CN102746278B - Chiral pseudonucleoside compound and its preparation method and application - Google Patents

Abstract

The invention discloses a chiral pseudo nucleoside compound and a preparation method and application thereof. The compound is shown as a formula I. The raw material for preparing the pseudo-nucleoside compound is obtained by catalyzing and hydrolyzing azacyclo-diamide compounds by using a Rhodococcus erythropolis AJ270 microbial system. The dosage of the rhodococcus thallus can be adjusted according to the dosage of the substrate. The reaction solvent is common buffer solution with pH value of 6.0-8.0, the temperature is 20-37 ℃, and the reaction time is 0.1-120 hours. The Rhodococcus microorganism catalysis system has the characteristics of fermentation culture and convenient preservation. The method for preparing chiral aza-cyclic diamide, monoamide carboxylic acid and dicarboxylic acid by biotransformation has the characteristics of simple and convenient operation, high reaction efficiency, mild reaction conditions, high enantioselectivity, easy separation of products and high product purity, and is used for synthesizing novel pseudo nucleoside.

Description

Chiral pseudonucleoside compound and its preparation method and application

technical field

The invention relates to a chiral pseudonucleoside compound, a preparation method and application thereof.

Background technique

Pseudo-nucleoside compounds, also known as unnatural nucleoside compounds, are a class of compounds that are similar in chemical structure to natural nucleosides, so they can interfere with or be directly used in the metabolic process of nucleic acids in vivo, and block the synthesis of proteins and nucleic acids. Biosynthesis, therefore, these compounds occupy an important position in antiviral and antitumor drugs.

At present, the pseudonucleoside compounds mainly use natural sugar compounds as raw materials, and the pseudonucleoside compounds are synthesized through the formation of glycosidic bonds. Among them, the selective control of glycosidic bonds is more difficult. In the present invention, a cis or trans chiral pseudosugar ring skeleton is first constructed through a biocatalytic method, thereby avoiding the selective control of glycosidic bonds.

Biocatalysis is by far the most efficient, highly selective and environmentally friendly process, and the use of biocatalysis to synthesize some high value-added chemicals, especially chiral chemicals, has important application prospects and significance. Nitrile is an important intermediate in organic synthesis. The chemical transformation of nitrile requires harsh conditions and poor selectivity, while the biotransformation reaction of nitrile has the advantages of mild conditions and high selectivity. At present, it has been applied industrially to prepare the corresponding carboxylic acid and Amide derivatives, most famously, in 1985, the Japanese Nitto Company (now renamed as Mitsubishi Rayon Company) took the lead in realizing the industrialization of microbial synthesis of acrylamide. In 1998, the annual output of acrylamide exceeded 40,000 tons, making it one of the largest industrialized biotransformation pathways in the world. Rhodococcus rhodochrous J1 is also used by Swiss Lonza AG company to industrially produce B vitamins nicotinamide and niacin, and the annual output of nicotinamide has exceeded 3,000 tons. However, compared with the enzyme-catalyzed ester hydrolysis and ester bond formation reactions, the biocatalytic reactions of nitriles and amides are relatively few. The biotransformation reaction of nitriles and amides can not only obtain carboxylic acids, but also can obtain nitrogen-containing organic compounds amides or nitriles that cannot be obtained by ester hydrolysis through kinetic resolution and desymmetrization.

Contents of the invention

The object of the present invention is to provide a chiral pseudonucleoside compound and its preparation method and application.

The chiral pseudonucleoside compound provided by the present invention is a compound shown in formula I;

(Formula I)

In the formula I, n is an integer of 1-3, and m is 0 or 1;

*Represents chirality, R or S configuration;

X is -OH, Wherein, R ¹ is an alkyl or phosphoric acid group with a total of 1-5 carbon atoms, and R ⁴ is selected from -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and - At least one of HSO ₃ ;

Y is -OH, Wherein, R ⁴ is selected from at least one of -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and -HSO ₃ ;

R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH _{3 , -CH2CH2COOCH3 , -CH2CH2CONH2 , -CH2CH2CHO , or -CH2CH2CN .}

In the preparation shown in the above formula I provided by the present invention, m is 0 and Y is X is -OH, The method of the pseudonucleoside compound is method a or method b, wherein, method a uses the compound shown in formula II as the starting reactant, comprising the steps of:

1) react the compound described in formula II' with SOCl ₂ in N, N-dimethylformamide, and the compound shown in formula IIa1 is obtained after the reaction is completed;

(Formula II')

In the formula II', n is an integer of 1-3; * represents chirality, and is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO _{or -CH2CH2CN} . ; R ₃ is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ ;

(Formula IIa1)

2) The compound shown in the formula IIa1 obtained in step 1) is refluxed with NaN _{and ZnBr} in a mixed solution composed of isopropanol and water, and the compound shown in the formula IIb1 is obtained after the reaction is completed;

(Formula IIb1)

3) Refluxing the compound shown in formula IIb1 obtained in step 2) with LiAlH _in tetrahydrofuran for reflux reaction, and the compound shown in formula IIc1 is obtained after the reaction is completed;

(Formula IIc1)

4) The compound shown in formula IIc1 obtained in step 3) is reacted with acetic anhydride, pyridine and 4-dimethylamino-pyridine in dichloromethane. After the reaction is completed, m is 0 and Y is X is -OH, A pseudonucleoside compound, that is, a compound shown in formula IId1;

(Formula IId1)

In the formula IIa1, formula IIb1, formula IIc1 and formula IId1, n is an integer of 1-3; * represents chirality, which is R or S

In the step 1) of the method, the compound of the formula II', SOCl _and N,N-dimethylformamide are used in an amount ratio of 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferably 10mmol : 1mL: 4mL; in the reaction step, the temperature is -10°C-50°C, preferably 0°C, and the time is 0.5-24 hours, preferably 1 hour; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 2), the ratio of the compound represented by the formula IIa1, NaN ₃ , ZnBr ₂ to the mixed solution composed of isopropanol and water is 0.1-10.0 mmol: 0.2-20 mmol: 0.1-10 mmol: 1 -10mL, preferably 1mmol: 2mmol: 0.5mmol: 4.5mL; in the mixed solution composed of isopropanol and water, the volume ratio of isopropanol and water is 1-10: 0.1-100, preferably 1: 2; In the reflux reaction step, the time is 1-24 hours, preferably 16 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 3), the compound shown in the formula IIb1, _LiAlH and tetrahydrofuran are used in an amount ratio of 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferably 1mmol: 10mmol: 10mL; in the reflux reaction step, the time It is 6-48 hours, preferably 12 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 4), the dosage ratio of the compound represented by the formula IIc1, acetic anhydride, pyridine, 4-dimethylaminopyridine, and dichloromethane is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL , preferably 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; the reaction temperature is 0-50°C, preferably 25°C, and the time is 6-48 hours, preferably 12 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol ;

Method b takes the compound shown in formula III' as starting reactant, comprises the steps:

Described method b comprises the steps:

1) react the compound described in the formula III' with SOCl in DMF, _and the compound shown in the formula IIIa1 is obtained after the reaction is completed;

(Formula III')

In the formula III', n is an integer of 1-3; * represents chirality, which is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN; R ³ is -NH ₂ , -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH=CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ , R ³ 'is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ ;

(Formula IIIa1)

2) The compound shown in the formula IIIa1 obtained in step 1) is refluxed with NaN _{and ZnBr} in a mixed solution composed of isopropanol and water, and the compound shown in the formula IIIb1 is obtained after the reaction is completed;

(Formula IIIb1)

3) The compound shown in the formula IIIb1 obtained in step 2) is refluxed with _LiAlH in tetrahydrofuran, and the compound shown in the formula IIIc1 is obtained after the reaction is completed;

(Formula IIIc1)

4) The compound shown in formula IIIc1 obtained in step 3) is reacted with acetic anhydride, pyridine and 4-dimethylamino-pyridine in dichloromethane, and after the reaction is completed, m is 0 and Y is X is -OH, The pseudonucleoside compound, that is, the compound shown in formula IIId1,

(Formula IIId1)

In the formula IIIa1, formula IIIb1, formula IIIc1 and formula IIId1, n is an integer of 1-3; * represents chirality, which is R or

In step 1) of the above method, the compound of the formula III', SOCl and DMF are used in an amount ratio of 1.0-10.0mmol: _0.1-2mL : 1-100mL, preferably 10mmol: 1mL: 4mL; in the reaction step , the temperature is -10°C-50°C, preferably 0°C, and the time is 0.5-24 hours, preferably 1 hour; according to the above optimized conditions, the reaction scale ranges from 0.5mmol-100mmol;

In the step 2), the dosage ratio of the compound represented by the formula IIIa1, NaN ₃ , ZnBr ₂ to the mixed solution composed of isopropanol and water is 0.1-10.0 mmol: 0.2-20 mmol: 0.1-10 mmol: 1 -10mL, preferably 1mmol: 2mmol: 0.5mmol: 4.5mL; in the mixed solution composed of isopropanol and water, the volume ratio of isopropanol and water is 1-10: 0.1-100, preferably 1: 2; In the reflux reaction step, the time is 1-24 hours, preferably 16 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 3), the compound shown in the formula IIIb1, _LiAlH and tetrahydrofuran are used in an amount ratio of 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferably 1mmol: 10mmol: 10mL; in the reflux reaction step, the time It is 6-48 hours, preferably 12 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 4), the dosage ratio of the compound represented by the formula IIIc1, acetic anhydride, pyridine, 4-dimethylaminopyridine and dichloromethane is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL , preferably 1mmol: 10mmol: 1mmol: 0.1mmol: 10mL; the reaction temperature is 0-50°C, preferably 25°C, and the time is 6-48 hours, preferably 12 hours. According to the above optimization conditions, the reaction scale range is 0.5mmol-100mmol;

In the preparation shown in the above formula I provided by the present invention, m is 1, Y is -OH or X is -OH, The method for pseudonucleoside compounds, comprising the steps of:

1) Refluxing the compound described in formula II' with LiAlH ₄ in tetrahydrofuran, the reaction is completed to obtain the compound shown in formula IIa2;

(Formula II')

In the formula II' and formula IIa2, n is an integer of 1-3; * represents chirality, and is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , - CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN; R ³ is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH=CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ ;

(Formula IIa2)

2) react the compound shown in formula IIa2 obtained in the step 1) with R ⁴ PhCH ₂ CH ₂ CHO and NaBCNH ₃ in dichloromethane or methanol, and the compound shown in formula IIb2 is obtained after the reaction is completed; the R ⁴ PhCH ₂ In CH ₂ CHO, R ⁴ is selected from at least one of -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and -HSO ₃ ;

(Formula IIb2)

In the formula IIb2, n is an integer of 1-3; * represents chirality, and is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , - CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or _{-CH2CH2CN .} ; R ⁴ is selected from at least one of -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and -HSO ₃ ; Ph is phenyl;

3) The compound shown in the formula IIb2 obtained in the step 2) is refluxed with an aqueous solution of hydrochloric acid and formaldehyde, and the compound shown in the formula IIc2 is obtained after the reaction is completed, that is, in the formula I, m is 1, Y is -OH, X for compound of;

(Formula IIc2)

In the formula IIc2, n is an integer of 1-3; * represents chirality, and is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , - CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or _{-CH2CH2CN .} ; R ⁴ is selected from at least one of -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and -HSO ₃ ;

4) The compound shown in formula IIc2 obtained in step 3) is reacted with acetic anhydride, pyridine and 4-dimethylamino-pyridine in dichloromethane. After the reaction is completed, m is 1 and Y is X is -OH, The pseudonucleoside compound, that is, the compound shown in formula IId2,

(Formula IId2)

In the formula IId2, n is an integer of 1-3; * represents chirality, and is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , - CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or _{-CH2CH2CN .} ; R ⁴ is selected from at least one of -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and -HSO ₃ ; X is -OH, R ¹ is an alkyl group or a phosphoric acid group with a total of 1-5 carbon atoms.

In the step 1) of the above-mentioned method, the dosage ratio of the compound described in the formula II', LiAlH4 and tetrahydrofuran _is 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferably 1mmol: 10mmol: 10mL; in the reflux reaction step , the time is 6-48 hours, preferably 24 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 2), the dosage ratio of the compound represented by the formula IIa2, R ⁴ PhCH ₂ CH ₂ CHO, NaBCNH ₃ and dichloromethane or methanol is 0.1-10mmol: 0.1-10mmol: 1-100mmol: 1-100mL , preferably 1mmol: 0.9mmol: 2mmol: 6mL; in the reaction step, the temperature is 0-50°C, preferably 25°C, and the time is 12-48 hours, preferably 24 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol- 100mmol;

In the step 3), the dosage ratio of the compound represented by the formula IIb2, the aqueous solution of formaldehyde, hydrochloric acid and chloroform is 0.1-10mmol: 0.1-20mL: 1-40mL: 0-40mL, preferably 1mmol: 2.4mL: 4.8mL : 4.8mL; in the reflux reaction step, the time is 1-48 hours, preferably 24 hours; the mass percent concentration of the hydrochloric acid is 10-35%, preferably 35%; the mass percent concentration of the aqueous solution of formaldehyde 10-35%, preferably 35%; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 4), the compound shown in formula IIc2, acetic anhydride, pyridine, 4-dimethylaminopyridine, and methylene chloride are used in an amount ratio of 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferably 1mmol : 10mmol: 1mmol: 0.1mmol: 10mL; the reaction temperature is 0-50°C, preferably 25°C, and the time is 6-48 hours, preferably 12 hours. According to the above optimization conditions, the reaction scale range is 0.5mmol-100mmol;

In the preparation of the above formula I provided by the present invention, m=1, X is -OH, Y is -OH or The method for pseudonucleoside compounds, comprising the steps of:

1) the compound described in formula II' is reacted with dimethoxycyanuric chloride, nitrogen methylmorpholine and tetrahydroisoquinoline in methylene chloride, and the compound shown in formula IIa3 is obtained after the reaction is completed;

(Formula II')

In the formula II' and formula IIa3, n is an integer of 1-3; * stands for chirality, which is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , - CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN; R ³ is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH=CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ ; R ⁴ is selected from at least one of -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and -HSO ₃ ;

(Formula IIa3)

₂ ) reacting the compound shown in the formula IIa3 obtained in step 1) with SOCl in DMF, the reaction is completed to obtain the compound shown in the formula IIb3;

(Formula IIb3)

3) The compound shown in formula IIb3 obtained in step 2) and the mixed solution composed of methanol and dichloromethane are passed into dry HCl gas for reaction, and the compound shown in formula IIc3 is obtained after the reaction is completed;

(Formula IIc3)

4) The compound shown in the formula IIc3 obtained in step 3) is refluxed with _LiAlH in tetrahydrofuran, and the compound shown in the formula IId3 is obtained after the reaction is completed, that is, m=1 and X in the formula I are Y is a compound of -OH;

(Formula IId3)

5) The compound shown in formula IId3 obtained in step 3) is reacted with acetic anhydride, pyridine and 4-dimethylamino-pyridine in dichloromethane, and the reaction is completed to obtain m=1 in the formula I, and X is- Oh, Y is The pseudonucleoside compound, that is, the compound shown in formula IIe3,

(Formula IIe3)

In the formula IIb3-Formula IIe3, n is an integer of 1-3; * represents chirality, which is R or S configuration; R ² is -H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , -Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN; X is -OH, R ¹ is an alkyl or phosphoric acid group with a total of 1-5 carbon atoms, and R ⁴ is selected from -H, -F, -Cl, -Br, -I, -CH ₃ , -OCH ₃ , -NO ₂ and -HSO At least one of ₃ .

In step 1) of the above-mentioned method, the dosage ratio of the compound described in the formula II', CDMT (dimethoxycyanuric chloride), NMM (nitrogen methylmorpholine), tetrahydroisoquinoline and dichloromethane 0.1-10mmol: 0.1-20mmol: 0.1-20mmol: 0.1-10mmol: 1-40mL, preferably 5mmol: 1.2mmol: 3mmol: 1.1mmol: 10mL; in the reaction step, the temperature is 0-50°C, preferably 25°C , the time is 6-48 hours, preferably 12 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 2), the compound shown in the formula _IIa3 , SOCl and DMF are used in an amount ratio of 1.0-10.0mmol: 0.1-2mL: 1-100mL, preferably 4mmol: 1mL: 4mL; in the reaction step, the temperature It is -10°C-50°C, preferably 0°C, and the time is 0.5-24 hours, preferably 1 hour; according to the above optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 3), the compound shown in the formula IIb3 and the mixed solution composed of methanol and dichloromethane are used in an amount ratio of 0.1-1mmol: 1-15mL, preferably 0.5mmol: 15mL; the introduction of the dry HCl gas The volume is 22.4mL-22.4L, preferably 1L; in the reaction step, the temperature is -10°C-50°C, preferably 0°C, and the time is 0.5-24 hours, preferably 1 hour; the reaction is composed of methanol and dichloromethane In the mixed solution, the volume ratio of methanol and dichloromethane is 2:1; in the reaction step, the temperature is -20-0°C, preferably -20°C, and the time is 0.5-12 hours, preferably 1 hour; according to the above The reaction scale range of optimized conditions is 0.5mmol-100mmol;

In the step 4), the compound shown in the formula IIc3, _LiAlH4 and tetrahydrofuran are used in an amount ratio of 0.1-10mmol: 0.5-50mmol: 1mL-20mL, preferably 0.1mmol: 10mmol: 4mL; in the reflux reaction step, The time is 6-48 hours, preferably 24 hours; according to the above-mentioned optimized conditions, the reaction scale range is 0.5mmol-100mmol;

In the step 5), the compound represented by formula IId3, acetic anhydride, pyridine, 4-dimethylaminopyridine, and methylene chloride are used in an amount ratio of 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL, preferably 1mmol : 10mmol: 1mmol: 0.1mmol: 10mL; the reaction temperature is 0-50°C, preferably 25°C, and the time is 6-48 hours, preferably 12 hours. According to the above optimization conditions, the reaction scale range is 0.5mmol-100mmol;

The present invention also provides the compound shown in the starting material compound formula II and formula III for preparing the compound shown in the above formula I, wherein the compound shown in formula II is as follows,

(Formula II)

In the formula II, n is an integer of 1-3; * represents chirality, and is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH 2 CH=CH 2 , -CH ₂ CH=CH ₂ , - CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or _{-CH2CH2CN .} ; R ³ is -NH ₂ , -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ .

The compound shown in the formula II is the compound shown in the formula II',

(Formula II')

In the formula II', n is an integer of 1-3; * represents chirality, which is R or S configuration; R ² is -H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , -Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN; R ³ is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH=CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ .

The method for preparing the compound shown in the above formula II provided by the present invention is obtained by desymmetrizing catalytic hydrolysis and esterification, and the reaction equation is as follows:

(Formula IV)

The preparation method is the following method a or method b, wherein, the method a comprises the following steps: after carrying out a catalytic hydrolysis reaction with a compound shown in formula IV with a Rhodococcus erythropolis AJ270 catalyst system, and then reacting with alkali and benzyl bromide Or carry out esterification reaction to methoxybenzyl bromide in a solvent, after the reaction is completed, the compound shown in the formula II is obtained,

(Formula IV)

In the formula IV, R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH 2 CH═CH ₂ , _{-CH 2 C 6} H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN, n is an integer of 1-3;

Said method b comprises the following steps: after performing a catalytic hydrolysis reaction with a compound shown in Formula IV with a Rhodococcus erythropolis AJ270 catalyst system, and then carrying out an esterification reaction with diazomethane ether solution, the reaction is completed to obtain the formula II Compound shown,

(Formula IV)

In the formula IV, R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH 2 CH═CH ₂ , _{-CH 2 C 6} H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN, n is an integer of 1-3.

In the method a and method b, the Rhodococcus catalytic system is composed of Rhodococcus and a buffer solution with a pH value of 6.0-8.0, specifically connecting the Rhodococcus to the buffer solution with a pH value of 6.0-8.0 Activated at 30°C for 30 minutes; the buffer solution is Na ₂ HPO ₄ -citric acid buffer solution, K ₂ HPO ₄ -KH ₂ PO ₄ buffer solution, Tris buffer solution, Hanks' buffer solution or PBS buffer solution; The dosage ratio of Rhodococcus to the compound shown in formula IV is 2g: 1mmol-2g: 1mol; in the Rhodococcus catalytic system, the dosage ratio of Rhodococcus to the buffer solution is 2g: 50mL-2g: 1L; In the catalytic hydrolysis reaction step, the temperature is 20-37°C, preferably 30°C, and the time is 0.1-120 hours. Different substrates and dosages are preferably different times, so that the enantioselectivity of the reaction product is above 95%;

In the method a, the alkali is potassium carbonate, sodium carbonate, potassium hydroxide, sodium hydroxide or cesium carbonate, preferably potassium carbonate; the solvent is selected from acetone, N, N-dimethylformamide, dimethyl At least one of base sulfoxide and tetrahydrofuran, preferably N,N-dimethylformamide; the amount ratio of the base, the solvent, benzyl bromide or p-methoxybenzyl bromide to the compound shown in the formula IV 1.38g: 2mL: 1mL: 1mmol; in the esterification step, the temperature is 10-30°C, preferably 25°C, and the time is 1-48 hours, preferably 24 hours;

In the method b, the concentration of the ether solution of diazomethane is 0.1-1mol/L, preferably 0.5mol/L; in the esterification step, the temperature is -20-10°C, preferably 0°C, and the time 2-24 hours, preferably 12 hours.

The present invention also provides the compound shown in formula III,

(Formula III)

In the formula III, n is an integer of 1-3; * represents chirality, which is R or S configuration; R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH 2 CH=CH 2 , -CH ₂ CH=CH ₂ , - CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or _{-CH2CH2CN .} ; R ³ is -NH ₂ , -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ , R ³ ' is -NH ₂ , -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ .

The compound described in the formula III is a compound shown in the formula III',

(Formula III')

In the formula III', n is an integer of 1-3; * represents chirality, which is R or S configuration; R ² is -H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH=CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , -Cbz, -CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN; R ³ is -NH ₂ , -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH=CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ , R ³ ′ is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ .

The method for preparing the compound shown in the above formula III provided by the present invention is obtained through dynamic resolution and esterification, and its reaction equation is as follows:

The preparation method is the following method a or method b, wherein said method a comprises the following steps: after catalytic hydrolysis reaction with the compound shown in formula V with Rhodococcus (Rhodococcus erythropolis AJ270) catalytic system, then react with alkali and methyl iodide , benzyl bromide or p-methoxybenzyl bromide carry out esterification reaction in a solvent, the reaction is completed to obtain the compound shown in the formula III,

(Formula V)

In the formula V, R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH═CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, - CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN, n is an integer of 1-3;

Said method b comprises the following steps: after performing a catalytic hydrolysis reaction with a compound shown in formula V with a Rhodococcus erythropolis AJ270 catalyst system, and then performing an esterification reaction with diazomethane ether solution, the reaction is completed to obtain said formula III Compound shown,

(Formula V)

In the formula V, R ² is H, -CH ₃ , -CH ₂ CH ₃ , -CH ₂ CH═CH ₂ , -CH ₂ C ₆ H ₅ , -CH ₂ C ₆ H ₅ OCH ₃ , Cbz, - CH ₂ CH ₂ COCH ₃ , -CH ₂ CH ₂ COOCH ₃ , -CH ₂ CH ₂ CONH ₂ , -CH ₂ CH ₂ CHO or -CH ₂ CH ₂ CN, n is an integer of 1-3.

In the method a and method b, the Rhodococcus catalytic system is composed of Rhodococcus and a buffer solution with a pH value of 6.0-8.0, specifically connecting the Rhodococcus to the buffer solution with a pH value of 6.0-8.0 Activated at 30°C for 30 minutes; the buffer solution is Na ₂ HPO ₄ -citric acid buffer solution, K ₂ HPO ₄ -KH ₂ PO ₄ buffer solution, Tris buffer solution, Hanks' buffer solution or PBS buffer solution; The dosage ratio of Rhodococcus to the compound shown in formula IV is 2g: 1mmol-2g: 1mol; in the Rhodococcus catalytic system, the dosage ratio of Rhodococcus to the buffer solution is 2g: 50mL-2g: 1L; In the catalytic hydrolysis reaction step, the temperature is 20-37°C, preferably 30°C, and the time is 0.1-120 hours. Different substrates and dosages are preferably different times, so that the enantioselectivity of the reaction product is above 95%;

In the method a, the alkali is potassium carbonate, sodium carbonate, potassium hydroxide, sodium hydroxide or cesium carbonate, preferably potassium carbonate; the solvent is selected from acetone, N, N-dimethylformamide, dimethyl At least one of base sulfoxide and tetrahydrofuran, preferably N,N-dimethylformamide; the amount ratio of the base, the solvent, benzyl bromide or p-methoxybenzyl bromide to the compound shown in the formula IV 1.38g: 2mL: 1mL: 1mmol; in the esterification step, the temperature is 10-30°C, preferably 25°C, and the time is 1-48 hours, preferably 24 hours;

In the method b, the diazomethane diethyl ether solution has a concentration of 0.1-1mol/L, preferably 0.5mol/L; in the esterification step, the temperature is -20-10°C, preferably -15°C, The time is 2-24 hours, preferably 12 hours.

The biocatalytic substrate preparation methods in the above two microbial catalysis-esterification processes can be divided into the following three categories according to the literature sources.

The first category: the preparation of cis and formula V trans-N-substituted 2,5-disubstituted tetrahydropyrrole diamide compounds shown in the above formula IV, wherein n=1. These compounds are prepared by chemical hydrolysis of 2,5-disubstituted tetrahydropyrrole dinitrile compounds. Nitriles were prepared according to the methods given in Takahashi, K.; Saitoh, H.; Ogura, K.; Iida, H. Heterocycles 1986, 24, 10, 2905.

The second category: the preparation of cis- and trans-2,6-disubstituted hexahydropiperidine diamide compounds shown in the above formula IV, wherein n=2. Prepared according to the following literature methods: Takahashi K.; Mikajiri T.; Kurita H.; Ogura K.; Iida H., J.Org.Chem.1985, 50, 4372-4375.

The third category: the preparation of cis-form and formula V-trans-2,7-disubstituted nitrogen seven-membered cyclic diamide compounds shown in the above formula IV, wherein n=3. According to literature methods: Ogura K.; Shimamura Y.; Fujita M., J. Org. Chem. 1991, 56, 2920-2922.

The Rhodococcus erythropolis AJ270 sample used in the above two microbial catalyzed-esterification processes was originally isolated from an acetonitrile agar medium containing 25mM using an Anderson bioparticle sampler, and the sample source was originally collected in the River Tyne, UK In dry soil near an abandoned industrial factory. The chemical taxonomic study of mycolic acid and diaminopimelic acid on the cell wall of AJ270 bacteria confirmed that it belonged to Rhodococcus species. Until 2005, through the study of its 16SrRNA gene sequence, it was confirmed that Rhodococcus AJ 270 belonged to Rhodococcus erythropolis strain. Specifically refer to the following two 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 van Leeuwenhoek 2005, 87, 221-232.

Rhodococcus AJ 270 is a soil-derived microorganism and has been shown to be a highly active whole-cell catalyst of the nitrile hydratase/amidohydrolase system. Previous studies have shown that, compared with other strains, Rhodococcus erythropolis AJ270 has a broad spectrum of substrates and can efficiently catalyze the hydrolysis of aliphatic nitriles, aromatic nitriles and aromatic heterocyclic nitriles. (Wang M.-X. Enantioselective biotransformations of nitrogen in organic synthesis. Top. Catal. 2005, 35, 117-130)

In addition, the antitumor drugs with the pseudonucleoside compounds provided by the present invention as active ingredients belong to the protection scope of the present invention. In actual operation, in order to achieve better drug efficacy, various adjuvants can be added to the above pseudonucleoside compounds as required. In addition, the application of the pseudonucleoside compounds provided by the present invention in the preparation of antitumor drugs also belongs to the protection scope of the present invention. The tumor is liver cancer or lung cancer. The above-mentioned antitumor drugs can be drugs against Bel-7402 liver cancer cells or A-549 lung cancer cells.

The raw materials for preparing pseudonucleoside compounds provided by the invention are obtained by catalyzing the hydrolysis of nitrogen heterocyclic diamide compounds by Rhodococcus erythropolis AJ270 microbial system. The amount of rhodococcus thalli used can be adjusted according to the amount of substrate. The reaction solvent is a common buffer solution with pH=6.0-8.0, the temperature is 20-37° C., and the reaction time is 0.1-120 hours. The rhodococcus microbial catalytic system has the characteristics of fermentable culture and convenient preservation. Using this biotransformation method to prepare chiral nitrogen heterocyclic diamides, monoamide carboxylic acids, and dicarboxylic acids has the advantages of simple operation, high reaction efficiency, mild reaction conditions, high enantioselectivity, easy separation of products, and high product purity. characteristics, and used for the synthesis of new pseudonucleosides.

Detailed ways

The present invention will be further described below in conjunction with specific examples, but the present invention is not limited to the following examples. The methods are conventional methods unless otherwise specified. The reactants can be obtained from public commercial sources unless otherwise specified. The Chinese name of Cbz in the present invention is benzyloxycarbonyl.

Embodiment 1: the synthesis of pseudonucleoside compound 1 '

1) Dissolve 10 mmol of the compound X' (Cbz, Chinese name benzyloxycarbonyl) of the formula II' prepared in Example 15 in 4 ml of dry DMF, add 1 ml of thionyl chloride SOCl ₂ , and stir at 0°C for 1 hour , According to TLC monitoring, the reaction starting material had disappeared. The reaction solution was extracted with ethyl acetate and subjected to ethyl acetate silica gel column chromatography to obtain the monocyanomonocarboxylate of the compound shown in the formula IIa1 (that is, the compound shown in 1-1 in the reaction formula), an oily liquid, and the yield was 90 %.

2) Take out 2mmol step 1) Gained single cyano monocarboxylate is put into 25 milliliters of eggplant-shaped bottles, add 4mmol sodium azide NaN ₃ , 1mmol zinc bromide ZnBr ₂ and by 6 milliliters of water and 3 milliliters of isopropyl The mixed solution composed of alcohol was refluxed for 16 hours. After cooling down, adjust the pH to less than 7 with 1N HCl, and extract with ethyl acetate. Then use silica gel column chromatography, eluent is ethyl acetate: methanol=20: 1 (volume ratio, 1% acetic acid as additive), receive colorless oily matter, be the compound shown in formula IIb1, (also be reaction The compound shown in 1-2 in the formula, which is the precursor of compound 1), has a yield of 58%.

3) Take 1 mmol of the compound shown in 1-2 obtained in step 2), add 10 mmol of lithium aluminum hydrogen and 10 mL of tetrahydrofuran to reflux for 12 hours, and purify with a cation exchange resin to obtain the compound shown in formula IIc1 (that is, compound 1 in the reaction formula), which is white Solid, 83% yield.

Compound 1: solid, mp 185°C (D ₂ O/300MHz) δ3.86-3.75(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 ₂ O) δ163.0, 67.6, 63.6, 62.5, 39.5, 28.6, 26.8. (KBr) 3432, 3259, 1634 , 1023cm ^-1 ; MS (ESI) m/z 184.1 (M ⁺ +1, 15%), 206.1 (M ⁺ +23, 100%). Anal.Calcd.for: [M+H] C ₇ H ₁₄ N ₅ O, 184.1198, Found: [M+H] 184.1193.

It can be seen from the above that the structure of this compound is correct, which is compound 1 in the reaction formula.

4) Take 1 mmol of compound 1 obtained in step 3), dissolve it in 10 mL of dichloromethane, add 10 mmol of acetic anhydride, 1 mmol of pyridine, and 0.1 mmol of 4-dimethylaminopyridine, and stir at 25°C for 12 hours to obtain the compound provided by the invention Pseudonucleoside compound 1' shown in formula I, wherein, n is 1, m is 0, and Y is R ² is -CH ₃ , and X is -OCOCH ₃ .

Compound 1': solid, mp 105°C (D ₂ O/300MHz) δ3.91-3.70(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 ₂ O) δ162.5, 65.6, 62.6, 61.5, 39.2, 28.4, 27.2, 20.7.(KBr) 3432, 3259, 1634, 1023cm ^-1 ; Anal.Calcd.for: [M+H]C ₉ H ₁₅ N ₅ O ₂ , 226.1304, Found: [M+H] 226.1300.

As can be seen from the above, the compound has a correct structure and is a compound shown in formula I, that is, compound 1' in the reaction formula.

Embodiment 2: the synthesis of pseudonucleoside compound 2'

1) Dissolve 10 mmol of the compound X'' (Cbz, Chinese name benzyloxycarbonyl) of the formula III' prepared in Example 8 in 4 ml of dry DMF, add 1 ml of thionyl chloride SOCl ₂ , stir at 0°C for 1 Hours, the reaction starting material had disappeared according to TLC monitoring. The reaction solution was extracted with ethyl acetate and subjected to ethyl acetate silica gel column chromatography to obtain the compound monocyanomonocarboxylate (ie compound 2-1 in the reaction formula) represented by formula IIIa1 as an oily liquid with a yield of 90%.

2) Take out 2mmol step 1) Gained single cyano monocarboxylate is put into 25 milliliters of eggplant-shaped bottles, add 4mmol sodium azide NaN ₃ , 1mmol zinc bromide ZnBr ₂ and by 6 milliliters of water and 3 milliliters of isopropyl The mixed solution composed of alcohol was refluxed for 16 hours. After cooling down, the pH was adjusted to be less than 7 with 1N HCl, and extracted with ethyl acetate. Then use silica gel column chromatography, eluent is ethyl acetate: methyl alcohol=20: 1 (1% acetic acid is used as additive), receive white solid thing, be the compound shown in formula IIIb1 (that is, 2-2 in the reaction formula Shown compound, this compound is the precursor of compound 2), and the yield is 90%.

3) Take 1 mmol of the compound shown in 2-2 obtained in step 2), add 10 mmol of lithium aluminum hydride, reflux 10 mL of tetrahydrofuran for 12 hours, and purify with cation exchange resin to obtain the compound shown in formula IIIc1 (that is, compound 2 in the reaction formula), which is white Solid, 83% yield.

Compound 2: solid, mp 194°C (D ₂ O/300MHz) δ4.91-4.88(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 ₂ O) δ158, 67.0, 60.7, 59.3, 35.7, 27.4, 25.5.(KBr) 3432, 3259, 1634, 1023cm ^-1 ; MS (ESI) m/z 184.1 (M ⁺⁺ ₁ , 15%), 206.1 ₍ M ⁺⁺ 23, 100%).Anal.Calcd.for: [M+H] _C7H14N5 O, 184.1198, Found: [M+H] 184.1193.

It can be seen from the above that the structure of this compound is correct, which is compound 2 in the reaction formula.

4) Take 21mmol of the compound obtained in step 3), dissolve it in 10mL of dichloromethane, add 10mmol of acetic anhydride, 1mmol of pyridine, and 0.1mmol of 4-dimethylaminopyridine, and stir at 25°C for 12 hours to obtain the formula I provided by the present invention The shown pseudonucleoside compound 2', wherein, n is 1, m is 0, and Y is R ² is -CH ₃ , and X is -OCOCH ₃ .

Compound 2': solid, mp 154°C (D ₂ O/300MHz) δ4.77-4.50(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 ₂ O) δ155, 67.5, 60.2, 58.3, 33.7, 27.3, 24.5, 20.7.(KBr) 3432, 3259, 1634, 1023cm ^-1 ; Anal.Calcd.for: [M+H]C ₉ H ₁₅ N ₅ O ₂ , 226.1304, Found: [M+H] 226.1307.

It can be known from the above that the compound has a correct structure and is a compound shown in formula I (that is, compound 2 in the reaction formula).

Embodiment 3: the synthesis of pseudonucleoside compounds 3' shown in formula I

1) Dissolve 5 mmol of the compound X' of formula II' prepared in Example 15 in 50 ml of tetrahydrofuran, add 1.9 g (50 mmol) of lithium aluminum hydrogen, and reflux for 24 hours, then add 2N aqueous sodium hydroxide solution to quench the reaction. Diatomaceous earth suction filtration, cation exchange resin purification, the primary amine compound shown in formula IIa2 (specifically the compound shown in 3-1 in the above reaction equation) was obtained, with a yield of 79%.

2) Mix 0.625 mmol of the primary amine compound shown in formula IIa2 obtained in step 1) with 0.6 mmol of phenylacetaldehyde in 4 ml of dichloromethane, add 1.2 mmol of sodium cyanoborohydride, and react at 25°C for 24 hours, then use 1N HCl The product was extracted from the organic phase, then the pH was adjusted to greater than 7, and the product was extracted from water with ethyl acetate. Further, cation exchange resin chromatography was used to obtain the compound shown in formula IIb2 (specifically, the compound shown in 3-2 in the above reaction equation), with a yield of 54%.

3) Take 0.5 mmol of the compound shown in the obtained formula IIb2 in step 2), add 1.2 milliliters of formaldehyde solution with a mass percentage concentration of 35%, 2.4 milliliters of mass percentage concentration of 35% concentrated hydrochloric acid, and 2.4 milliliters of chloroform, and reflux for 24 hours to react After completion, the pH value of the adjustment system was 9, extracted with dichloromethane, dried over anhydrous sodium sulfate, ODS conventional column chromatography, mobile phase was methanol: water=1: 1, and the compound shown in formula IIc2 was obtained after the chromatography was completed (specifically The compound shown in 3 in the above reaction equation), the yield is 78%.

Compound 3: oil, ¹ HNMR (D ₂ O/300MHz) δ7.11-6.99 (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); ¹³ CNMR(75MHz/ CDCl ₃ ) δ134.9, 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 -1} ; MS (ESI) m/z 261.4 (M ⁺ +1, 100%), Anal. Calcd. for [M+H] C ₁₆ H ₂₅ N ₂ O, 261.1967, Found: [M+H] 261.1961.

It can be seen from the above that the structure of this compound is correct, which is compound 3 in the reaction formula.

4) Dissolve 1 mmol of the compound shown in formula IIc2 obtained in step 3) in 10 mL of dichloromethane, add 10 mmol of acetic anhydride, 1 mmol of pyridine, and 0.1 mmol of 4-dimethylaminopyridine, and stir and react at 25°C for 12 hours. After the reaction is completed, The pseudonucleoside compound 3' represented by formula I provided by the present invention.

In the pseudonucleoside compound 3' shown in the formula I, n is 1, R ² is -CH ₃ , m is 1, and Y is Wherein, R ⁴ is H, and X is -OCOCH ₃ .

Compound 3': oil, ¹ HNMR (D ₂ O/300MHz) δ7.21-6.95 (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); ¹³ CNMR(75MHz /CDCl ₃ ) δ134.5, 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 ^-1 ; MS (ESI) m/z 261.4 (M ⁺ +1, 100%), Anal. Calcd. for [M+H] C ₁₈ H ₂₆ N ₂ O ₂ , 303.2073, Found: [M+H] 303.2070.

It can be known from the above that the compound has a correct structure and is a compound shown in formula I.

Embodiment 4: the synthesis of pseudonucleoside compounds 4' shown in formula I

1) Dissolve 5 mmol of the compound X' shown in Formula II' prepared in Example 15 in 10 ml of dichloromethane, add 1.2 mmol of dimethoxycyanuric chloride (CDMT), 3 mmol of nitrogen methylmorpholine (NMO) After stirring at room temperature for 1 hour, 1.1 mmol tetrahydroisoquinoline (THIQ) was added, and stirring was continued overnight. Add 1N HCl aqueous solution to quench the reaction, dichloromethane extraction, anhydrous sodium sulfate drying, ethyl acetate column chromatography, receive the glassy compound, which is the primary amine compound shown in the formula Iia3 (specifically 4- in the above reaction equation) 1), the yield was 90%.

2) Dissolve 4 mmol of the compound shown in 4-1 obtained in step 1) in 4 ml of dry DMF, and add 1 ml of thionyl chloride. Stir at 0° C. for 1 hour, and the reaction starting material has disappeared according to TLC monitoring. The reaction solution was extracted with ethyl acetate and subjected to ethyl acetate silica gel column chromatography to obtain the compound represented by formula IIb3 (specifically compound 4-2 in the above reaction equation) as a white solid with a yield of 78%.

3) Dissolve 0.5 mmol of the compound shown in 4-2 obtained in step 2) in 5 ml of dichloromethane, add 10 ml of methanol, freeze to -20°C, and feed dry hydrochloric acid gas for 1 hour while stirring, and measure it to 1 L. And stand overnight at -20°C. The reaction liquid was dropped into 6N aqueous hydrochloric acid solution cooled at 0°C. Adjust the pH to 7 with sodium bicarbonate and extract with ethyl acetate. Dry over anhydrous sodium sulfate. Through silica gel column chromatography, the compound represented by formula IIc3 (specifically compound 4-3 in the above reaction equation) was obtained with a yield of 58%.

4) Dissolve 0.1 mmol of the compound shown in 4-3 obtained in step 3) in 4 ml of tetrahydrofuran, add 10 mmol of lithium aluminum hydride, reflux for 24 hours, treat the reaction with 2N sodium hydroxide, extract with ethyl acetate, and then pass ODS Conventional column chromatography, the mobile phase is methanol: water = 1: 1. The compound represented by formula IId4 (specifically compound 4 in the above reaction equation) was obtained with a yield of 81%.

Compound 4: oil, ¹ HNMR (D ₂ O/300MHz) δ7.11-6.99 (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); ¹³ CNMR(75MHz/ CDCl ₃ ) δ134.9, 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 -1} ; MS (ESI) m/z 261.4 (M ⁺ +1, 100%), Anal. Calcd. for [M+H] C ₁₆ H ₂₅ N ₂ O, 261.1967, Found: [M+H] 261.1961.

5) Take out 1 mmol of compound 4 obtained in step 4), dissolve it in 10 mL of dichloromethane, add 10 mmol of acetic anhydride, 1 mmol of pyridine, and 0.1 mmol of 4-dimethylaminopyridine, and stir at 25° C. for 12 hours. After the reaction is completed, formula I is obtained Pseudo-nucleosides shown 4'.

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); ¹³ CNMR(75MHz/CDCl ₃ )δ134.5, 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 ^-1 ; MS (ESI) m/z 261.4 (M ⁺⁺ 1, 100%), Anal. Calcd. for [M+H]C ₁₈ H ₂₆ N ₂ O ₂ , 303.2073, Found: [M+H] 303.2070.

It can be known from the above that the compound has a correct structure and is the compound 4' shown in formula I, wherein n is 1, R ² is -CH ₃ , m is 1, and Y is Wherein, R ⁴ is H, and X is -OCOCH ₃ .

Embodiment 5: Compound 5, 6 and 7 shown in the preparation formula III

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) wash the thalline into a 150 ml Erlenmeyer flat-bottomed flask with a threaded opening, disperse and shake well, and put it into a shaker for activation at 30°C for 30 minutes. Millimoles (247 mg) of the substrate amide represented by formula V1 were placed in a shaker at 30° C. and 200 rpm for catalytic hydrolysis. The whole reaction was monitored by high performance liquid phase, and the reaction was stopped after 39 hours of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove bacteria, and the filter residue was washed three times with 20 ml of water successively.

2) The filtrate obtained in step 1) was freeze-dried (-50 to -60°C), soaked the residue with 2 ml of methanol, and then dripped the ether solution of diazomethane to carry out the methyl esterification reaction at -15°C for 12 hours , add a small amount of water after the end, then use ethyl acetate to extract, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, the eluent is ethyl acetate-ethyl acetate:methanol=20:1 , to obtain the diamide compound shown in the formula III provided by the present invention (specifically compound 5 in the above reaction formula) (yield: 49%, ee>99.5%, Dicel-ADH), monoamide carboxylate methyl ester (specifically 6) in the above reaction formula (yield: 39%, ee>99.5%, Dicel-ADH) and methyl dicarboxylate (specifically compound 7 in the above reaction formula) (yield: 6%, ee>99.5 %, Dicel-ODH).

5: solid mp 230°C; [α] _D ²⁵ =-38.0 (c2.0, CH ₃ OH); ¹ HNMR (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); ¹³ CNMR (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 ₁₃ H ₁₇ N ₃ O ₂ : C, 63.14; H, 6.93; N, 16.99. Found: C, 62.94; H, 6.92; N, 17.05.

6: solid mp 140°C; [α] _D ²⁵ =+121.5 (c4.0, CHCl ₃ ); ¹ HNMR (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); ¹³ CNMR(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, 1658 cm ^-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 ₁₄ H ₁₈ N ₂ O 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 ₆ H ₆ ); ¹ HNMR (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); ¹³ CNMR (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, 1679 cm ^-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 ₁₄ H ₁₈ N ₂ O ₃ : C, 64.10; H, 6.92; N, 10.68. Found: C, 64.25; H , 6.87; N, 10.92.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula III.

Embodiment 6: Compound 8, 9 and 10 shown in the preparation formula III

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) wash the thalline into a 150 ml Erlenmeyer flat-bottomed flask with a threaded opening, disperse and shake well, and put it into a shaker for activation at 30°C for 30 minutes. Millimoles (197 mg) of the substrate amide of the formula V2 were placed in a shaker at 30° C. and 200 rpm for catalytic hydrolysis. The whole reaction was monitored by high performance liquid phase, and the reaction was stopped after 7 hours of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove bacteria, and the filter residue was washed three times with 20 ml of water successively.

2) Freeze-dry the filtrate obtained in step 1) (-50 to -60°C), infiltrate the residue with 2 ml of methanol, and then drop into diazomethane diethyl ether solution with a concentration of 0.5mol/L at -15°C The methyl esterification reaction was carried out at 100-200 mesh silica gel column chromatography after concentration, and a small amount of water was added after the end, then extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated and chromatographed on a 100-200 mesh silica gel column, the eluent was ethyl acetate-acetic acid Ethyl ester:methanol=20:1, obtain the diamide compound shown in the formula III provided by the present invention (specifically compound 8 in the above reaction formula) (yield: 45%, ee>99.5%, Dicel-ADH), single Methyl amide carboxylate (specifically compound 9 in the above reaction formula) (36% yield, ee>96.8%, Dicel-ADH) and methyl dicarboxylate (specifically compound 10 in the above reaction formula) ( Yield: 10%, ee 93.3%, Dicel-OJH).

8: mp 222°C; [α] _D ²⁵ =-54.7 (c1.5, CH ₃ OH); ¹ HNMR (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); ¹³ CNMR (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, 1639 cm ⁻¹ ; MS (ESI) m/z 198.1 (M ⁺ +1, 100%), 220.2 (M ⁺ +23, 17.1). Anal. Calcd. for C ₉ H ₁₅ N ₃ O ₂ : C, 54.81; H, 7.67; N, 21.30. Found: C, 54.75; H, 7.71; N, 21.36.

9: mp 104°C; [α] _D ²⁵ =+78.0 (c1.0, CHCl ₃ ); ¹ HNMR (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); ¹³ CNMR(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 ₁₀ H ₁₆ N ₂ O 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 ); ¹ HNMR (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); ¹³ CNMR(75MHz/CDCl ₃ ) δ174.7, 135.3, 117.7, 63.6, 53.5, 51.6, 28.4; IR (KBr) 1738 cm ^-1 ; MS (EI) m/z 227 (M ⁺ , 3%), 169 (10), 168 (100%) , 108(11); Anal. Calcd. for C ₁₁ H ₁₇ NO ₄ : 227.1158 (M). Found: 227.1161.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula III.

Embodiment 7: Compound 5 and 11 shown in the preparation formula III

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) wash the thalli into a 150 ml Erlenmeyer flat-bottomed flask with a threaded opening, disperse and shake well, and then put it into a shaker for activation at 30°C for 30 minutes, then add 1 mmol (157 mg) of the formula V3 substrate at one time Amides were placed in a shaker at 30° C. and 200 rpm to carry out catalytic hydrolysis reaction. The entire reaction was monitored by TLC, and the reaction was stopped after 1 hour of reaction. The obtained reaction solution was filtered through a layer of diatomaceous earth to remove bacteria, and the filter residue was washed three times with 20 ml of water successively.

2) The filtrate obtained in step 1) was freeze-dried (-50 to -60°C), and the residue was mixed with 2 ml of N, N'-dimethyl diamide, 1 ml of benzyl bromide, and 1.38 g of potassium carbonate at room temperature for 24 hours. Add a small amount of water after the esterification reaction, then extract with ethyl acetate, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, the eluent is ethyl acetate-ethyl acetate:methanol=20 : 1, obtain the diamide compound (specifically compound 5 in the above-mentioned reaction formula) shown in the formula III provided by the invention (yield: 45%, ee＞99.5%, Dicel-ADH) and monoamide carboxylic acid benzyl ester ( Specifically, it is compound 11) in the above reaction formula (yield: 36%, ee>99.5%, Dicel-ADH).

11: solid; 104.0°C; IR(KBr)v 3396, 3172(CONH ₂ ), 1726, 1657, cm ^-1 ; ¹ H 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); ¹³ C 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 ₂₀ H ₂₂ N ₂ O ₃ : C, 70.99; H, 6.55; N, 8.28; Found: C, 70.66; H, 6.62; N, 8.43.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula III.

Embodiment 8, compound X shown in preparation formula III "

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) wash the thalline into a 150 ml Erlenmeyer flat-bottomed flask with a threaded opening, disperse and shake well, and put it into a shaker for activation at 30°C for 30 minutes. Millimoles (291 mg) of the substrate amide shown in formula V4 were placed in a shaker at 30° C. and 200 rpm for catalytic hydrolysis. The whole reaction was monitored by high-performance liquid phase, and the reaction was stopped after 39 hours of reaction to obtain a reaction solution containing the compound shown in the product formula X. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove the bacteria, and the filter residue was washed with 20 ml of water for three times. .

2) Freeze-dry the filtrate containing the compound represented by formula X obtained in step 1) (-50 to -60°C), infiltrate the residue with 2 ml of methanol, and then drop diazomethane in ether solution at -15°C The methyl esterification reaction was carried out at 100-200 mesh silica gel column chromatography after concentration, and a small amount of water was added after the end, then extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated and chromatographed on a 100-200 mesh silica gel column, the eluent was ethyl acetate-acetic acid Ethyl ester:methanol=20:1, to obtain methyl monoamide carboxylate (specifically X" in the above reaction formula), yield: 39%, ee>99.5%, Dicel-ADH.

X: solid mp 150°C; ¹ HNMR (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); ¹³ CNMR(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, 1044, 159 , 1352, 1229cm ^-1 ; MS (ESI) m/z 293.34 (M ⁺⁺ 1, 10%), 315.29 (M ⁺⁺ 23, 100%). Anal.Calcd.for: C ₁₄ H ₁₆ N ₂ O ₅ C, 57.53; H, 5.52; N, 9.58; Found: C, 57.75; H, 5.51, N, 9.58.

X": oil; ¹ HNMR (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); ¹³ CNMR(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, 28.3; IR (K, Br) 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 ₁₅ H ₁₈ N ₂ O ₅ C, 58.82; H, 5.92; N, 9.15 Found: C, 58.87; H, 5.99; N.9.10.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound X" shown in formula III.

Embodiment 9: Compounds 12 and 13 shown in the preparation formula II

Its reaction formula is as follows:

Specific implementation method:

1) Take 2 grams of wet weight of Rhodococcus erythropolis AJ270 bacteria (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ~250ml) into the Erlenmeyer flat-bottomed flask with threaded mouth, disperse and shake well, put it in a shaking table and activate it at 30°C for 30 minutes, then add 1 mmol to 200 mmol (157 mg to 30 grams) of the formula IV1 substrate amide, put it in a shaker at 30° C., and carry out catalytic hydrolysis reaction under the condition of 200 rpm. The entire reaction was monitored by TLC, and the reaction was stopped after 6 hours of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove bacteria, and the filter residue was washed three times with 20 ml of water successively.

2) Freeze-dry the filtrate obtained in step 1) (-50 to -60°C), purify it with an ion exchange resin to obtain a yellow solid, wash off the yellow impurities with methanol to obtain monoamide monocarboxylic acid 12 (yield: 92-96%, ee 96%, Dicel-ADH). The residue was then mixed with 2 ml of N,N'-dimethylformamide, 1 ml of benzyl bromide, and 1.38 g of potassium carbonate at room temperature at 25°C for 24 hours to carry out the esterification reaction. After the completion of the esterification reaction, a small amount of water was added, and then extracted with ethyl acetate. After drying over anhydrous sodium sulfate, concentrate and utilize 100-200 mesh silica gel column chromatography, the eluent is ethyl acetate, obtain the compound monoamide carboxylate benzyl ester shown in formula II provided by the present invention (specifically in the above reaction formula Compound 13) (yield: 96%, ee 96%, Dicel-ADH).

12: solid; mp 233-234°C IR(KBr)v 3308, 3153, 1696, 1642, 1447, 1403, 1330, 1308cm-1; ¹ HNMR(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); ¹³ NMR(75MHz/D2O) δ173.8, 171.4, 61.8, 60.3, 29.8, 29.2. MS (ESI) m/z 159.35 (M ⁺ +1, 7%), 181.46 (M ⁺ +23, 100%); Anal. Calcd. for C ₆ H ₁₀ N ₂ O 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°C; IR (KBr) v 3422, 3369 (CONH ₂ ), 1740, 1663 cm ^-1 ; ¹ H NMR (300 MHz, 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); ¹³ C 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 ₂₀ H ₂₂ N ₂ O ₃ : C, 70.99; H, 6.55; N, 8.28; Found: C, 70.65; H, 6.71; N, 8.53.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula II.

Embodiment 10: the compound 14 shown in preparation formula II

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) Wash the bacterium into a 150 ml Erlenmeyer flat-bottomed flask with screw top, disperse and shake well, put it in a shaker and activate it at 30°C for 30 minutes, then add 1 mg dissolved in 2.5 ml dimethyl sulfoxide at one time Mole (171 mg) of the formula IV2 substrate amide was put into a shaker at 30° C. and 200 rpm to carry out catalytic hydrolysis reaction. The entire reaction was monitored by TLC, and the reaction was stopped after 60 hours of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove bacteria, and the filter residue was washed with 20 ml of water for three times.

2) Freeze-dry the filtrate obtained in step 1) (-50 to -60°C), infiltrate the residue with 2 ml of methanol, and then drop into diazomethane diethyl ether solution with a concentration of 0.5mol/L at -15°C Carry out the methyl esterification reaction under the hood for 12 hours, add a small amount of water after the end, then use ethyl acetate to extract, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, the eluent is ethyl acetate, to obtain The compound monoamide carboxylate methyl ester represented by formula II provided by the invention (specifically compound 14 in the above reaction formula) (yield: 80%, ee 60.0%, Dicel-OJH).

14: solid, mp 93.0°C IR (KBr) v 3419, (CONH ₂ ), 1732, 1683 cm ^-1 ; ¹ H NMR (300 MHz, 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); ¹³ C 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 ₈ H ₁₄ N ₂ O ₃ : C, 51.60; H, 7.58; N, 15.04; Found: C, 51.14; H, 7.40 ; N, 15.38..

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula II.

Embodiment 11: the compound 15 shown in preparation formula II

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) wash the thalline into 150 milliliters of Erlenmeyer flat-bottomed flasks with threaded openings, disperse and shake well, and put into a shaker for activation at 30°C for 30 minutes, then add 1 mmol (197 mg) of the formula IV3 substrate in one shot Amides were placed in a shaker at 30° C. and 200 rpm to carry out catalytic hydrolysis reaction. The entire reaction was monitored by TLC, and the reaction was stopped after 7 days of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove bacteria, and the filter residue was washed with 20 ml of water for three times.

2) Freeze-dry the filtrate obtained in step 1) (-50 to -60°C), infiltrate the residue with 2 ml of methanol, and then drop into diazomethane diethyl ether solution with a concentration of 0.5mol/L at -15°C Carry out the methyl esterification reaction under the hood for 12 hours, add a small amount of water after the end, then use ethyl acetate to extract, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, the eluent is ethyl acetate, to obtain The compound monoamide carboxylate methyl ester represented by formula II provided by the invention (specifically compound 15 in the above reaction formula) (yield: 60%, ee 95.2%, Dicel-ADH).

15: solid; mp 72.0-74.0°C; IR(KBr)v 3364, 3292, 3265, 1686, 1650cm ^-1 ; [α] ²⁵ _D -64° (c 0.5, CHCl ₃ ); ¹ H 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=12.9 Hz, 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); ¹³ C 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 ₁₁ H ₁₃ BrN ₂ O: C, 49.09; H, 4.87; N, 10.41. Found: C, 49.10; H, 4.68; N, 10.23.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula II.

Embodiment 12: the compound 16 shown in preparation formula II

Its reaction formula is as follows

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) Wash the bacterium into a 150 ml Erlenmeyer flat-bottomed flask with screw top, disperse and shake well, put it in a shaker and activate it at 30°C for 30 minutes, then add 1 mg dissolved in 2.5 ml dimethyl sulfoxide at one time Mole (247 mg) of the formula IV4 substrate amide was put into a shaker at 30° C. and 200 rpm to carry out catalytic hydrolysis reaction. The entire reaction was monitored by TLC, and the reaction was stopped after 4 days of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove bacteria, and the filter residue was washed three times with 20 ml of water successively.

2) Freeze-dry the filtrate obtained in step 1) (-50 to -60°C), infiltrate the residue with 2 ml of methanol, and then drop into diazomethane diethyl ether solution with a concentration of 0.5mol/L at -15°C Carry out the methyl esterification reaction under the hood for 12 hours, add a small amount of water after the end, then use ethyl acetate to extract, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, the eluent is ethyl acetate, to obtain The present invention provides the compound monoamide carboxylate methyl ester represented by formula II (specifically compound 16 in the above reaction formula) (yield: 15%, ee > 99.5%, Dicel-ADH).

16: oil; ¹ HNMR (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); ¹³ CNMR (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. For C ₁₄ H ₁₈ N ₂ O ₃ : C, 64.10; H, 6.92; N, 10.68. Found: C, 64.25; H, 6.87; N, 10.92.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula II.

Embodiment 13: the compound 17 shown in preparation formula II

Its reaction formula is as follows:

The specific implementation method is:

1) Take two grams of Rhodococcus erythropolis AJ270 cells with a wet weight, thaw them at 30°C for 30 minutes, wash the cells with a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml) In an Erlenmeyer flat-bottomed flask with a threaded opening in milliliters, disperse and shake evenly and put it into a shaker for activation at 30°C for 30 minutes, then add 1 mmol (171 mg) 2,6-disubstituted hexahydropiperidine amide in one go, Put it into a shaker at 30° C. and 200 rpm to carry out the catalytic hydrolysis reaction. The whole reaction was monitored by TLC, and the reaction was stopped after 1 hour of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove the bacteria, and the filter residue was washed three times with 15 ml of water.

2) The filtrate obtained in step 1) was freeze-dried (-50 to -60°C), and the residue was stirred with 2 ml of N,N'-dimethylformamide, 1 ml of benzyl bromide, and 1.38 g of potassium carbonate at room temperature at 25°C for 24 Add a small amount of water after the esterification reaction in 1 hour, then use ethyl acetate to extract, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, and the eluent is ethyl acetate to obtain the present invention Benzyl monoamide carboxylate of the compound represented by formula II (specifically, compound 17 in the above reaction formula) (yield: 96%, ee > 99.5%, Dicel-ADH).

17: solid mp 105.0°C IR (KBr) v 3420 (CONH ₂ ), 1744, 1632 cm ^-1 ; ¹ H NMR (300 MHz, 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); ¹³ C 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 ₂₁ H ₂₄ N ₂ O ₃ , C, 71.57; H, 6.86; N, 7.95; Found: C, 71.58;

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula II.

Embodiment 14: the compound 18 shown in preparation formula II

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of Rhodococcus erythropolis AJ270 cells with a wet weight, thaw them at 30°C for 30 minutes, wash the cells with a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml) In an Erlenmeyer flat-bottomed flask with a threaded opening in milliliters, disperse and shake evenly and put it into a shaker for 30 minutes at 30° C., and then add 1 mmol (185 mg) of the formula IV5 amide in a shaker for 30 minutes. The catalytic hydrolysis reaction is carried out at 200 rpm. The whole reaction was monitored by TLC, and the reaction was stopped after 1 hour of reaction. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove the bacteria, and the filter residue was washed three times with 15 ml of water.

2) The filtrate obtained in step 1) was freeze-dried (-50 to -60°C), and the residue was stirred with 2 ml of N,N'-dimethylformamide, 1 ml of benzyl bromide, and 1.38 g of potassium carbonate at room temperature at 25°C for 24 Add a small amount of water after the esterification reaction in 1 hour, then use ethyl acetate to extract, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, and the eluent is ethyl acetate to obtain the present invention Compound monoamide carboxylate benzyl ester shown in formula II (specifically compound 18 in the above reaction formula) (yield: 74%, ee 67%).

18: oil IR (KBr) v 3410 (CONH ₂ ), 1737, 1670 cm ^-1 ; ¹ H NMR (300 MHz, 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); ¹³ C 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 %) 367.18 (M ^++1, 20%), 389.26 (M ⁺⁺ 23, 100%); Anal. Calcd. for C ₂₂ H ₂₆ N ₂ O ₃ C, 72.11; H, 7.15; N, 7.64; Found : C, 72.07; H, 7.10; N, 7.79.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is the compound shown in formula II.

Embodiment 15: Compound X' shown in the preparation formula II

Its reaction formula is as follows:

Specific implementation method:

1) Take two grams of wet weight of Rhodococcus erythropolis AJ270 cells (Institute of Chemistry, Chinese Academy of Sciences), thaw at 30°C for 30 minutes, and use a buffer solution of dipotassium hydrogen phosphate and potassium dihydrogen phosphate (0.1M, pH 7.0, 50ml ) wash the thalline into a 150 ml Erlenmeyer flat-bottomed flask with a threaded opening, disperse and shake well, and put it into a shaker for activation at 30°C for 30 minutes. Millimole (291 mg) of the substrate amide shown in formula IV6 was placed in a shaker at 30° C. and 200 rpm to carry out catalytic hydrolysis reaction. The whole reaction was monitored by high-performance liquid phase, and the reaction was stopped after 39 hours of reaction to obtain a reaction solution containing the compound shown in the product formula X. The resulting reaction solution was filtered through a layer of diatomaceous earth to remove the bacteria, and the filter residue was washed with 20 ml of water for three times. .

2) The filtrate obtained in step 1) was freeze-dried (-50 to -60°C), the residue was infiltrated with 2 ml of methanol, and then dripped with diazomethane diethyl ether solution with a concentration of 0.5mol/L at -15°C Carry out the esterification reaction for 12 hours, add a small amount of water after the end, then use ethyl acetate to extract, dry over anhydrous sodium sulfate, concentrate and use 100-200 mesh silica gel column chromatography, the eluent is ethyl acetate, and obtain monoamide Methyl carboxylate (specifically, x' in the above reaction formula) (yield: 39%, ee>99.5%, Dicel-ADH).

x: solid mp 198°C; ¹ HNMR (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); ¹³ CNMR(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, 1044, 159 , 1352, 1229cm ^-1 ; MS (ESI) m/z 293.34 (M++1, 10%), 315.29 (M ⁺⁺ 23, 100%). Anal.Calcd.for: C ₁₄ H ₁₆ N ₂ O ₅ C, 57.53; H, 5.52; N, 9.58; Found: C, 57.75; H, 5.51, N, 9.58.

x': solid mp 104°C; ¹ HNMR (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 Br 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 ₁₅ H ₁₈ N ₂ O ₅ C, 58.82; H, 5.92; N, 9.15 Found: C, 58.87; H, 5.99; N, 9.10.

It can be seen from the above that the structure of the above-mentioned compound is correct, and it is compound X' shown in formula II.

Embodiment 16: the antitumor activity of pseudonucleoside compounds shown in formula I

Tetrazolium [MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] is a dye that can accept hydrogen atoms. The dehydrogenase associated with NADP in the mitochondria of living cells can convert yellow MTT into insoluble blue-purple formazon in cells, while dead cells have no such function. After dissolving formazon with DMSO, measure the optical density value with a microplate reader at a certain wavelength, which can quantitatively measure the survival rate of cells. The experimental method is as follows: select the adherent tumor cells in the logarithmic growth phase, digest them with trypsin, prepare a cell suspension of 5000 cells/ml with RPMI1640 culture medium with 10% calf serum, and inoculate them in a 96-well culture plate. Inoculate 100ul per well and culture at 37°C, 5% CO2 for 24h.

Add 10ul of samples to the experimental group, the final volume of each hole is 200ul, and make up with 1640 culture medium. Culture at 37°C, 5% CO ₂ for 3 days.

Discard the supernatant, add 100ul freshly prepared 0.5mg/ml MTT serum-free culture solution to each well, and continue to incubate at 37°C for 4h. Discard the supernatant carefully, and add 200ulDMSO to dissolve the MTT formazon precipitate, mix with a micro-ultrasonic oscillator, and measure the optical density value at a wavelength of 544nm on a microplate reader.

Tumor cell growth inhibition rate=(OD _control -OD _experiment )/(OD _control -OD _blank )×100%

It has been measured that the pseudonucleoside compound 1 prepared in Example 1 has a growth inhibition rate of 1.96% for Bel-7402 (liver cancer cells), and a growth inhibition rate of 1.96% for A-549 (lung cancer cells). It can be seen from the above that the pseudonucleoside compound 1 prepared in Example 1 of the present invention has a certain inhibitory effect on the growth of Bel-7402 (liver cancer cells) and A-549 (lung cancer cells).

Claims (7)

1. pseudonucleoside compounds shown in formula I; In the formula I, n is 1 and m is 0; *Represents chirality, R or S configuration; X is Wherein, R ¹ is an alkyl group whose total number of carbon atoms is 1-5; Y is R ² is -H, -CH ₃ or -CH ₂ CH ₃ . 2. m is 0, Y is shown in formula I in a kind of preparation claim 1 X is The method for the pseudonucleoside compound is method a or method b, wherein said method a comprises the steps of: 1) react the compound described in formula _II ' with SOCl in N,N-dimethylformamide, and the compound shown in formula IIa1 is obtained after the reaction is completed; In the formula II', n is 1; * represents chirality, which is R or S configuration; R ² is H, -CH ₃ or -CH ₂ CH ₃ ; R ³ is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH=CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ ; 2) The compound shown in the formula IIa1 obtained in step 1) is refluxed with NaN _{and ZnBr} in a mixed solution composed of isopropanol and water, and the compound shown in the formula IIb1 is obtained after the reaction is completed; 3) Refluxing the compound shown in formula IIb1 obtained in step 2) with LiAlH _in tetrahydrofuran for reflux reaction, and the compound shown in formula IIc1 is obtained after the reaction is completed; 4) The compound shown in formula IIc1 obtained in step 3) is reacted with acetic anhydride, pyridine and 4-dimethylamino-pyridine in dichloromethane. After the reaction is completed, m is 0 and Y is X is A pseudonucleoside compound, that is, a compound shown in formula IId1; In the formula IIa1, formula IIb1, formula IIc1 and formula IId1, n is 1; * represents chirality, which is R or S configuration; R ² is -H, -CH ₃ or -CH ₂ CH ₃ ; X for R ¹ is an alkyl group whose total number of carbon atoms is 1-5; Described method b comprises the steps: 1) react the compound described in the formula III' with SOCl in DMF, _and the compound shown in the formula IIIa1 is obtained after the reaction is completed; In the formula III', n is 1; * represents chirality, which is R or S configuration; R ² is -H, -CH ₃ , -CH ₂ CH ₃ or -CH ₂ CH=CH ₂ ; R ³ is -NH ₂ , -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ , R ³ ' is -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH═CH ₂ , -OCH ₂ C ₆ H ₅ or -OCH ₂ C ₆ H ₅ OCH ₃ ; 2) The compound shown in the formula IIIa1 obtained in step 1) is refluxed with NaN _{and ZnBr} in a mixed solution composed of isopropanol and water, and the compound shown in the formula IIIb1 is obtained after the reaction is completed; 3) The compound shown in the formula IIIb1 obtained in step 2) is refluxed with _LiAlH in tetrahydrofuran, and the compound shown in the formula IIIc1 is obtained after the reaction is completed; 4) The compound shown in formula IIIc1 obtained in step 3) is reacted with acetic anhydride, pyridine and 4-dimethylamino-pyridine in dichloromethane, and after the reaction is completed, m is 0 and Y is X is The pseudonucleoside compound, that is, the compound shown in formula IIId1, In the formula IIIa1, formula IIIb1, formula IIIc1 and formula IIId1, n is 1; * represents chirality, which is R or S configuration; R ² is -H, -CH ₃ or -CH ₂ CH ₃ ; X for R ¹ is an alkyl group with a total of 1-5 carbon atoms. 3. The method according to claim 2, characterized in that: in the step 1) of the method a, the compound of the formula II', _SOCl and N, the ratio of the amount of N-dimethylformamide is 1.0-10.0mmol: 0.1-2mL: 1-100mL; in the reaction step, the temperature is -10°C-50°C, and the time is 0.5-24 hours; In the step 2), the dosage ratio of the compound represented by the formula IIa1, NaN ₃ , ZnBr ₂ and the mixed solution composed of isopropanol and water is 0.1-10.0 mmol: 0.2-20 mmol: 0.1-10 mmol: 1 -10mL; in the mixed solution composed of isopropanol and water, the volume ratio of isopropanol to water is 1-10:0.1-100; in the reflux reaction step, the time is 1-24 hours; In the step 3), the dosage ratio of the compound represented by the formula IIb1, LiAlH ₄ and tetrahydrofuran is 0.1-10mmol: 0.5-50mmol: 1mL-20mL; in the reflux reaction step, the time is 6-48 hours; In the step 4), the dosage ratio of the compound represented by the formula IIc1, acetic anhydride, pyridine, 4-dimethylaminopyridine and dichloromethane is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL ; The reaction temperature is 0-50°C, and the time is 6-48 hours; In step 1) of the method b, the dosage ratio of the compound of the formula III', SOCl ₂ and N,N-dimethylformamide is 1.0-10.0mmol: 0.1-2mL: 1-100mL; In the reaction step, the temperature is -10°C-50°C, and the time is 0.5-24 hours; In the step 2), the dosage ratio of the compound represented by the formula IIIa1, NaN ₃ , ZnBr ₂ and the mixed solution composed of isopropanol and water is 0.1-10.0 mmol: 0.2-20 mmol: 0.1-10 mmol: 1 -10mL; in the mixed solution composed of isopropanol and water, the volume ratio of isopropanol to water is 1-10:0.1-100; in the reflux reaction step, the time is 1-24 hours; In the step 3), the dosage ratio of the compound represented by the formula IIIb1, LiAlH ₄ and tetrahydrofuran is 0.1-10mmol: 0.5-50mmol: 1mL-20mL; in the reflux reaction step, the time is 6-48 hours; In the step 4), the dosage ratio of the compound represented by the formula IIIc1, acetic anhydride, pyridine, 4-dimethylaminopyridine and dichloromethane is 0.1-10mmol: 0.5-50mmol: 0.01-1mmol: 1mL-20mL ; The reaction temperature is 0-50°C, and the time is 6-48 hours. 4. take the pseudonucleoside compound shown in formula I described in claim 1 as the antitumor drug of active ingredient. 5. The antitumor drug according to claim 4, characterized in that: the tumor is liver cancer or lung cancer. 6. The application of pseudonucleoside compounds shown in formula I in claim 1 in the preparation of antitumor drugs. 7. The application according to claim 6, characterized in that: the tumor is liver cancer or lung cancer.