCH 2CH 2CHO, NaBCNH 3With the amount ratio of methylene dichloride or methyl alcohol be 0.1-10mmol: 0.1-10mmol: 1-100mmol: 1-100mL, preferred 1mmol: 0.9mmol: 2mmol: 6mL; In the said reactions step, temperature is 0-50 ℃, and preferred 25 ℃, the time is 12-48 hour, preferred 24 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 3), the amount ratio of the aqueous solution of compound, formaldehyde, hydrochloric acid and chloroform shown in the said formula IIb2 is 0.1-10mmol: 0.1-20mL: 1-40mL: 0-40mL, preferred 1mmol: 2.4mL: 4.8mL: 4.8mL; In the said back flow reaction step, the time is 1-48 hour, preferred 24 hours; The mass percentage concentration of said hydrochloric acid is 10-35%, preferred 35%; The mass percentage concentration of the aqueous solution of said formaldehyde is 10-35%, preferred 35%; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 4); Compound shown in the formula IIc2, diacetyl oxide, pyridine, 4-dimethylaminopyridine, methylene dichloride amount ratio are 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours.According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
M=1 among the above-mentioned formula I of preparation provided by the invention; X is-OH,
Figure BDA0000056120000000084
Y be the method for the pseudo-nucleoside compound of-OH or , comprise the steps:
1) the said compound of formula II ' and dimethoxy Cynuric Chloride, nitrogen methylmorpholine and tetrahydroisoquinoline are reacted in methylene dichloride, reaction finishes and obtains compound shown in the formula IIa3;
Figure BDA0000056120000000092
(formula II ')
Among said formula II ' and the formula IIa3, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
(formula IIa3)
2) with compound and SOCl shown in the step 1) gained formula IIa3 2In DMF, react, reaction finishes and obtains compound shown in the formula IIb3;
Figure BDA0000056120000000101
(formula IIb3)
3) with step 2) compound shown in the gained formula IIb3 and the mixed solution of forming by methyl alcohol and methylene dichloride, feed dry HCl gas, react, reacting finishes obtains compound shown in the formula IIc3;
(formula IIc3)
4) with compound and LiAlH shown in the step 3) gained formula IIc3 4In THF, carry out back flow reaction, reaction finishes and obtains compound shown in the formula IId3, also is that m=1 among the said formula I, X do
Figure BDA0000056120000000103
Y is-compound of OH;
(formula IId3)
5) compound shown in the formula IId3 of step 3) gained is reacted with acetic anhydride, pyridine and 4- dimethylamino-pyridine in carrene; Reaction finishes and obtains m=1 among the said formula I; X is-OH, the pseudo- nucleoside compound of Y for
Figure BDA0000056120000000106
; It also is compound shown in the formula IIe3
Figure BDA0000056120000000111
(formula IIe3)
Among the said formula IIb3-formula IIe3, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; X is-OH,
Figure BDA0000056120000000112
R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a.
In the step 1) of aforesaid method; The amount ratio of the said compound of said formula II ', CDMT (dimethoxy Cynuric Chloride), NMM (nitrogen methylmorpholine), tetrahydroisoquinoline and methylene dichloride is 0.1-10mmol: 0.1-20mmol: 0.1-20mmol: 0.1-10mmol: 1-40mL, preferred 5mmol: 1.2mmol: 3mmol: 1.1mmol: 10mL; In the said reactions step, temperature is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
Said step 2) in, compound, SOCl shown in the said formula IIa3 2With the amount ratio of DMF be 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferred 4mmol: 1mL: 4mL; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 3), compound shown in the said formula IIb3, the mixed solution amount ratio of being made up of methyl alcohol and methylene dichloride are 0.1-1mmol: 1-15mL, preferred 0.5mmol: 15mL; The feeding volume of said dry HCl gas is 22.4mL-22.4L, preferred 1L; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour; In the said mixed solution of being made up of methyl alcohol and methylene dichloride, the volume ratio of methyl alcohol and methylene dichloride is 2: 1; In the said reactions step, temperature is-20-0 ℃, and preferred-20 ℃, the time is 0.5-12 hour, preferred 1 hour; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 4), compound, LiAlH shown in the said formula IIc3 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 0.1mmol: 10mmol: 4mL; In the said back flow reaction step, the time is 6-48 hour, preferred 24 hours; According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
In the said step 5); Compound shown in the formula IId3, diacetyl oxide, pyridine, 4-dimethylaminopyridine, methylene dichloride amount ratio are 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours.According to above-mentioned optimal conditions reaction scale scope is 0.5mmol-100mmol;
The present invention also provides compound shown in the starting material compound formula II for preparing compound shown in the above-mentioned formula I and the formula III, and compound is following shown in its Chinese style II,
Figure BDA0000056120000000121
(formula II)
Among the said formula II, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN. R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
Compound shown in the said formula II is a compound shown in the formula II ',
Figure BDA0000056120000000122
(formula II ')
Among the said formula II ', n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
The method of compound shown in the above-mentioned formula II of preparation provided by the invention is through going symmetrization catalytic hydrolysis reaction and esterification to get, and its reaction equation is following:
Figure BDA0000056120000000123
(formula IV)
This preparation method is following method a or method b; Wherein, After said method a comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula IV; In solvent, carry out esterification with alkali and benzyl bromine or to the methoxybenzyl bromine again, reaction finishes and obtains compound shown in the said formula II
Figure BDA0000056120000000124
(formula IV)
Among the said formula IV, R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3;
After said method b comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula IV; Diethyl ether solution with diazomethane carries out esterification again; Reaction finishes and obtains compound shown in the said formula II
Figure BDA0000056120000000131
(formula IV)
Among the said formula IV, R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3.
Among said method a and the method b, said rhodococcus catalystsystem is that the buffered soln of 6.0-8.0 is formed by rhodococcus and pH value, is specially said rhodococcus is connected to said pH value got in 30 minutes for 30 ℃ of activation in the buffered soln of 6.0-8.0; Said buffered soln is Na 2HPO 4-citric acid solution, K 2HPO 4-KH 2PO 4Buffered soln, Tris buffered soln, Hanks ' buffered soln or PBS buffered soln; The amount ratio of compound is 2g: 1mmol-2g: 1mol shown in said rhodococcus and the formula IV; In the said rhodococcus catalystsystem, the amount ratio of rhodococcus and said buffered soln is 2g: 50mL-2g: 1L; In the said catalytic hydrolysis reaction step, temperature is 20-37 ℃, and preferred 30 ℃, the time is 0.1-120 hour, and the preferred different time of different substrates and consumption makes the reaction product enantioselectivity at more than 95% hour;
Among the said method a, said alkali is salt of wormwood, yellow soda ash, Pottasium Hydroxide, sodium hydroxide or cesium carbonate, preferred salt of wormwood; Said solvent is selected from acetone, N, at least a in dinethylformamide, DMSO 99.8MIN. and the THF, preferred N, dinethylformamide; Said alkali, said solvent, benzyl bromine or be 1.38g: 2mL: 1mL: 1mmol to the amount ratio of compound shown in methoxybenzyl bromine and the said formula IV; In the said step of esterification, temperature is 10-30 ℃, and preferred 25 ℃, the time is 1-48 hour, preferred 24 hours;
Among the said method b, the concentration of the diethyl ether solution of said diazomethane is 0.1-lmol/L, preferred 0.5mol/L; In the said step of esterification, temperature is-20-10 ℃, and preferred 0 ℃, the time is 2-24 hour, preferred 12 hours.
The present invention also provides compound shown in the formula III,
Figure BDA0000056120000000132
(formula III)
In the said formula III, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN. R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3, R 3' be-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
The said compound of said formula III is a formula III ' shown in compound,
Figure BDA0000056120000000141
(formula III ')
Said formula III ' in, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3, R 3' be-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
The method of compound shown in the above-mentioned formula III of preparation provided by the invention is to get through kinetic resolution and esterification, and its reaction equation is following:
Figure BDA0000056120000000142
This preparation method is following method a or method b; Wherein, After said method a comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula V; In solvent, carry out esterification with alkali and methyl iodide, benzyl bromine or to the methoxybenzyl bromine again, reaction finishes and obtains compound shown in the said formula III
Figure BDA0000056120000000143
(formula V)
Among the said formula V, R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3;
After said method b comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula V; Diethyl ether solution with diazomethane carries out esterification again; Reaction finishes and obtains compound shown in the said formula III
Figure BDA0000056120000000151
(formula V)
Among the said formula V, R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3, Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3.
Among said method a and the method b, said rhodococcus catalystsystem is that the buffered soln of 6.0-8.0 is formed by rhodococcus and pH value, is specially said rhodococcus is connected to said pH value got in 30 minutes for 30 ℃ of activation in the buffered soln of 6.0-8.0; Said buffered soln is Na 2HPO 4-citric acid solution, K 2HPO 4-KH 2PO 4Buffered soln, Tris buffered soln, Hanks ' buffered soln or PBS buffered soln; The amount ratio of compound is 2g: 1mmol-2g: 1mol shown in said rhodococcus and the formula IV; In the said rhodococcus catalystsystem, the amount ratio of rhodococcus and said buffered soln is 2g: 50mL-2g: 1L; In the said catalytic hydrolysis reaction step, temperature is 20-37 ℃, and preferred 30 ℃, the time is 0.1-120 hour, and the preferred different time of different substrates and consumption makes the reaction product enantioselectivity more than 95%;
Among the said method a, said alkali is salt of wormwood, yellow soda ash, Pottasium Hydroxide, sodium hydroxide or cesium carbonate, preferred salt of wormwood; Said solvent is selected from acetone, N, at least a in dinethylformamide, DMSO 99.8MIN. and the THF, preferred N, dinethylformamide; Said alkali, said solvent, benzyl bromine or be 1.38g: 2mL: 1mL: 1mmol to the amount ratio of compound shown in methoxybenzyl bromine and the said formula IV; In the said step of esterification, temperature is 10-30 ℃, and preferred 25 ℃, the time is 1-48 hour, preferred 24 hours;
Among the said method b, the concentration of the diethyl ether solution of said diazomethane is 0.1-1mol/L, preferred 0.5mol/L; In the said step of esterification, temperature is-20-10 ℃, and preferred-15 ℃, the time is 2-24 hour, preferred 12 hours.
Biocatalysis substrate described in above-mentioned two microorganism catalysis-esterification process (substrate) preparation method can be divided into following three types according to literature reference.
The first kind: prepare cis shown in the above-mentioned formula IV and formula V trans-N-replaces 2,5-two substituted-tetrahydro pyrroles diamide compound, wherein n=1.This compounds is by 2, the preparation of 5-two substituted-tetrahydro pyrroles dinitrile compound chemical hydrolysis.Nitrile compounds is to prepare according to the method that following document provides: Takahashi, K.; Saitoh, H.; Ogura, K.; Iida, H.Heterocycles1986,24,10,2905.
Second type: it is trans-2 to prepare cis shown in the above-mentioned formula IV and formula V, and 6-two replaces six hydrogen piperidines diamide compound, wherein n=2.Prepare according to following literature method: Takahashi K.; Mikajiri T.; Kurita H.; Ogura K.; Iida H., J.Org.Chem.1985,50,4372-4375.
The 3rd type: it is trans-2 to prepare cis shown in the above-mentioned formula IV and formula V, and 7-two replaces nitrogen seven-membered ring diamide compound, wherein n=3.According to literature method: Ogura K.; Shimamura Y.; Fujita M., J.Org.Chem.1991,56,2920-2922.
Employed rhodococcus Rhodococcus erythropolis AJ270 sample is to use at first in the Anderson biomone sampling thief to separate from the acetonitrile nutrient agar that contains 25mM and obtains in above-mentioned two microorganism catalysis-esterification process, and sample source then is collected near the exsiccant soil Britain's Tyne side depleted industrial working at first.Mycolic acids and the research of diaminopimelic acid chemotaxonomy to its cell walls of AJ270 bacterium have confirmed that it belongs to the Rhodococcus bacterial classification.Up to 2005,, confirmed that RhodococcusAJ 270 belongs to Rhodococcus erythropolis fungus strain through research to its 16SrRNA gene order.Concrete with reference to following two pieces of documents:
a.Blakey?A.J.；Colby?J.；Williams?E.；O’Reilly?C.，FEMS?Microbiol.Lett.1995，129，57-61.
b.O’Mahony?R.；Doran?J.；Coffey?L.；Cahill?O.J.；Black?G.W.；O’Reilly?C.；Antonie?vanLeeuwenhoek?2005，87，221-232.
Rhodococcus AJ 270 is a kind of mikrobes that are derived from soil, and is proved to be a kind of highly active intact cell catalysis agent that contains Nitrile hydratase/amide hydrolysis enzyme system.Existing research shows, compares with other bacterial strains, and rhodococcus Rhodococcuserythropolis AJ270 has good substrate broad spectrum, efficiently the hydrolysis of catalysis fatty nitrile, fragrant nitrile and fragrant heterocycle nitrile compounds.(Wang?M.-X.Enantioselective?biotransformations?of?nitrile?in?organic?synthesis.Top.Catal.2005，35，117-130)
In addition, the antitumor drug that is activeconstituents with pseudo-nucleoside compound provided by the present invention belongs to protection scope of the present invention.In actually operating,, can in above-mentioned pseudo-nucleoside compound, add various assistant agents as required for reaching better drug effect.In addition, the application of pseudo-nucleoside compound provided by the present invention in the preparation antitumor drug also belongs to protection scope of the present invention.Said tumour is liver cancer or lung cancer.Above-mentioned antitumor drug can be the medicine of anti-Bel-7402 liver cancer cell or A-549 lung carcinoma cell.
The raw material of the pseudo-nucleoside compound of preparation provided by the invention is to obtain with rhodococcus Rhodococcus erythropolis AJ270 microorganism system catalytic hydrolysis azepine ring-type diamide compound.Used rhodococcus thalline consumption can be regulated according to the consumption of substrate.Reaction solvent is the buffered soln commonly used of pH=6.0-8.0, and temperature is 20-37 ℃, and the reaction times is 0.1-120 hour.But this rhodococcus microorganism catalysis system has fermentation culture and preserves characteristics easily.Use this bio-transformation to prepare the method for chirality azepine ring-type diamide, monoamide carboxylic acid, dicarboxylicacid, have easy and simple to handlely, reaction is efficient; Reaction conditions is gentle, and enantioselectivity is high, and product is easily separated; The characteristics that product purity is high, and synthesizing as novel pseudo-nucleosides.
Embodiment
Below in conjunction with specific embodiment the present invention is done further elaboration, but the present invention is not limited to following examples.Said method is ordinary method if no special instructions.Said reactant all can get from open commercial sources if no special instructions.The Chinese of Cbz is carbobenzoxy-(Cbz) among the present invention.
Embodiment 1: pseudo-nucleoside compound 1 ' synthetic
Figure BDA0000056120000000161
1) with compounds X shown in the embodiment 15 preparation gained formula II ' ' (Cbz, Chinese carbobenzoxy-(Cbz)) 10mmol is dissolved among 4 milliliters of dry DMF, adds 1 milliliter of thionyl chloride SOCl 2, 0 ℃ was stirred 1 hour down, disappears according to TLC monitoring reaction raw material.Reaction solution is through ethyl acetate extraction and use the ETHYLE ACETATE silica gel column chromatography, obtains the cyanic acid of compound list shown in formula IIa1 monocarboxylate (also being compound shown in the 1-1 in the reaction formula), and oily liquids, productive rate are 90%.
2) take out 2mmol step 1) gained list cyanic acid monocarboxylate and put in 25 milliliters of eggplant-shape bottles, add 4mmol sodium azide NaN 3, 1mmol zinc bromide ZnBr 2And by 6 ml waters and 3 milliliters of mixed solutions that Virahol is formed, back flow reaction 16 hours.After the cooling, regulate pH less than 7 with 1N HCl, ethyl acetate extraction.Use silica gel column chromatography then, leacheate is an ETHYLE ACETATE: methyl alcohol=20: 1 (volume ratio, 1% acetic acid is as additive); Receiving colorless oil, be compound shown in the formula IIb1, (also is compound shown in the 1-2 in the reaction formula; This compound is the precursor of compound 1), productive rate is 58%.
3) get 1mmol step 2) compound shown in the gained 1-2, add 10mmol Li-Al hydrogen and 10mL THF and refluxed 12 hours, obtain compound shown in the formula IIc1 (also being compound 1 in the reaction formula) through the Zeo-karb purifying, be white solid, productive rate 83%.
Compound 1:solid, 185 ℃ of (D of mp 2O/300MHz) δ 3.86-3.75 (m, 2H), 3.64-3.58 (m, 1H), 2.80-2.71 (m, 1H), 2.32 (m, 3H), 2.27-2.14 (m, 3H), 1.95-1.77 (m, 1H); (75MHz/D 2O) δ 163.0,67.6,63.6,62.5,39.5,28.6,26.8. (KBr) 3432,3259,1634,1023cm -1MS (ESI) m/z184.1 (M ++ 1,15%) 206.1 (M, ++ 23,100%) .Anal.Calcd.for: [M+H] C 7H 14N 5O, 184.1198, Found: [M+H] 184.1193.
By on can know that this compound structure is correct, be compound in the reaction formula 1.
4) step 3) gained compound 1 is got 1mmol, be dissolved in the 10mL methylene dichloride, add the 10mmol diacetyl oxide, the 1mmol pyridine; 0.1mmol 4-dimethylaminopyridine stirred 12 hours at 25 ℃, obtained pseudo-nucleoside compound 1 ' shown in the formula I provided by the invention, wherein; N is 1, and m is 0, and Y does
Figure BDA0000056120000000171
R 2For-CH 3, X is-OCOCH 3
Compound 1 ': solid, 105 ℃ of (D of mp 2O/300MHz) δ 3.91-3.70 (m, 2H), 3.60-3.45 (m, 1H), 2.88-2.81 (m, 1H), 2.35 (m, 3H), 2.22-2.10 (m, 6H), 2.01-1.75 (m, 1H); (75MHz/D 2O) δ 162.5,65.6,62.6,61.5,39.2,28.4,27.2,20.7. (KBr) 3432,3259,1634,1023cm -1Anal.Calcd.for: [M+H] C 9H 15N 5O 2, 226.1304, Found: [M+H] 226.1300.
By on can know that this compound structure is correct, be compound shown in the formula I, also be compound 1 ' in the reaction formula.
Embodiment 2: pseudo-nucleoside compound 2 ' synthetic
1) with embodiment 8 preparation gained formula IIIs ' shown in compounds X ' ' (Cbz, Chinese carbobenzoxy-(Cbz)) 10mmol is dissolved among 4 milliliters of dry DMF, adds 1 milliliter of thionyl chloride SOCl 2, 0 ℃ was stirred 1 hour down, disappears according to TLC monitoring reaction raw material.Reaction solution is through ethyl acetate extraction and use the ETHYLE ACETATE silica gel column chromatography, obtains the cyanic acid of compound list shown in formula III a1 monocarboxylate (also being compound 2-1 in the reaction formula), and oily liquids, productive rate are 90%.
2) take out 2mmol step 1) gained list cyanic acid monocarboxylate and put in 25 milliliters of eggplant-shape bottles, add 4mmol sodium azide NaN 3, 1mmol zinc bromide ZnBr 2And the mixed-liquor return reaction of forming by 6 ml waters and 3 milliliters of Virahols 16 hours, after the cooling, with 1N HCl adjusting pH less than 7, ethyl acetate extraction.Use silica gel column chromatography then; Leacheate is an ETHYLE ACETATE: methyl alcohol=20: 1 (1% acetic acid is as additive), receive the white solid thing, for compound shown in the formula III b1 (also is a compound shown in the 2-2 in the reaction formula; This compound is the precursor of compound 2), productive rate is 90%.
3) get 1mmol step 2) compound shown in the gained 2-2, add the 10mmol Li-Al hydrogen, the 10mL THF refluxed 12 hours, obtained compound shown in the formula III c1 (also being compound 2 in the reaction formula) through the Zeo-karb purifying, was white solid, productive rate 83%.
Compound 2:solid, 194 ℃ of (D of mp 2O/300MHz) δ 4.91-4.88 (m, 1H), 3.76-3.75 (m, 2H), 3.47-3.44 (m, 1H), 2.43-2.26 (m, 3H), 2.30 (s, 3H), 1.97-1.86 (m, 1H); (75MHz/D 2O) δ 158,67.0,60.7,59.3,35.7,27.4,25.5. (KBr) 3432,3259,1634,1023cm -1MS (ESI) m/z184.1 (M ++ 1,15%) 206.1 (M, ++ 23,100%) .Anal.Calcd.for: [M+H] C 7H 14N 5O, 184.1198, Found: [M+H] 184.1193.
By on can know that this compound structure is correct, be compound in the reaction formula 2.
4) get step 3) gained compound 21mmol, be dissolved in the 10mL methylene dichloride, add the 10mmol diacetyl oxide, the 1mmol pyridine; 0.1mmol 4-dimethylaminopyridine stirred 12 hours at 25 ℃, obtained pseudo-nucleoside compound 2 ' shown in the formula I provided by the invention, wherein; N is 1, and m is 0, and Y does
Figure BDA0000056120000000182
R 2For-CH 3, X is-OCOCH 3
Compound 2 ': solid, 154 ℃ of (D of mp 2O/300MHz) δ 4.77-4.50 (m, 1H), 3.86-3.75 (m, 2H), 3.52-3.34 (m, 1H), 2.33-2.16 (m, 6H), 2.30 (s, 3H), 1.97-1.86 (m, 1H); (75MHz/D 2O) δ 155,67.5,60.2,58.3,33.7,27.3,24.5,20.7. (KBr) 3432,3259,1634,1023cm -1Anal.Calcd.for: [M+H] C 9H 15N 5O 2, 226.1304, Found: [M+H] 226.1307.
By on can know that this compound structure is correct, be compound shown in the formula I (also being compound 2 in the reaction formula).
Embodiment 3: pseudo-nucleoside compound 3 ' shown in the formula I synthetic
1) with compounds X shown in the 5mmol embodiment 15 preparation gained formula II ' ' be dissolved in 50 milliliters of THFs; Add Li-Al hydrogen 1.9 grams (50mmol), reflux after 24 hours, add 2N aqueous sodium hydroxide solution cancellation reaction; Through the zeyssatite suction filtration; The Zeo-karb purifying obtains primary amine shown in the formula IIa2 (being specially compound shown in the 3-1 in the above-mentioned reaction equation), productive rate 79%.
2) primary amine 0.625mmol shown in the step 1) gained formula IIa2 and 0.6mmol phenylacetic aldehyde are blended in 4 milliliters of methylene dichloride; Add the 1.2mmol sodium cyanoborohydride; 25 ℃ of reactions are after 24 hours; With 1N HCl product is extracted from organic phase, regulate pH then, product is extracted from water with ETHYLE ACETATE greater than 7.Further analyse and obtain compound shown in the formula IIb2 (being specially compound shown in the 3-2 in the above-mentioned reaction equation), productive rate 54% with cation exchange resin layer.
3) get step 2) compound 0.5mmol shown in the gained formula IIb2, add 1.2 milliliters of mass percentage concentration and be 35% formalin, 2.4 milliliters of mass percentage concentration are 35% concentrated hydrochloric acid; 2.4 the milliliter chloroform, after the reaction in 24 hours that refluxes finished, regulation system pH value was 9; Dichloromethane extraction; Anhydrous sodium sulfate drying, the conventional column chromatography of ODS, moving phase is methyl alcohol: water=1: 1; Chromatography finishes and obtains compound shown in the formula IIc2 (being specially compound shown in 3 in the above-mentioned reaction equation), productive rate 78%.
Compound 3:oil, 1HNMR (D 2O/300MHz) δ 7.11-6.99 (m, 4H), 3.63-3.57 (m, 3H), 3.63-3.33 (m, 1H), 2.84-2.72 (m, 3H), 2.69-2.37 (m, 2H), 2.28 (s, 3H), 2.10-2.02 (m, 1H), 1.95-1.79 (m, 1H), 1.51-1.38 (m, 2H); 13CNMR (75MHz/CDCl 3) δ 134.9,134.4,128.7,126.5,126.1,125.6,67.7,64.7,63.5,61.1,56.99,39.99,29.8,29.1,26.1; IR (KBr) 3411,2934,1641,1450,1050cm -1MS (ESI) m/z 261.4 (M ++ 1,100%) Anal.Calcd.for [M+H] C, 16H 25N 2O, 261.1967, Found: [M+H] 261.1961.
By on can know that this compound structure is correct, be compound in the reaction formula 3.
4) compound 1mmol shown in the step 3) gained formula IIc2 is dissolved in the 10mL methylene dichloride; Add the 10mmol diacetyl oxide, 1mmol pyridine, 0.1mmol 4-dimethylaminopyridine; 25 ℃ of stirring reactions 12 hours, reaction finished and obtains pseudo-nucleoside compound 3 ' shown in the formula I provided by the invention.
In the pseudo-nucleoside compound 3 ', n is 1, R shown in this formula I 2For-CH 3, m is 1, Y does
Figure BDA0000056120000000201
Wherein, R 4Be H, X is-OCOCH 3
Compound 3 ': oil, 1HNMR (D 2O/300MHz) δ 7.21-6.95 (m, 4H), 3.65-3.52 (m, 3H), 3.67-3.31 (m, 1H), 2.85-2.70 (m, 3H), 2.67-2.33 (m, 2H), 2.28 (s, 3H), 2.10-2.02 (m, 4H), 1.95-1.79 (m, 1H), 1.51-1.38 (m, 2H); 13CNMR (75MHz/CDCl 3) δ 134.5,134.7,128.3,126.4,126.1,122.6,67.7,62.7,61.5,60.1,56.4,39.5,29.7,28.1,22.1,21.9; IR (KBr) 3411,2934,1641,1450,1050cm -1MS (ESI) m/z 261.4 (M ++ 1,100%) Anal.Calcd.for [M+H] C, 18H 26N 2O 2, 303.2073, Found: [M+H] 303.2070.
By on can know that this compound structure is correct, be compound shown in the formula I.
Embodiment 4: pseudo-nucleoside compound 4 ' shown in the formula I synthetic
1) with compounds X shown in the 5mmol embodiment 15 preparation gained formula II ' ' be dissolved in 10 milliliters of methylene dichloride; Add 1.2mmol dimethoxy Cynuric Chloride (CDMT); 3mmol n-formyl sarcolysine base morpholine (NMO); After the stirring at room 1 hour, add 1.1mmol tetrahydroisoquinoline (THIQ), continue stirred overnight.Add 1N HCl aqueous solution cancellation reaction, dichloromethane extraction, anhydrous sodium sulfate drying, the ETHYLE ACETATE column chromatography is received glassy compound, is primary amine shown in the formula Iia3 (being specially compound shown in the 4-1 in the above-mentioned reaction equation), productive rate 90%.
2) with compound dissolution shown in the 4mmol step 1) gained 4-1 in 4 milliliters of dry DMF, add 1 milliliter of thionyl chloride.0 ℃ was stirred 1 hour down, disappears according to TLC monitoring reaction raw material.Reaction solution is through ethyl acetate extraction and use the ETHYLE ACETATE silica gel column chromatography, obtains compound shown in the formula IIb3 (being specially compound 4-2 in the above-mentioned reaction equation), is white solid, and productive rate is 78%.
3) with 0.5mmol step 2) compound dissolution shown in the gained 4-2 in 5 milliliters of methylene dichloride, add 10 milliliters of methyl alcohol, freezingly fed the dry salt acid gas while stirring 1 hour to-20 ℃, measure and be 1L, and-20 ℃ of static spending the night down.Reaction liquid splashes in 0 ℃ of refrigerative 6N aqueous hydrochloric acid.Using sodium hydrogencarbonate adjustment pH is 7, ethyl acetate extraction.Anhydrous sodium sulfate drying.Silica gel column chromatography obtains compound shown in the formula IIc3 (being specially compound 4-3 in the above-mentioned reaction equation), productive rate 58%.
4) with compound dissolution shown in the 0.1mmol step 3) gained 4-3 in 4 milliliters of THFs; Add the Li-Al hydrogen of 10mmol, reflux after 24 hours, react with the 2N sodium-hydroxide treatment; Ethyl acetate extraction; Pass through the conventional column chromatography of ODS then, moving phase is methyl alcohol: water=1: obtain compound shown in the formula IId4 (being specially compound 4 in the reaction equation), productive rate 81% 1..
Compound 4:oil, 1HNMR (D 2O/300MHz) δ 7.11-6.99 (m, 4H), 3.63-3.57 (m, 3H), 3.63-3.33 (m, 1H), 2.84-2.72 (m, 3H), 2.69-2.37 (m, 2H), 2.28 (s, 3H), 2.10-2.02 (m, 1H), 1.95-1.79 (m, 1H), 1.51-1.38 (m, 2H); 13CNMR (75MHz/CDCl 3) δ 134.9,134.4,128.7,126.5,126.1,125.6,67.7,64.7,63.5,61.1,56.99,39.99,29.8,29.1,26.1; IR (KBr) 3411,2934,1641,1450,1050cm -1MS (ESI) m/z 261.4 (M ++ 1,100%) Anal.Calcd.for [M+H] C, 16H 25N 2O, 261.1967, Found: [M+H] 261.1961.
5) step 4) gained compound 4 is taken out 1mmol, be dissolved in the 10mL methylene dichloride, add the 10mmol diacetyl oxide, the 1mmol pyridine, the 0.1mmol 4-dimethylaminopyridine stirred 12 hours at 25 ℃, and reaction finishes and obtains pseudo-nucleoside compound 4 ' shown in the formula I.
Compound 4 ': o7.21-6.95 (m, 4H), 3.65-3.52 (m, 3H), 3.67-3.31 (m, 1H), 2.85-2.70 (m, 3H), 2.67-2.33 (m, 2H), 2.28 (s, 3H), 2.10-2.02 (m, 4H), 1.95-1.79 (m, 1H), 1.51-1.38 (m, 2H); 13CNMR (75MHz/CDCl 3) δ 134.5,134.7,128.3,126.4,126.1,122.6,67.7,62.7,61.5,60.1,56.4,39.5,29.7,28.1,22.1,21.9; IR (KBr) 3411,2934,1641,1450,1050cm -1MS (ESI) m/z 261.4 (M ++ 1,100%) Anal.Calcd.for [M+H] C, 18H 26N 2O 2, 303.2073, Found: [M+H] 303.2070.
By on can know that this compound structure is correct, be compound 4 ' shown in the formula I, wherein, n is 1, R 2For-CH 3, m is 1, Y does
Figure BDA0000056120000000211
Wherein, R 4Be H, X is-OCOCH 3
Embodiment 5: the compound 5,6 and 7 shown in the preparation formula III
Its reaction formula is following:
Figure BDA0000056120000000212
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; Disposable then adding is dissolved in substrate acid amides shown in the formula V1 of 1 mmole (247 milligrams) in 2.5 milliliters of DMSO 99.8MIN.s, puts into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Entire reaction is monitored with performance liquid, reacts stopped reaction after 39 hours, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) lyophilize (50 to-60 ℃) of step 1) gained filtrating is soaked into residue with 2 ml methanol, and the diethyl ether solution that splashes into diazomethane again carried out esterification reaction of organic acid 12 hours under-15 ℃ condition; Finish the back and add less water, utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying; Utilize 100-200 purpose silica gel column chromatography after concentrating, elutriant is ETHYLE ACETATE-ETHYLE ACETATE: methyl alcohol=20: 1 obtains diamide compound shown in the formula III provided by the invention (being specially the compound 5 in the above-mentioned reaction formula) (productive rate: 49%; Ee＞99.5%; Dicel-ADH), monoamide carboxylate methyl ester (being specially 6 in the above-mentioned reaction formula) (productive rate: 39%, ee＞99.5%, Dicel-ADH) with dicarboxylicacid methyl esters (being specially the compound 7 in the above-mentioned reaction formula) (productive rate: 6%; Ee＞99.5%, Dicel-ODH).
5：solid?mp?230℃；[α] D 25＝-38.0(c2.0，CH 3OH)； 1HNMR(DMSO-d6/300MHz)δ7.32-723(m，5H)，7.16(br?s，2H，CONH)，6.87(br?s，2H，CONH)，3.81(d，1H，J＝13.4Hz)，3.69(d，1H，J＝13.4Hz)，3.56-3.54(m，2H)，2.23-2.09(m，2H)，1.71-1.61(m，2H)； 13CNMR(75MHz/DMSO-d6)δ175.9，139.2，128.5，128.1，126.8，64.0，53.1，28.9；IR(KBr)3417(CONH 2)，1670，1628cm -1；MS(ESI)m/z?248.2(M ++1，100％)，270.2(M ++23，92.1).Anal.Calcd.for?C 13H 17N 3O 2：C，63.14；H，6.93；N，16.99.Found：C，62.94；H，6.92；N，17.05.
6：solid?mp?140℃；[α] D 25＝+121.5(c4.0，CHCl 3)； 1HNMR(CDCl 3/300MHz)δ7.35-7.21(m，5H)，6.91(br?s，1H，CONH)，5.50(br?s，1H，CONH)，3.92(d，1H，J＝13.1Hz)，3.84(d，1H，J＝13.1Hz)，3.80-3.72(m，2H)，3.69(s，3H)，2.58-2.51(m，1H)，2.09-2.03(m，1H)，2.00-1.85(m，2H)； 13CNMR(75MHz/CDCl 3)δ177.7，173.5，138.0，128.74，128.66，127.6，65.9，62.9，54.3，51.4，29.4，28.6；IR(KBr)3446(CONH 2)，1734，1685，1658cm -1；MS(ESI)m/z?263.2(M ++1，100％)，285.2(M ++23，64.3)，301.2(M ++39，4.9).Anal.Calcd.for?C 14H 18N 2O 3：C，64.10；H，6.92；N，10.68.Found：C，63.85；H，6.90；N，10.94.
7：oil；[α] D 25＝+10.7(c3.0，C 6H 6)； 1HNMR(CDCl 3/300MHz)δ7.97(br?s，1H，CONH)，7.33-7.25(m，5H)，5.57(br?s，1H，CONH)，3.90(d，1H，J＝13.2Hz)，3.81(d，1H，J＝13.2Hz)，3.63-3.51(m，2H)，3.58(s，3H)，2.20-2.14(m，1H)，2.13-2.04(m，1H)，1.89-1.87(m，2H)； 13CNMR(75MHz/CDCl 3)δ177.9，175.7，137.2，129.3，128.5，127.6，67.7，66.2，59.0，52.0，30.7，30.4；IR(KBr)3415(CONH 2)，1736，1679cm -1；MS(ESI)m/z?263.2(M ++1，59.2)，285.2(M ++23，100％)，301.2(M ++39，21.2).Anal.Calcd.for?C 14H 18N 2O 3：C，64.10；H，6.92；N，10.68.Found：C，64.25；H，6.87；N，10.92.
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula III.
Embodiment 6: the compound 8,9 and 10 shown in the preparation formula III
Its reaction formula is following:
Figure BDA0000056120000000221
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; Disposable then adding is dissolved in the formula V2 substrate acid amides of 1 mmole (197 milligrams) in 2.5 milliliters of DMSO 99.8MIN.s, puts into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Entire reaction is monitored with performance liquid, reacts stopped reaction after 7 hours, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, residue is soaked into 2 ml methanol, the diethyl ether solution that splashes into concentration again and be the diazomethane of 0.5mol/L carried out esterification reaction of organic acid 12 hours under-15 ℃ condition; Finish the back and add less water; Utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying, utilize 100-200 purpose silica gel column chromatography after concentrating; Elutriant is ETHYLE ACETATE-ETHYLE ACETATE: methyl alcohol=20: 1; Obtain diamide compound shown in the formula III provided by the invention (being specially the compound 8 in the above-mentioned reaction formula) (productive rate: 45%, ee＞99.5%, Dicel-ADH), monoamide carboxylate methyl ester (being specially the compound 9 in the above-mentioned reaction formula) (productive rate 36%; Ee＞96.8%; Dicel-ADH) and dicarboxylicacid methyl esters (being specially the compound compound 10 in the above-mentioned reaction formula) (productive rate: 10%, ee 93.3%, Dicel-OJH).
8：mp?222℃；[α] D 25＝-54.7(c1.5，CH 3OH)； 1HNMR(DMSO-d6/300MHz)δ7.23(br?s，2H，CONH)，6.87(br?s，2H，CONH)，5.95-5.84(m，1H)，5.15-5.01(m，2H)，3.55-3.53(m，2H)，3.28(dd，1H，J＝13.6，7.2Hz)，3.15(dd，1H，J＝13.5，5.7Hz)，2.18-2.07(m，2H)，1.69-1.59(m，2H)； 13CNMR(75MHz/DMSO-d6)δ176.0，136.4，116.5，64.3，52.5，28.9；IR(KBr)3415(CONH 2)，3381(CONH 2)，3200(CONH 2)，1660，1639cm -1；MS(ESI)m/z?198.1(M ++1，100％)，220.2(M ++23，17.1).Anal.Calcd.for?C 9H 15N 3O 2：C，54.81；H，7.67；N，21.30.Found：C，54.75；H，7.71；N，21.36.
9：mp?104℃；[α] D 25＝+78.0(c1.0，CHCl 3)； 1HNMR(CDCl 3/300MHz)δ6.91(br?s，1H，CONH)，5.89-5.78(br?s+m，1CONH+1H)，5.20-5.10(m，2H)，3.93(d，1H，J＝7.5Hz)，3.69(s，3H)，3.61(dd，1H，J＝10.9，3.3Hz)，3.36-3.34(m，2H)，2.54-2.47(m，1H)，2.15-2.12(m，1H)，1.96-1.87(m，2H)； 13CNMR(75MHz/CDCl 3)δ178.2，173.5，134.8，118.0，65.4，63.3，53.1，51.4，29.3，28.6；IR(KBr)3377(CONH 2)，3190(CONH 2)，1728，1654cm -1；MS(ESI)m/z?213.2(M ++1，100％)，235.2(M ++23，6.5).Anal.Calcd.for?C 10H 16N 2O 3：C，56.59；H，7.60；N，13.20.Found：C，56.57；H，7.61；N，12.94.
10：oil；[α] D 25＝+72.0(c1.5，CHCl 3)； 1HNMR(CDCl 3/300MHz)δ5.90-5.88(m，1H)，5.18-5.05(m，2H)，3.89-3.85(m，2H)，3.70(s，6H)，3.40-3.33(m，2H)，2.34-2.28(m，2H)，1.93-1.92(m，2H)； 13CNMR(75MHz/CDCl 3)δ174.7，135.3，117.7，63.6，53.5，51.6，28.4；IR(KBr)1738cm -1；MS(EI)m/z?227(M +，3％)，169(10)，168(100％)，108(11)；Anal.Calcd.for?C 11H 17NO 4：227.1158(M).Found：227.1161.
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula III.
Embodiment 7: the compound 5 and 11 shown in the preparation formula III
Its reaction formula is following:
Figure BDA0000056120000000241
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; The disposable then formula V3 substrate acid amides that adds 1 mmole (157 milligrams) is put into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Stopped reaction after 1 hour is reacted in entire reaction TLC monitoring, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, with residue with 2 milliliters of N, N '-dimethyl amide; 1 milliliter of benzyl bromine, 1.38 gram salt of wormwood stirring at room were carried out esterification in 24 hours and are finished back adding less water, utilized ethyl acetate extraction then; Behind the anhydrous sodium sulfate drying; Utilize 100-200 purpose silica gel column chromatography after concentrating, elutriant is ETHYLE ACETATE-ETHYLE ACETATE: methyl alcohol=20: 1 obtains diamide compound shown in the formula III provided by the invention (being specially the compound 5 in the above-mentioned reaction formula) (productive rate: 45%; Ee＞99.5%; Dicel-ADH) and monoamide benzyl carboxylate (being specially the compound 11 in the above-mentioned reaction formula) (productive rate: 36%, ee＞99.5%, Dicel-ADH).
11：solid；104.0℃；IR(KBr)v?3396，3172(CONH 2)，1726，1657，cm -1； 1H?NMR(300MHz，CDCl 3)(CDCl 3/300MHz)δ7.40-7.10(m，10H)，6.95(br?s，1H)，5.48(br?s，1H)，5.19(d，1H，J＝6.1Hz)，5.08(d，1H，J＝6.1Hz)，3.91-3.71(m，4H)，2.55-2.50(m，1H)，2.01-1.85(m，3H)； 13C?NMR(75MHz?CDCl 3)δ177.9，172.9，137.9，135.6，128.9，128.6，128.5，127.5，66.3，65.8，62.9，54.1，29.4，28.6；MS(ESI)m/z?339.1(M ++1，100％)，361.2(M ++23，82％).Anal.Calcd.for?C 20H 22N 2O 3：C，70.99；H，6.55；N，8.28；Found：C，70.66；H，6.62；N，8.43。
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula III.
Compounds X shown in embodiment 8, the preparation formula III "
Its reaction formula is following:
Figure BDA0000056120000000242
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; Disposable then adding is dissolved in substrate acid amides shown in the formula V4 of 1 mmole (291 milligrams) in 2.5 milliliters of DMSO 99.8MIN.s, puts into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Entire reaction is monitored with performance liquid, reacts stopped reaction after 39 hours, obtains containing the reaction solution of compound shown in the product formula X, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) the step 1) gained is contained the filtrating lyophilize (50 to-60 ℃) of compound shown in the formula X, residue is soaked into 2 ml methanol, the diethyl ether solution that splashes into diazomethane again carried out esterification reaction of organic acid 12 hours under-15 ℃ condition; Finish the back and add less water, utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying; Utilize 100-200 purpose silica gel column chromatography after concentrating; Elutriant is ETHYLE ACETATE-ETHYLE ACETATE: methyl alcohol=20: 1 gets the monoamide carboxylate methyl ester and (is specially the X ") in the above-mentioned reaction formula, productive rate: 39%; ee＞99.5%, Dicel-ADH.
X：solid?mp?150℃； 1HNMR(DMSO-d6/300MHz)δ14.55(br，s，1H)，7.58-7.30(m，7H)，5.10-5.0(m，2H)，4.90-4.45(m，2H)，4.21-4.15(m，2H)，2.46-2.38(m，2H)，1.95-1.73(m，2H)； 13CNMR(75MHz/DMSO-d6)δ175.3，175.1，174.9，174.7，153.6，153.4，136.4，136.4，128.3，127.8，127.1，126.99，65.51，66.46，61.5，61.0，60.5，59.99，30.2，29.3，28.2.；IR(KBr)3373，2361，1712，1644，1590，1411，1352，1229cm -1；MS(ESI)m/z?293.34(M ++1，10％)，315.29(M ++23，100％).Anal.Calcd.for：C 14H 16N 2O 5?C，57.53；H，5.52；N，9.58；Found：C，57.75；H，5.51，N，9.58.
X“：oil； 1HNMR(CDCl 3/300MHz)δ7.30-7.18(m，7H)，5.00-4.80(m，2H)，4.40-4.30(m，1H)，4.01-3.95(m，1H)，3.74(s，1.5H)，3.65(s，1.5H)，2.52-2.20(m，2H)，1.95-1.66(m，2H)； 13CNMR(75MHz/CDCl3)δ176.0，175.8，175.1，173.5，153.4，152.1，136.9，136.5，128.2，128.1，128.0，127.5，127.3，68.2，68.1，61.5，61.1，60.5，59.1，52.1，52.0，29.6，29.4，29.1，28.8；IR(KBr)3432，3401，1688，1412，1355，1212，1116，1004cm -1；MS(ESI)m/z?307.35(M ++1，20％)，329.29(M ++23，100％).Anal.Calcd.for：C 15H 18N 2O 5?C，58.82；H，5.92；N，9.15?Found：C，58.87；H，5.99；N.9.10.
By on can know that the above-claimed cpd structure is correct, for compounds X shown in the formula III ".
Embodiment 9: the compound 12 and 13 shown in the preparation formula II
Its reaction formula is following:
Figure BDA0000056120000000251
The practical implementation method:
1) get 2 the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml～250ml) washes thalline in threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; The disposable then formula IV1 substrate acid amides that adds 1 mmole～200 mmoles (157 milligrams～30 gram) is put into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Stopped reaction after 6 hours is reacted in entire reaction TLC monitoring, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, the spent ion exchange resin purifying obtains yellow solid, methyl alcohol flush away yellow impurities get monoamide monocarboxylic acid 12 (productive rate: 92～96%, ee 96%, Dicel-ADH).Again with residue with 2 milliliters of N, N '-N, 1 milliliter of benzyl bromine; Carried out in 24 hours adding less water after esterification finishes 1.38 gram salt of wormwood room temperature stirs for 25 ℃, utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying; Utilize 100-200 purpose silica gel column chromatography after concentrating, elutriant is an ETHYLE ACETATE, obtains the benzyl carboxylate of compound monoamide shown in the formula II provided by the invention (being specially the compound 13 in the above-mentioned reaction formula) (productive rate: 96%; Ee 96%, Dicel-ADH).
12：solid；mp?233-234℃?IR(KBr)v?3308，3153，1696，1642，1447，1403，1330，1308cm-1； 1HNMR(D2O/300MHz)δ4.34(t，J＝7.2Hz，1H)，4.11(t，J＝6.3Hz，1H)，2.39-2.25(m，2H)，2.05-1.91(m，2H)； 13NMR(75MHz/D2O)δ173.8，171.4，61.8，60.3，29.8，29.2.MS(ESI)m/z159.35(M ++1，7％)，181.46(M ++23，100％)；Anal.Calcd.for?C 6H 10N 2O 3：C，45.57；H，6.37；N，17.71；Found：C，45.54；H，6.53；N，17.78.
13：solid；mp?58.0℃；IR(KBr)v?3422，3369(CONH 2)，1740，1663cm -1； 1H?NMR(300MHz，CDCl3)δ7.78(br?s，1H)，7.28-7.14(m，10H)，6.17(br?s，1H)，4.98(s，2H)，3.89(d，1H，J＝6.6Hz)，3.79(d，1H，J＝6.6Hz)，3.63(t，1H，J＝7.3Hz)，3.52(dd，1H，J＝16.2Hz，4.3Hz)2.19-1.88(m，4H)； 13C?NMR(75MHz，CDCl 3)δ177.1，174.1，136.4，134.7，128.5，127.8，127.6，127.3，126.8，67.1，65.9，65.5，58.2，29.8，29.5；MS(ESI)m/z(％)39.1(M ++1，86％)361.2(M ++23，100％)；Anal.Calcd.for?C 20H 22N 2O 3：C，70.99；H，6.55；N，8.28；Found：C，70.65；H，6.71；N，8.53.
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula II.
Embodiment 10: the compound 14 shown in the preparation formula II
Its reaction formula is following:
Figure BDA0000056120000000261
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; The disposable then formula IV2 substrate acid amides that adds 1 mmole (171 milligrams) that is dissolved in 2.5 milliliters of DMSO 99.8MIN.s is put into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Stopped reaction after 60 hours is reacted in entire reaction TLC monitoring, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, residue is soaked into 2 ml methanol, the diethyl ether solution that splashes into concentration again and be the diazomethane of 0.5mol/L carried out esterification reaction of organic acid 12 hours under-15 ℃ condition; Finish the back and add less water; Utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying, utilize 100-200 purpose silica gel column chromatography after concentrating; Elutriant is an ETHYLE ACETATE; Obtain the carboxylate methyl ester of compound monoamide shown in the formula II provided by the invention (being specially the compound 14 in the above-mentioned reaction formula) (productive rate: 80%, ee 60.0%, Dicel-OJH).
14：solid，mp?93.0℃?IR(KBr)v?3419，(CONH 2)，1732，1683cm -1； 1H?NMR(300MHz，CDCl 3)δ7.62(br，1H)，5.64(br，1H)，3.68(s，3H)，3.33-3.28(m，1H)，3.13(dd，1H，J＝18.0，4.9Hz)，2.41(s，3H)，2.18-2.07(m，2H)，1.95-1.81(m，2H)； 13C?NMR(75MHz，CDCl 3)δ177.5，174.6，69.6，68.2，52.1，40.9，30.3，29.9；MS(ESI)m/z(％)187.1(M ++1，14％)，209.1(M ++23，100％).；Anal.Calcd.for?C 8H 14N 2O 3：C，51.60；H，7.58；N，15.04；Found：C，51.14；H，7.40；N，15.38.。
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula II.
Embodiment 11: the compound 15 shown in the preparation formula II
Its reaction formula is following:
Figure BDA0000056120000000271
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; The disposable then formula IV3 substrate acid amides that adds 1 mmole (197 milligrams) is put into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Stopped reaction after 7 days is reacted in entire reaction TLC monitoring, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, residue is soaked into 2 ml methanol, the diethyl ether solution that splashes into concentration again and be the diazomethane of 0.5mol/L carried out esterification reaction of organic acid 12 hours under-15 ℃ condition; Finish the back and add less water; Utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying, utilize 100-200 purpose silica gel column chromatography after concentrating; Elutriant is an ETHYLE ACETATE; Obtain the carboxylate methyl ester of compound monoamide shown in the formula II provided by the invention (being specially the compound 15 in the above-mentioned reaction formula) (productive rate: 60%, ee 95.2%, Dicel-ADH).
15：solid；mp?72.0-74.0℃；IR(KBr)v?3364，3292，3265，1686，1650cm -1；[α] 25 D-64°(c?0.5，CHCl 3)； 1H?NMR(300MHz，CDCl 3)δ7.43(s，1H)，7.37-7.42(m，1H)，7.18-7.20(m，2H)，7.03(s，1H)，5.58(s，1H)，3.73(d，J＝12.9Hz，1H)，3.69(t，J＝8.6Hz，1H)，3.51(d，J＝13.1Hz，1H)，3.34-3.39(m，1H)，2.98(q，J＝16.3Hz，1H)，2.38-2.48(m，1H)，2.13-2.25(m，1H)； 13C?NMR(75MHz，CDCl 3)δ179.5，139.7，131.4，130.5，130.1，127.1，122.6，66.1，61.6，50.9，23.0；MS(ESI)m/z(％)269[M+H] +，271[M+H+2] +；Anal.Calcd.for?C 11H 13BrN 2O：C，49.09；H，4.87；N，10.41.Found：C，49.10；H，4.68；N，10.23.
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula II.
Embodiment 12: the compound 16 shown in the preparation formula II
Its reaction formula is following
Figure BDA0000056120000000272
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; The disposable then formula IV4 substrate acid amides that adds 1 mmole (247 milligrams) that is dissolved in 2.5 milliliters of DMSO 99.8MIN.s is put into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Stopped reaction after 4 days is reacted in entire reaction TLC monitoring, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, residue is soaked into 2 ml methanol, the diethyl ether solution that splashes into concentration again and be the diazomethane of 0.5mol/L carried out esterification reaction of organic acid 12 hours under-15 ℃ condition; Finish the back and add less water; Utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying, utilize 100-200 purpose silica gel column chromatography after concentrating; Elutriant is an ETHYLE ACETATE; Obtain the carboxylate methyl ester of compound monoamide shown in the formula II provided by the invention (being specially the compound 16 in the above-mentioned reaction formula) (productive rate: 15%, ee＞99.5%, Dicel-ADH).
16：oil； 1HNMR(CDCl 3/300MHz)δ7.97(br?s，1H，CONH)，7.33-7.25(m，5H)，5.57(br?s，1H，CONH)，3.90(d，1H，J＝13.2Hz)，3.81(d，1H，J＝13.2Hz)，3.63-3.65(m，2H)，3.58(s，3H)，2.20-2.14(m，1H)，2.13-2.04(m，1H)，1.89-1.87(m，2H)； 13CNMR(75MHz/CDCl3)δ177.9，175.7，137.2，129.3，128.5，127.6，67.7，66.2，59.0，52.0，30.7，30.4；IR(KBr)3415(CONH2)，1736，1679cm -1；MS(ESI)m/z?263.2([M+23] +，100％)，301.2([M+39] +，21.2％)；Anal.Calcd.ForC 14H 18N 2O 3：C，64.10；H，6.92；N，10.68.Found：C，64.25；H，6.87；N，10.92.
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula II.
Embodiment 13: the compound 17 shown in the preparation formula II
Its reaction formula is following:
Figure BDA0000056120000000281
The practical implementation method is:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline; Thawed 30 minutes under 30 ℃ of conditions, with the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0; 50ml) thalline is washed in the Erlenmeyer Florence flask of 150 milliliters of threaded mouths; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes, disposable then adding 1 mmole (171 milligrams) 2,6-two replaces six hydrogen piperidines acid amides; Put into 30 ℃ of shaking tables, carry out catalytic hydrolysis reaction under the 200rpm condition.Entire reaction is monitored with TLC, reacts stopped reaction after 1 hour, and the gained reaction solution is gone out thalline through one deck zeyssatite suction filtration, and water respectively washs filter residue three times for 15 milliliters.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, with residue with 2 milliliters of N, N '-N; 1 milliliter of benzyl bromine, 1.38 gram salt of wormwood room temperatures stir for 25 ℃ and carried out in 24 hours adding less water after esterification finishes, and utilize ethyl acetate extraction then; Behind the anhydrous sodium sulfate drying; Utilize 100-200 purpose silica gel column chromatography after concentrating, elutriant is an ETHYLE ACETATE, obtains the benzyl carboxylate of compound monoamide shown in the formula II provided by the invention (being specially the compound 17 in the above-mentioned reaction formula) (productive rate: 96%; Ee＞99.5%, Dicel-ADH).
17：solid?mp?105.0℃?IR(KBr)v?3420(CONH 2)，1744，1632cm -1； 1H?NMR(300MHz，CDCl 3)δ7.36-7.17(m，11H)，5.68(br?d，1H，J＝22.4Hz)，5.07(t，2H，J＝12.6Hz)，3.76(s，2H)，3.23(dd，1H，J＝21.03，3.78Hz，)，3.09(dd，1H，J＝18.8，4.0Hz)，2.00-1.92(m，2H)1.89-1.62(m，3H)1.26-1.17(m，1H)； 13C?NMR(75MHz，CDCl 3)δ177.3，174.0，135.6，134.9，130.3，128.6，128.5，128.4，128.2，127.6，66.7，63.3，61.6，58.3；MS(ESI)m/z(％)353.2(M ++1，100％)，375.2(M ++23，20％).；Anal.Calcd.for?C 21H 24N 2O 3，C，71.57；H，6.86；N，7.95；Found：C，71.58；H，6.84，N，7.96..
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula II.
Embodiment 14: the compound 18 shown in the preparation formula II
Its reaction formula is following:
Figure BDA0000056120000000291
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline; Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; The formula IV5 acid amides of 1 mmole of disposable adding (185 milligrams) is put into 30 ℃ of shaking tables then, carries out catalytic hydrolysis reaction under the 200rpm condition.Entire reaction is monitored with TLC, reacts stopped reaction after 1 hour, and the gained reaction solution is gone out thalline through one deck zeyssatite suction filtration, and water respectively washs filter residue three times for 15 milliliters.
2) with the lyophilize (50 to-60 ℃) of step 1) gained filtrating, with residue with 2 milliliters of N, N '-N; 1 milliliter of benzyl bromine; Carried out in 24 hours adding less water after esterification finishes 1.38 gram salt of wormwood room temperature stirs for 25 ℃, utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying; Utilize 100-200 purpose silica gel column chromatography after concentrating; Elutriant is an ETHYLE ACETATE, obtains the benzyl carboxylate of compound monoamide shown in the formula II provided by the invention (being specially the compound 18 in the above-mentioned reaction formula) (productive rate: 74%, ee 67%).
18：oil?IR(KBr)v?3410(CONH 2)，1737，1670cm -1； 1H?NMR(300MHz，CDCl 3)δ8.41(br，1H)，3.38-7.22(m，10H)，5.95(br，1H)，5.18-5.04(dd，J＝2H)，3.74-3.63(dd.2H)，3.61-3.60(1H)，3.44-3.40(dd，1H)，2.02-1.46(m，8H)； 13C?NMR(75MHz，CDCl 3)δ178.26，176.33，137.96，135.45，128.95，128.67，128.537，128.51，128.30，127.62，67.04，60.632，29.13，28.20，28.13，26.80；MS(ESI)m/z(％)367.18(M ++1，20％)，389.26(M ++23，100％)；Anal.Calcd.for?C 22H 26N 2O 3?C，72.11；H，7.15；N，7.64；Found：C，72.07；H，7.10；N，7.79.
By on can know that the above-claimed cpd structure is correct, be compound shown in the formula II.
Embodiment 15: the compounds X shown in the preparation formula II '
Its reaction formula is following:
Figure BDA0000056120000000292
The practical implementation method:
1) get two the gram weight in wet bases Rhodococcus erythropolis AJ270 thalline (Institute of Chemistry, Academia Sinica); Thawed 30 minutes under 30 ℃ of conditions; With the buffered soln of potassium hydrogenphosphate and potassium primary phosphate (0.1M, pH 7.0,50ml) thalline washed in 150 milliliters threaded mouthful the Erlenmeyer Florence flask; Put under 30 ℃ of conditions of shaking table activation after disperseing to shake up 30 minutes; Disposable then adding is dissolved in substrate acid amides shown in the formula IV6 of 1 mmole (291 milligrams) in 2.5 milliliters of DMSO 99.8MIN.s, puts into 30 ℃ of shaking tables, carries out catalytic hydrolysis reaction under the 200rpm condition.Entire reaction is monitored with performance liquid, reacts stopped reaction after 39 hours, obtains containing the reaction solution of compound shown in the product formula X, and the gained reaction solution is removed thalline through one deck zeyssatite suction filtration, and 20 milliliters of washings of water filter residue is three times successively.
2) step 1) gained filtrating lyophilize (50 to-60 ℃) is soaked into residue with 2 ml methanol, the diethyl ether solution that splashes into concentration again and be the diazomethane of 0.5mol/L carried out esterification 12 hours under-15 ℃ condition; Finish the back and add less water; Utilize ethyl acetate extraction then, behind the anhydrous sodium sulfate drying, utilize 100-200 purpose silica gel column chromatography after concentrating; Elutriant is an ETHYLE ACETATE; Monoamide carboxylate methyl ester (being specially the x ' in the above-mentioned reaction formula) (productive rate: 39%, ee＞99.5%, Dicel-ADH).
x：solid?mp?198℃； 1HNMR(DMSO-d6/300MHz)δ13.55(br，s，1H)，7.88-7.33(m，7H)，5.15-5.03(m，2H)，4.94-4.90(m，2H)，4.41-4.23(m，2H)，2.36-2.28(m，2H)，1.85-1.77(m，2H)； 13CNMR(75MHz/DMSO-d6)δ175.3，175.1，174.9，174.7，153.6，153.4，136.4，136.4，128.3，127.8，127.1，126.99，65.51，66.46，61.5，61.0，60.5，59.99，30.2，29.3，28.2.；IR(KBr)3373，2361，1712，1644，1590，1411，1352，1229cm -1；MS(ESI)m/z293.34(M++1，10％)，315.29(M ++23，100％).Anal.Calcd.for：C 14H 16N 2O 5?C，57.53；H，5.52；N，9.58；Found：C，57.75；H，5.51，N，9.58.
x’：solid?mp?104℃； 1HNMR(CDCl 3/300MHz)δ7.34-7.20(m，7H)，5.07-4.90(m，2H)，4.40-4.31(m，1H)，4.11-4.05(m，1H)，3.64(s，1.5H)，3.55(s，1.5H)，2.42-2.10(m，2H)，1.85-1.76(m，2H)；13CNMR(75MHz/CDCl3)δ175.0，174.8，174.3，174.0，154.4，154.1，136.9，136.7，128.8，128.4，128.3，127.7，127.6，67.2，67.1，62.5，62.1，60.5，60.1，53.1，53.0，30.6，29.7，29.6，28.8；IR(KBr)3432，3401，1688，1412，1355，1212，1116，1004cm-1；MS(ESI)m/z307.35(M ++1，20％)，329.29(M ++23，100％).Anal.Calcd.for：C 15H 18N 2O 5?C，58.82；H，5.92；N，9.15?Found：C，58.87；H，5.99；N，9.10.
By on can know that the above-claimed cpd structure is correct, for compounds X shown in the formula II '.
Embodiment 16: the anti-tumor activity of pseudo-nucleoside compound shown in the formula I
Tetrazole [MTT, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] be a kind of dyestuff that can accept Wasserstoffatoms.Desaturase relevant with NADP in the viable cell plastosome can change into insoluble hepatic formazon with xanchromatic MTT in cell, dead cell does not then have this function.Behind DMSO dissolving formazon, under certain wavelength, measure OD value with ELIASA, both can quantitatively measure the survival rate of cell.Experimental technique is following: select the adherent tumour cell of logarithmic phase for use, after trysinization, be made into the cell suspension of 5000/ml with the RPMI1640 nutrient solution of 10% calf serum; Be seeded in 96 well culture plates; Every hole inoculation 100ul, 37 ℃, 5%CO2 cultivates 24h.
Experimental group adds sample 10ul, and every hole final volume is 200ul, supplies with RPMI-1640.37 ℃, 5%CO 2Cultivate 3d.
Abandon supernatant, every hole adds the serum-free medium of the freshly prepared 0.5mg/ml MTT of 100ul, and 37 ℃ are continued to cultivate 4h.Carefully abandon supernatant, and add 200ulDMSO dissolving MTT formazon deposition,, on ELIASA, measure the OD value at wavelength 544nm place with miniature ultrasonic vibrator mixing.
Growth of tumour cell inhibiting rate=(OD Contrast-OD Experiment)/(OD Contrast-OD Blank) * 100%
The growth inhibition ratio that has recorded the embodiment 1 preparation pseudo-1 couple of Bel-7402 of nucleoside compound of gained (liver cancer cell) is 1.96%, and the growth inhibition ratio of A-549 (lung carcinoma cell) is 1.96%.By on can know that the growth of pseudo-1 couple of Bel-7402 of nucleoside compound (liver cancer cell) of the embodiment of the invention 1 preparation gained and A-549 (lung carcinoma cell) all has certain restraining effect.

Claims (18)

1. pseudo-nucleoside compound shown in the formula I;
Figure FDA0000056119990000011
(formula I)
Among the said formula I, n is the integer of 1-3, and m is 0 or 1;
* represent chirality, be R or S configuration;
X is-OH,
Figure FDA0000056119990000012
Wherein, R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
Y is-OH,
Figure FDA0000056119990000013
Wherein, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN.
One kind prepare shown in the said formula I of claim 1 in m be 0, Y is the method for
Figure FDA0000056119990000014
X for the pseudo-nucleoside compound of-OH,
Figure FDA0000056119990000015
Figure FDA0000056119990000016
; Be method a or method b; Wherein, said method a comprises the steps:
1) with said compound of the said formula II ' of claim 12 and SOCl 2In N, to react in the dinethylformamide, reaction finishes and obtains compound shown in the formula IIa1;
Figure FDA0000056119990000017
(formula II ')
Among the said formula II ', n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
(formula IIa1)
2) with compound and NaN shown in the step 1) gained formula IIa1 3And ZnBr 2In the mixed solution of being made up of Virahol and water, carry out back flow reaction, reaction finishes and obtains compound shown in the formula IIb1;
Figure FDA0000056119990000022
(formula IIb1)
3) with step 2) compound and LiAlH shown in the gained formula IIb1 4In THF, carry out back flow reaction, reaction finishes and obtains compound shown in the formula IIc1;
(formula IIc1)
4) compound shown in the formula IIc1 of step 3) gained is reacted with diacetyl oxide, pyridine and 4-dimethylamino-pyridine in methylene dichloride; Reaction finish obtain that m among the said formula I is 0, Y is for the pseudo-nucleoside compound of
Figure FDA0000056119990000024
X for-OH,
Figure FDA0000056119990000025
, also is compound shown in the formula IId1;
Figure FDA0000056119990000026
(formula IId1)
Among said formula IIa1, formula IIb1, formula IIc1 and the formula IId1, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; X is-OH,
Figure FDA0000056119990000031
R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5;
Said method b comprises the steps:
1) with the said formula III of claim 16 ' said compound and SOCl 2In DMF, react, reaction finishes and obtains compound shown in the formula III a1;
Figure FDA0000056119990000032
(formula III ')
Said formula III ' in, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3, R 3' be-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
Figure FDA0000056119990000033
(formula III a1)
2) with compound and NaN shown in the step 1) gained formula III a1 3And ZnBr 2In the mixed solution of being made up of Virahol and water, carry out back flow reaction, reaction finishes and obtains compound shown in the formula III b1;
Figure FDA0000056119990000034
(formula III b1)
3) with step 2) compound and LiAlH shown in the gained formula III b1 4In THF, carry out back flow reaction, reaction finishes and obtains compound shown in the formula III c1;
Figure FDA0000056119990000035
(formula III c1)
4) with compound shown in the formula III c1 of step 3) gained in methylene dichloride with diacetyl oxide; Pyridine and 4-dimethylamino-pyridine react; Reaction finishes and obtains that m is 0 among the said formula I; Y is that
Figure FDA0000056119990000041
X is-OH; The pseudo-nucleoside compound of
Figure FDA0000056119990000042
; It also is compound shown in the formula III d1
Figure FDA0000056119990000043
(formula III d1)
Among said formula III a1, formula III b1, formula III c1 and the formula III d1, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; X is-OH,
Figure FDA0000056119990000044
R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5.
3. method according to claim 2 is characterized in that: in the step 1) of said method a, and the said compound of said formula II ', SOCl 2With N, the amount ratio of dinethylformamide is 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferred 10mmol: 1mL: 4mL; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour;
Said step 2) in, compound, NaN shown in the said formula IIa1 3, ZnBr 2With the amount ratio of the said mixed solution of forming by Virahol and water be 0.1-10.0mmol: 0.2-20mmol: 0.1-10mmol: 1-10mL, preferred 1mmol: 2mmol: 0.5mmol: 4.5mL; In the said mixed solution of being made up of Virahol and water, the volume ratio of Virahol and water is 1-10: 0.1-100, preferred 1: 2; In the said back flow reaction step, the time is 1-24 hour, preferred 16 hours;
In the said step 3), compound, LiAlH shown in the said formula IIb1 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 1mmol: 10mmol: 10mL; In the said back flow reaction step, the time is 6-48 hour, preferred 12 hours;
In the said step 4); The amount ratio of compound, diacetyl oxide, pyridine, 4-dimethylaminopyridine and methylene dichloride shown in the said formula IIc1 is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours;
In the step 1) of said method b, said formula III ' said compound, SOCl 2With N, the amount ratio of dinethylformamide is 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferred 10mmol: 1mL: 4mL; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour;
Said step 2) in, compound, NaN shown in the said formula III a1 3, ZnBr 2With the amount ratio of the said mixed solution of forming by Virahol and water be 0.1-10.0mmol: 0.2-20mmol: 0.1-10mmol: 1-10mL, preferred 1mmol: 2mmol: 0.5mmol: 4.5mL; In the said mixed solution of being made up of Virahol and water, the volume ratio of Virahol and water is 1-10: 0.1-100, preferred 1: 2; In the said back flow reaction step, the time is 1-24 hour, preferred 16 hours;
In the said step 3), compound, LiAlH shown in the said formula III b1 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 1mmol: 10mmol: 10mL; In the said back flow reaction step, the time is 6-48 hour, preferred 12 hours;
In the said step 4); The amount ratio of compound, diacetyl oxide, pyridine, 4-dimethylaminopyridine and methylene dichloride is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL shown in the said formula III c1, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours.
One kind prepare shown in the said formula I of claim 1 in m be 1, Y is for-OH or
Figure FDA0000056119990000051
X method for the pseudo-nucleoside compound of-OH,
Figure FDA0000056119990000052
, comprise the steps:
1) with said compound of the said formula II ' of claim 12 and LiAlH 4In THF, carry out back flow reaction, reaction finishes and obtains compound shown in the formula IIa2;
Figure FDA0000056119990000053
(formula II ')
Among said formula II ' and the formula IIa2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
Figure FDA0000056119990000061
(formula IIa2)
2) with compound and R shown in the said step 1) gained formula IIa2 4PhCH 2CH 2CHO and NaBCNH 3In methylene dichloride or methyl alcohol, react, reaction finishes and obtains compound shown in the formula IIb2; Said R 4PhCH 2CH 2Among the CHO, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
Figure FDA0000056119990000062
(formula IIb2)
Among the said formula IIb2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a; Ph is a phenyl;
3) with said step 2) aqueous solution of compound shown in the gained formula IIb2 and hydrochloric acid and formaldehyde carries out back flow reaction; Reaction finish obtain that m among the said formula I is 1, X is for the compound of
Figure FDA0000056119990000063
Y for-OH, also is compound shown in the formula IIc2;
Figure FDA0000056119990000064
(formula IIc2)
Among the said formula IIc2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
4) with compound shown in the formula IIc2 of step 3) gained in methylene dichloride with diacetyl oxide; Pyridine and 4-dimethylamino-pyridine react; Reaction finishes and obtains that m is 1 among the said formula I; Y is that
Figure FDA0000056119990000071
X is-OH; The pseudo-nucleoside compound of
Figure FDA0000056119990000072
; It also is compound shown in the formula IId2
(formula IId2)
Among the said formula IId2, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a; X is-OH,
Figure FDA0000056119990000074
Figure FDA0000056119990000075
R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5.
5. method according to claim 4 is characterized in that: in the said step 1), and the said compound of said formula II ', LiAlH 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 1mmol: 10mmol: 10mL; In the said back flow reaction step, the time is 6-48 hour, preferred 24 hours;
Said step 2) in, compound, R shown in the said formula IIa2 4PhCH 2CH 2CHO, NaBCNH 3With the amount ratio of methylene dichloride or methyl alcohol be 0.1-10mmol: 0.1-10mmol: 1-100mmol: 1-100mL, preferred 1mmol: 1.0mmol: 2mmol: 6mL; In the said reactions step, temperature is 0-50 ℃, and preferred 25 ℃, the time is 12-48 hour, preferred 24 hours;
In the said step 3), the amount ratio of the aqueous solution of compound, formaldehyde, hydrochloric acid and chloroform shown in the said formula IIb2 is 0.1-10mmol: 0.1-20mL: 1-40mL: 0-40mL, preferred 1mmol: 2.4mL: 4.8mL: 4.8mL; In the said back flow reaction step, the time is 1-48 hour, preferred 24 hours; The mass percentage concentration of said hydrochloric acid is 10-35%, preferred 35%; The mass percentage concentration of the aqueous solution of said formaldehyde is 10-35%, preferred 35%;
In the said step 4); The amount ratio of compound shown in the formula IIc2, diacetyl oxide, pyridine, 4-dimethylaminopyridine and methylene dichloride is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours.
6. one kind prepares m=1 in the claim 1; X is-OH,
Figure FDA0000056119990000081
Y be the method for the pseudo-nucleoside compound of-OH or
Figure FDA0000056119990000082
, comprise the steps:
1) the said compound of the said formula II ' of claim 12 and dimethoxy Cynuric Chloride, nitrogen methylmorpholine and tetrahydroisoquinoline are reacted in methylene dichloride, reaction finishes and obtains compound shown in the formula IIa3;
Figure FDA0000056119990000083
(formula II ')
Among said formula II ' and the formula IIa3, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a;
(formula IIa3)
2) with compound and SOCl shown in the step 1) gained formula IIa3 2In DMF, react, reaction finishes and obtains compound shown in the formula IIb3;
Figure FDA0000056119990000085
(formula IIb3)
3) with step 2) compound shown in the gained formula IIb3 and the mixed solution of forming by methyl alcohol and methylene dichloride, feed dry HCl gas, react, reacting finishes obtains compound shown in the formula IIc3;
(formula IIc3)
4) with compound and LiAlH shown in the step 3) gained formula IIc3 4In THF, carry out back flow reaction, reaction finishes and obtains among the said formula I that m is 1, X does
Figure FDA0000056119990000092
Y is-compound of OH also is a compound shown in the formula IId3;
Figure FDA0000056119990000093
(formula IId3)
5) compound shown in the formula IId3 of step 3) gained is reacted with acetic anhydride, pyridine and 4- dimethylamino-pyridine in carrene; Reaction finishes and obtains m=1 among the said formula I; X is-OH, the pseudo- nucleoside compound of
Figure FDA0000056119990000094
Y for
Figure FDA0000056119990000095
; It also is compound shown in the formula IIe3
Figure FDA0000056119990000096
(formula IIe3)
Among the said formula IIb3-formula IIe3, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2Be-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; X is-OH,
Figure FDA0000056119990000101
R 1For the total number of carbon atoms is alkyl or the phosphate of 1-5, R 4Be selected from-H ,-F ,-Cl ,-Br ,-I ,-CH 3,-OCH 3,-NO 2With-HSO 3In at least a.
7. method according to claim 6; It is characterized in that: in the said step 1); The amount ratio of the said compound of said formula II ', dimethoxy Cynuric Chloride, nitrogen methylmorpholine, tetrahydroisoquinoline and methylene dichloride is 0.1-10mmol: 0.1-20mmol: 0.1-20mmol: 0.1-10mmol: 1-40mL, preferred 5mmol: 1.2mmol: 3mmol: 1.1mmol: 10mL; In the said reactions step, temperature is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours;
Said step 2) in, compound, SOCl shown in the said formula IIa3 2With the amount ratio of DMF be 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferred 4mmol: 1mL: 4mL; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour;
In the said step 3), compound shown in the said formula IIb3, the mixed solution amount ratio of being made up of methyl alcohol and methylene dichloride are 0.1-1mmol: 1-15mL, preferred 0.5mmol: 15mL; The feeding volume of said dry HCl gas is 22.4mL-22.4L, preferred 1L; In the said reactions step, temperature is-10 ℃-50 ℃, and preferred 0 ℃, the time is 0.5-24 hour, preferred 1 hour; In the said mixed solution of being made up of methyl alcohol and methylene dichloride, the volume ratio of methyl alcohol and methylene dichloride is 2: 1; In the said reactions step, temperature is-20-0 ℃, and preferred-20 ℃, the time is 0.5-12 hour, preferred 1 hour;
In the said step 4), compound, LiAlH shown in the said formula IIc3 4With the amount ratio of THF be 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferred 0.1mmol: 10mmol: 4mL; In the said back flow reaction step, the time is 6-48 hour, preferred 24 hours;
In the said step 5); The amount ratio of compound shown in the formula IId3, diacetyl oxide, pyridine, 4-dimethylaminopyridine and methylene dichloride is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferred 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; Said temperature of reaction is 0-50 ℃, and preferred 25 ℃, the time is 6-48 hour, preferred 12 hours.
8. be the antitumor drug of activeconstituents with pseudo-nucleoside compound shown in the said formula I of claim 1.
9. the application of pseudo-nucleoside compound shown in the said formula I of claim 1 in the preparation antitumor drug.
10. according to Claim 8 or 9 described application, it is characterized in that: said tumour is liver cancer or lung cancer.
11. compound shown in the formula II,
(formula II)
Among the said formula II, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
12. compound according to claim 11 is characterized in that: compound shown in the said formula II is a compound shown in the formula II ',
Figure FDA0000056119990000112
(formula II ')
Among the said formula II ', n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
13. method for preparing claim 11 or 12 said compounds; Be following method a or method b; Wherein, after said method a comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula IV, in solvent, carry out esterification with alkali and benzyl bromine or to the methoxybenzyl bromine again; Reaction finishes and obtains claim 11 or 12 said compounds
(formula IV)
Among the said formula IV, R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3;
After said method b comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula IV; Diethyl ether solution with diazomethane carries out esterification again; Reaction finishes and obtains claim 11 or 12 said compounds
Figure FDA0000056119990000121
(formula IV)
Among the said formula IV, R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3.
14. method according to claim 13 is characterized in that: among said method a and the method b, said rhodococcus catalystsystem is that the buffered soln of 6.0-8.0 is formed by rhodococcus and pH value; Said buffered soln is Na 2HPO 4-citric acid solution, K 2HPO 4-KH 2PO 4Buffered soln, Tris buffered soln, Hanks ' buffered soln or PBS buffered soln; The amount ratio of compound is 2g: 1mmol-2g: 1mol shown in said rhodococcus and the formula IV; In the said rhodococcus catalystsystem, the amount ratio of rhodococcus and said buffered soln is 2g: 50mL-2g: 1L; In the said catalytic hydrolysis reaction step, temperature is 20-37 ℃, and preferred 30 ℃, the time is 0.1-120 hour, preferred 7-39 hour;
Among the said method a, said alkali is salt of wormwood, yellow soda ash, Pottasium Hydroxide, sodium hydroxide or cesium carbonate, preferred salt of wormwood; Said solvent is selected from acetone, N, at least a in dinethylformamide, DMSO 99.8MIN. and the THF, preferred N, dinethylformamide; Said alkali, said solvent, benzyl bromine or be 1.38g: 2mL: 1mL: 1mmol to the amount ratio of compound shown in methoxybenzyl bromine and the said formula IV; In the said step of esterification, temperature is 10-30 ℃, and preferred 25 ℃, the time is 1-48 hour, preferred 24 hours;
Among the said method b, the concentration of the diethyl ether solution of said diazomethane is 0.1-1mol/L, preferred 0.5mol/L; In the said step of esterification, temperature is-20-10 ℃, and preferred 0 ℃, the time is 2-24 hour, preferred 12 hours.
15. compound shown in the formula III,
Figure FDA0000056119990000122
(formula III)
In the said formula III, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3, R 3' be-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
16. compound according to claim 15 is characterized in that: the said compound of said formula III is a formula III ' shown in compound,
Figure FDA0000056119990000131
(formula III ')
Said formula III ' in, n is the integer of 1-3; * represent chirality, be R or S configuration; R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN; R 3For-NH 2,-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3, R 3' be-OH ,-OCH 3,-OCH 2CH 3,-OCH 2CH=CH 2,-OCH 2C 6H 5Or-OCH 2C 6H 5OCH 3
17. method for preparing claim 15 or 16 said compounds; Be following method a or method b; Wherein, after said method a comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula V, in solvent, carry out esterification with alkali and benzyl bromine or to the methoxybenzyl bromine again; Reaction finishes and obtains claim 15 or 16 said compounds
Figure RE-FDA0000079357960000012
(formula V)
Among the said formula V, R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 3,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3;
After said method b comprises the steps: to carry out catalytic hydrolysis reaction with compound shown in rhodococcus (Rhodococcus erythropolis AJ270) catalystsystem and the formula V; Diethyl ether solution with diazomethane carries out esterification again; Reaction finishes and obtains claim 15 or 16 said compounds
Figure RE-FDA0000079357960000013
(formula V)
Among the said formula V, R 2For-H ,-CH 3,-CH 2CH 3,-CH 2CH=CH 2,-CH 2C 6H 5,-CH 2C 6H 5OCH 3,-Cbz ,-CH 2CH 2COCH 2,-CH 2CH 2COOCH 3,-CH 2CH 2CONH 2,-CH 2CH 2CHO or-CH 2CH 2CN, n are the integer of 1-3.
18. method according to claim 17 is characterized in that: among said method a and the method b, said rhodococcus catalystsystem is that the buffered soln of 6.0-8.0 is formed by rhodococcus and pH value; Said buffered soln is Na 2HPO 4-citric acid solution, K 2HPO 4-KH 2PO 4Buffered soln, Tris buffered soln, Hanks ' buffered soln or PBS buffered soln; The amount ratio of compound is 2g: 1mmol-2g: 1mol shown in said rhodococcus and the formula IV; In the said rhodococcus catalystsystem, the amount ratio of rhodococcus and said buffered soln is 2g: 50mL-2g: 1L; In the said catalytic hydrolysis reaction step, temperature is 20-37 ℃, and preferred 30 ℃, the time is 0.1-120 hour, preferred 7-39 hour;
Among the said method a, said alkali is salt of wormwood, yellow soda ash, Pottasium Hydroxide, sodium hydroxide or cesium carbonate, preferred salt of wormwood; Said solvent is selected from acetone, N, at least a in dinethylformamide, DMSO 99.8MIN. and the THF, preferred N, dinethylformamide; Said alkali, said solvent, benzyl bromine or be 1.38g: 2mL: 1mL: 1mmol to the amount ratio of compound shown in methoxybenzyl bromine and the said formula V; In the said step of esterification, temperature is 10-30 ℃, and preferred 25 ℃, the time is 1-48 hour, preferred 24 hours;
Among the said method b, the concentration of the diethyl ether solution of said diazomethane is 0.1-1mol/L, preferred 0.5mol/L; In the said step of esterification, temperature is-20-10 ℃, and preferred-15 ℃, the time is 2-24 hour, preferred 12 hours.
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CN114957030A (en) * 2021-02-24 2022-08-30 中国科学院化学研究所 Chiral cyclic compound and preparation method and application thereof

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JP2016102079A (en) * 2014-11-28 2016-06-02 株式会社豊田自動織機 Saturated heterocycle-containing compound, secondary battery electrode and secondary battery using the same, binder, and fluorescent substance
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CN110857276B (en) * 2018-08-22 2021-03-02 中国科学院化学研究所 Chiral beta-hydroxy amide compounds and preparation method and application thereof
CN114957030A (en) * 2021-02-24 2022-08-30 中国科学院化学研究所 Chiral cyclic compound and preparation method and application thereof
CN114957030B (en) * 2021-02-24 2023-09-22 中国科学院化学研究所 Chiral cyclic compound and preparation method and application thereof

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