CN109305989B

CN109305989B - Phosphamide derivative and preparation method and application thereof

Info

Publication number: CN109305989B
Application number: CN201810740600.XA
Authority: CN
Inventors: 邱关鹏; 张晨; 罗新峰; 魏用刚; 严庞科; 郑伟
Original assignee: Sichuan Haisco Pharmaceutical Co Ltd
Current assignee: Sichuan Haisco Pharmaceutical Co Ltd
Priority date: 2017-07-28
Filing date: 2018-07-09
Publication date: 2021-02-26
Anticipated expiration: 2038-07-09
Also published as: CN109305989A

Abstract

The invention relates to a compound shown in a general formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, and application of the compound in preparing medicaments for preventing or treating diabetes, tumor, hyperkinetic movement disorder, motor neuron diseases, brain protection, diseases related to a central nervous system, immunosuppression, organ tissue or cell variant rejection inhibition, immunoregulation and/or inflammatory diseases, convulsion, epilepsy or stroke, wherein the structure of the compound shown in the general formula (I) is Q-L-R¹The radical definitions are consistent with the specification definitions.

Description

Phosphamide derivative and preparation method and application thereof

Technical Field

The invention relates to a phosphamide derivative, an intermediate and a preparation method thereof, and application of the phosphamide derivative in preparation of medicines.

Background

Some of the currently marketed or studied drugs have defects of certain physicochemical and biological properties, such as poor solubility, low stability, easy moisture absorption, bad smell, unsuitability for preparation, difficulty for passing through blood brain barrier, inability for oral administration, significant hepatic first pass effect, low oral bioavailability, high dosage and administration frequency, insufficient in vivo distribution for treatment, large food impact, small safety window, large gastrointestinal irritation, large tissue irritation, toxic reaction caused by non-target organ distribution, excessive metabolism or long-term retention, and the like. Therefore, it is necessary to develop a new technology to achieve the technical effects of improving the physicochemical properties of the drug, eliminating the unpleasant odor, improving the absorption, distribution, transportation and metabolic processes of the drug in vivo, improving the bioavailability, improving the selectivity of the drug on the target site, reducing the toxic and side effects of the drug, prolonging the action time and the like.

Disclosure of Invention

The invention discovers that the physical and chemical properties of the medicine, such as the property, the stability, the fat solubility, the P-gp substrate property and the like of the medicine can be changed by introducing the phosphoramide group, so that the characteristic of the absorption and metabolism distribution in vivo is changed. After the phosphoryl modified drug enters into the body, water is interpreted under the action of in-vivo hydrolase to release the original drug. The existence time of the medicine in vivo can be prolonged by controlling the hydrolysis rate of the phosphoramide medicine, and the purpose of improving the specific action of the medicine on a target site for administration can be achieved by the characteristics of the distribution of hydrolase and the like.

The invention provides a compound shown in a general formula (I) and a stereoisomer or pharmaceutically acceptable salt thereof,

Q-L-R¹ (I)

wherein: when Q is selected from

When the temperature of the water is higher than the set temperature,

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R^q1is selected from H or C_1-4An alkyl group;

R¹is selected from

R^1a、R^1c、R^1eEach independently selected from H or C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group, a carboxyl group,said alkyl is optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^L1selected from H, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

R^L2each independently selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Alkoxy, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

n is selected from 1,2 or 3; m is selected from 0,1, 2,3 or 4; p is selected from 0 or 1;

when Q is selected from

When, L is selected from a bond; r^q2、R^q3、R^q4、R^q5Each independently selected from isopropyl or cyclopropylethyl;

L₃is selected from-O-or-OCH₂O-；

R¹、R^q6Each independently selected from H,

Provided that R is¹、R^q6Cannot be simultaneously H;

R^1a、R^1c、R^1eeach independently selected from H or C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^1b、R^1d、R^1fis selected from C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

when Q is selected from

When, L is selected from a bond;

R³selected from phenyl or tert-butyloxy;

R⁴selected from H or acetyl;

L₄selected from each independent bond or

Wherein L is₄Is connected with Q on the left and with R on the right¹Connecting;

w is selected from-OC (═ O) O-, -OC (═ O) -or-C (═ O) O-;

R²each independently selected from H,

R^L2selected from H, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

R^L3each independently selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Alkoxy, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from H,F、Cl、Br、I、C_1-4alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

n is selected from 0,1 or 2;

m is selected from 0,1 or 2;

when Q is selected from

When the current is over;

l is selected from the group consisting of a bond, R¹Is selected from

R^2a、R^2c、R^2eEach independently selected from H or C_1-6Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^2b、R^2d、R^2feach independently selected from C_1-10Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^q7is selected from H or C_1-6Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

when Q is selected from

When, L is selected from a bond;

R¹is selected from

R^1a、R^1c、R^1eEach independently selected from H or C_1-6Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^1b、R^1d、R^1feach independently selected from C_1-6Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy.

When Q is selected from

When L is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R¹is selected from

R^1b、R^1d、R^1feach independently selected from C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^L3、R^L4or R^L5Each independently selected from H or C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

with the proviso that the compound of formula (I) is not of the structure shown below:

in a preferred embodiment of the present invention, a compound represented by the general formula (I) and a stereoisomer or a pharmaceutically acceptable salt thereof, wherein:

when Q is selected from

When L is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R^q1is selected from H or C_1-4An alkyl group;

R¹is selected from

R^1a、R^1c、R^1eEach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

R^L1is selected from H or C_1-4An alkyl group;

R^L2each independently selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4An alkoxy group;

when Q is selected from

When the temperature of the water is higher than the set temperature,

R^q2、R^q3、R^q4、R^q5each independently selected from isopropyl or cyclopropylethyl;

L₃is selected from-O-or-OCH₂O-；

R¹、R^q6Each independently selected from H,

Provided that R is¹、R^q6Cannot be simultaneously H;

R^1a、R^1c、R^1eeach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

when Q is selected from

When, L is selected from a bond;

R³selected from phenyl or tert-butyloxy;

R⁴selected from H or acetyl;

L₄each independently selected from the group consisting of a bond or

w is selected from-OC (═ O) O-, -OC (═ O) -or-C (═ O) O-;

R¹、R²each independently selected from H,

Provided that R is¹、R²Cannot be simultaneously H;

R^1a、R^1c、R^1eeach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

when Q is selected from

When the current is over;

l is selected from the group consisting of a bond, R¹Is selected from

R^2a、R^2c、R^2eEach independently selected from H or C_1-6An alkyl group;

R^2b、R^2d、R^2feach independently selected from C_1-10An alkyl group;

R^q7is selected from H or C_1-6An alkyl group;

when Q is selected from

When the current is over;

l is selected from a bond;

R¹is selected from

R^1a、R^1c、R^1eEach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

when Q is selected from

When the current is over;

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R¹is selected from

R^1a、R^1c、R^1eEach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

R^L3、R^L4or R^L5Each independently selected from H or C_1-4An alkyl group.

when Q is selected from

When the temperature of the water is higher than the set temperature,

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R^L1selected from H or methyl;

R^L2each independently selected from H, F, Cl, Br, I, methyl or methoxy;

n is 1; m is selected from 0,1, 2,3 or 4; p is 1;

R¹is selected from

R^1a、R^1c、R^1eEach independently selected from H, methyl, ethyl, propyl or isopropyl;

R^1b、R^1d、R^1feach independently selected from methyl, ethyl, propyl or isopropyl;

when Q is selected from

When the current is over;

L₃is selected from-O-or-OCH₂O-；

R¹、R^q6Each independently selected from H,

Provided that R is¹、R^q6Cannot be simultaneously H;

R^1b、R^1d、R^1feach independently selected from methyl, ethyl, propyl, isopropyl, butyl or tert-butyl;

when Q is selected from

When the temperature of the water is higher than the set temperature,

l is selected from a bond;

R³selected from phenyl or tert-butyloxy;

R⁴selected from H or acetyl;

L₄each independently selected from the group consisting of a bond or

w is selected from-OC (═ O) O-, -OC (═ O) -or-C (═ O) O-;

R¹、R²each independently selected from H,

Provided that R is¹、R²Cannot be simultaneously H;

R^1a、R^1c、R^1eeach independently selected from H, methyl, ethyl or isopropyl;

R^1b、R^1d、R^1feach independently selected from methyl, ethyl or isopropyl;

R^L2is selected from H;

R^L3each independently selected from H, F, Cl, Br, I, methyl, ethyl, methoxy or ethoxy;

when Q is selected from

When the current is over;

l is selected from the group consisting of a bond, R¹Is selected from

R^q7Selected from H, methyl, ethyl, propyl or isopropyl;

R^2a、R^2c、R^2eeach independently selected from H, methyl, ethyl or isopropyl;

R^2b、R^2d、R^2feach independently selected from methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, pentyl or hexyl; when Q is selected from

When the current is over;

l is selected from a bond;

R¹is selected from

R^1b、R^1d、R^1feach independently selected from methyl, ethyl or isopropyl;

when Q is selected from

When the current is over;

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R¹is selected from

R^L3、R^L4or R^L5Each independently selected from H, methyl, ethyl, propyl or isopropyl.

The invention relates to a compound shown in a general formula (I) and a stereoisomer or pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following structures:

the invention provides a pharmaceutical composition, which contains a therapeutically effective dose of a compound shown in a general formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and an excipient.

The invention also relates to application of the compound of the general formula (I) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, or the pharmaceutical composition in preparing medicines for preventing or treating diabetes, tumor resistance, hyperkinetic movement disorder, motor neuron diseases, brain protection, diseases related to the central nervous system, immunosuppression, organ tissue or cell foreign body rejection inhibition, immune regulation and/or inflammatory diseases, convulsion, epilepsy or stroke.

In a preferred embodiment of the invention, the diabetes is selected from type II diabetes, the hyperkinetic movement disorder is selected from chorea, huntington's chorea, tourette's syndrome, tardive dyskinesia or unilateral tic movement, the tumor is selected from prostate cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer, liver cancer, breast cancer, esophageal cancer, gastric cancer, colorectal cancer, cholangiocarcinoma, nasopharyngeal cancer, testicular cancer, lymphatic cancer, head and neck cancer, squamous cell carcinoma, colorectal cancer, leukemia, brain cancer, neck tumor and liver cancer, hematological cancer or bladder cancer, the motor neuron disease is selected from amyotrophic lateral sclerosis, and the central nervous system related disease is selected from multiple sclerosis.

The invention provides a novel metformin derivative and a stereoisomer or pharmaceutically acceptable salt thereof.

The invention relates to a compound shown in a general formula (I-a) and a stereoisomer or pharmaceutically acceptable salt thereof,

Q-L-R¹ (I-a)

wherein:

q is selected from

R¹Is selected from

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group; saidOptionally further substituted by 0 to 4 alkyl groups selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

n is selected from 1,2 or 3; m is selected from 0,1, 2,3 or 4; p is selected from 0 or 1.

In a preferred embodiment of the present invention, the compound of formula (I) and stereoisomers or pharmaceutically acceptable salts thereof, wherein:

R^1a、R^1c、R^1eeach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

R^L1is selected from H or C_1-4An alkyl group;

R^L2each independently selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4An alkoxy group.

In a preferred embodiment of the invention, the compound is represented by the general formula (I-a), and a stereoisomer or pharmaceutically acceptable salt thereof, wherein

R^1a、R^1c、R^1eEach independently selected from H and methylEthyl, propyl or isopropyl;

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R^L1selected from H or methyl; r^L2Each independently selected from H, F, Cl, Br, I, methyl or methoxy;

n is 1; m is selected from 0,1, 2,3 or 4; p is 1.

In a preferred embodiment of the present invention, the compound of formula (I-a) and stereoisomers or pharmaceutically acceptable salts thereof, wherein the compound is selected from one of the following structures:

the invention relates to a pharmaceutical composition, which contains a therapeutically effective dose of the compound of the general formula (I-a) and a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient.

The invention also relates to a compound with the general formula (I-a) and a stereoisomer or pharmaceutically acceptable salt thereof, and application of a composition of the compound in preparation of medicines for preventing or treating diabetes.

A preferred embodiment of the present invention, wherein said diabetes is type II diabetes.

The invention provides a compound shown in a general formula (I-b) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof,

R^q1is selected from H or C_1-4An alkyl group;

R²is selected from

R^2a、R^2c、R^2eEach independently selected from H or C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^2b、R^2d、R^2feach independently selected from C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R³selected from H, F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

R⁴selected from H, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

n is selected from 1,2 or 3;

m is selected from 0,1, 2,3 or 4.

In a preferred embodiment of the present invention, a compound represented by the general formula (I-b) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof:

R^q1selected from H, methyl or ethyl;

R²is selected from

R^2a、R^2c、R^2eSelected from H, methyl, ethyl or isopropyl;

R^2b、R^2d、R^2fselected from methyl, ethyl or isopropyl;

R³selected from H, F, Cl, Br, I, methyl, ethyl, trifluoromethyl, methoxy, ethoxy or cyclopropyl;

R³is selected from H;

n is selected from 1,2 or 3;

m is selected from 0,1, 2,3 or 4.

The compounds of the present invention of the general formula (I-b) include, but are not limited to, the following compounds:

the invention also relates to a pharmaceutical composition, which contains a therapeutically effective dose of the compound of the general formula (I-b) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug thereof, and a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.

The invention also relates to an application of the compound shown in the general formula (I-b) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, or the pharmaceutical composition in preparing a medicament for treating cancer, preferably an application in preparing a medicament for treating one or more of prostate cancer, breast cancer or bladder cancer.

The invention provides a compound shown in a general formula (I-c) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof,

R¹is selected from

R^1b、R^1d、R^1feach independently selected from C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

l is selected from the group consisting of a bond or

Wherein, when L is not a bond, R³Directly connected with oxygen on a benzene ring;

w is selected from- (═ O) O-or-O (═ O) -;

R^L3each independently selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Alkoxy, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

n is selected from 0,1 or 2;

m is selected from 0,1 or 2;

with the proviso that the compound of the formula (I-c) cannot be

In a preferred embodiment of the present invention, a compound represented by the general formula (I-c) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,

R¹is selected from

R^1b、R^1d、R^1feach independently selected from methyl, ethyl or isopropyl;

l is selected from the group consisting of a bond or

w is selected from- (═ O) O-or-O (═ O) -;

R^L2is selected from H;

n is selected from 0,1 or 2;

m is selected from 0,1 or 2;

with the proviso that the compound of the formula (I-c) cannot be

The compounds to which the present invention relates include, but are not limited to, the following:

the invention also relates to a pharmaceutical composition, which contains a therapeutically effective dose of the compound of the general formula (I-c) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug thereof, and a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.

The invention also relates to an application of the compound shown in the general formula (I-c) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, or an application of the pharmaceutical composition in preparation of medicines for treating cancers, preferably, in preparation of medicines for treating one or more of pancreatic cancer, lung cancer, ovarian cancer, cervical cancer, liver cancer, breast cancer, esophageal cancer, gastric cancer, colorectal cancer, bile duct cancer, nasopharyngeal cancer, testicular cancer, lymphatic disease, head and neck cancer, blood cancer or bladder cancer.

The invention relates to a compound shown in a general formula (I-d) and a stereoisomer or pharmaceutically acceptable salt thereof,

Q-L-R¹ (I-d)

wherein:

q is selected from

R¹Is selected from

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R^L1selected from H, C_1-4Alkyl, aryl, heteroaryl, and heteroaryl,C_1-4Alkoxy or C_3-6Cycloalkyl, said alkyl, alkoxy or cycloalkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl radical, C_1-4Alkoxy or C_3-6Cycloalkyl, substituted with a substituent;

with the proviso that the compounds of the formula (I-d) are not

In a preferred embodiment of the present invention, the compound of the general formula (I-d) and a stereoisomer or a pharmaceutically acceptable salt thereof are provided, wherein

R^1a、R^1c、R^1eEach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

R^L1is selected from H or C_1-4An alkyl group;

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

n is 1; m is selected from 0,1, 2,3 or 4; p is 1.

In a preferred embodiment of the present invention, the compound of formula (I-d) and stereoisomers or pharmaceutically acceptable salts thereof, wherein the compound is selected from one of the following structures:

the invention relates to a pharmaceutical composition, which contains a therapeutically effective dose of a compound shown in general formula (I-d) or a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient.

The invention also relates to a compound shown in the general formula (I-d) and a stereoisomer or pharmaceutically acceptable salt thereof, and application of the composition shown in the general formula (I-d) in preparing brain protection medicaments.

The invention relates to a compound shown in a general formula (I-e) and a stereoisomer or pharmaceutically acceptable salt thereof,

Q-L-R¹ (I-e)

wherein:

q is selected from

R¹Is selected from

R^1aIs selected from H or C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^1bis selected from C_1-4An alkyl group; said alkyl is optionally further substituted by 0 to 4 substituents selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

l is selected from the group consisting of a bond or

with the proviso that the compound of formula (I) is not one of the following structures:

in a preferred embodiment of the present invention, the compound of the general formula (I-e) and a stereoisomer or a pharmaceutically acceptable salt thereof are provided, wherein

R^1aEach independently selected from H or C_1-4An alkyl group;

R^1beach independently selected from C_1-4An alkyl group;

R^L1selected from H, C_1-4An alkyl group;

R^1aEach independently selected from H, methyl, ethyl, propyl or isopropyl;

R^1beach independently selected from methyl, ethyl, propyl or isopropyl;

R^L1selected from H or methyl;

R^L2each independently selected from H, F, Cl, Br, methyl, methoxy, ethyl or ethoxy.

In a preferred embodiment of the present invention, the compound of formula (I-e) and stereoisomers or pharmaceutically acceptable salts thereof, wherein the compound is selected from one of the following structures:

the invention relates to a pharmaceutical composition, which contains a therapeutically effective dose of a compound shown as a general formula (I-e) or a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient.

The invention also relates to a compound shown in the general formula (I-e) and a stereoisomer or pharmaceutically acceptable salt thereof, and application of the composition shown in the general formula (I-e) in preparing a medicament for preventing or treating diseases related to the central nervous system.

A preferred embodiment of the present invention, wherein the central nervous system related disease is multiple sclerosis.

The invention provides a compound shown in a general formula (I-f) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof,

l is selected from a bond or-OCH₂O-；

R⁵、R^q6Each independently selected from H,

Provided that R is⁵、R^q6Cannot be simultaneously H;

R^5a、R^5c、R^5eeach independently selected from H or C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^5b、R^5d、R^5fis selected from C_1-4Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

provided that the compound of formula (I-f) is not one of the following structures;

in a preferred embodiment of the present invention, a compound represented by the general formula (I-f) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,

l is selected from a bond or-OCH₂O-；

R⁵、R^q6Each independently selected from H,

Provided that R is⁵、R^q6Cannot be simultaneously H;

R^5a、R^5c、R^5eeach independently selected from H, methyl, ethyl or isopropyl;

R^5b、R^5d、R^5feach independently selected from methyl, ethyl, propyl, isopropyl, butyl or tert-butyl;

the compounds of the present invention of the general formula (I-f) include, but are not limited to, the following compounds:

the invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I-f) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, in combination with a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.

The invention also relates to application of the compound shown in the general formula (I-f) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a cocrystal or a prodrug thereof, or the pharmaceutical composition in preparation of medicines for treating convulsion, epilepsy or cerebral apoplexy.

The present invention also relates to a method for treating convulsion, epilepsy or stroke, which comprises administering a compound of formula (I-f) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, and a pharmaceutical composition thereof.

The invention provides a compound shown in a general formula (I-g) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof,

R¹、R²each independently selected from H,

R³selected from phenyl or tert-butyloxy;

R⁴selected from H or acetyl;

l is selected from the group consisting of a bond or

Wherein, when L is not a bond, R¹、R²Directly linked to a phenyl group;

w is selected from-OC (═ O) O-, -OC (═ O) -or-C (═ O) O-;

n is selected from 0,1 or 2;

m is selected from 0,1 or 2.

In a preferred embodiment of the present invention, a compound represented by the general formula (I-g) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,

R¹、R²each independently selected from H,

R^1b、R^1d、R^1feach independently selected from methyl, ethyl or isopropyl;

R³selected from phenyl or tert-butyloxy;

R⁴selected from H or acetyl;

l is selected from the group consisting of a bond or

Wherein, when L is not a bond, R¹、R²Directly linked to a phenyl group;

L₁is selected from-OC (═ O) O-, -OC (═ O) -or-C (═ O) O-;

R^L2is selected from H;

n is selected from 0,1 or 2;

m is selected from 0,1 or 2.

The compounds of the present invention of the general formula (I-g) include, but are not limited to, the following compounds:

the invention also relates to a pharmaceutical composition, which comprises a therapeutically effective dose of the compound of the general formula (I-g) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, cocrystal or prodrug thereof, and a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.

The invention also relates to an application of the compound shown in the general formula (I-g) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, or the pharmaceutical composition in preparing a medicament for treating cancers, preferably in preparing a medicament for treating one or more of ovarian cancer, lung cancer, breast cancer, prostatic cancer, gastric cancer, squamous cell carcinoma, bladder cancer, colorectal cancer, leukemia, brain cancer, neck tumor and liver cancer.

The invention provides a compound shown in a general formula (I-h) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof,

R²is selected from

with the proviso that the compounds of the formula (I) are not

In a preferred embodiment of the present invention, a compound represented by the general formula (I-h) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,

R¹selected from H, methyl, ethyl, propyl or isopropyl;

R²is selected from

R^2b、R^2d、R^2feach independently selected from methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, pentyl or hexyl;

with the proviso that the compounds of the formula (I-h) are not

The compounds of the present invention of the general formula (I-h) include, but are not limited to, the following compounds:

the invention also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I-h) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, in combination with a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.

The invention also relates to application of the compound shown in the general formula (I-h) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, or the pharmaceutical composition in preparing medicines for immunosuppression, prevention or treatment of rejection reaction caused by organ, tissue or cell allorejection, prevention or treatment of immune regulation and/or inflammatory diseases.

The invention provides a compound shown in a general formula (I-I) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof,

R¹is selected from

In a preferred embodiment of the present invention, a compound represented by the general formula (I-I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof,

R^1b、R^1d、R^1feach independently selected from methyl, ethyl or isopropyl.

The compounds of the present invention of the general formula (I-I) include, but are not limited to, the following compounds:

the invention also relates to a pharmaceutical composition, which contains a therapeutically effective dose of the compound of the general formula (I-I) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug thereof, and a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.

The invention also relates to an application of the compound shown in the general formula (I-I) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof, or the pharmaceutical composition in preparing a medicament for resisting tumors, wherein the tumors are preferably solid tumors, more preferably liver cancer.

The invention provides a compound shown in a general formula (I-j) or a stereoisomer, a hydrate, a metabolite, a solvate, a pharmaceutically acceptable salt, a eutectic crystal or a prodrug thereof,

R¹is selected from

R^a、R^c、R^eEach independently selected from H or C_1-6Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

R^b、R^d、R^feach independently selected from C_1-6Alkyl, said alkyl being optionally further substituted by 0 to 4 substituents selected from F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy;

with the proviso that the compounds of the formula (I-j) are not

In a preferred embodiment of the present invention, a compound represented by the general formula (I-j) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, wherein:

R¹is selected from

R^a、R^c、R^eEach independently selected from H, methyl, ethyl or isopropyl;

R^b、R^d、R^feach independently selected from methyl, ethyl or isopropyl;

with the proviso that the compounds of the formula (I-j) are not

The compounds of the present invention of the general formula (I-j) include, but are not limited to, the following compounds:

the invention also relates to a pharmaceutical composition, which contains a therapeutically effective dose of the compound of the general formula (I-j) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, eutectic or prodrug thereof, and a pharmaceutically acceptable carrier, diluent, adjuvant, vehicle or excipient; the composition may further comprise one or more additional therapeutic agents.

The invention also relates to the use of a compound of formula (I-j) or a stereoisomer, hydrate, metabolite, solvate, pharmaceutically acceptable salt, co-crystal or prodrug thereof, or a pharmaceutical composition thereof, for the manufacture of a medicament for hyperkinetic movement disorders, preferably for the manufacture of a medicament for the treatment of chorea, Huntington's disease, tourette's syndrome, tardive dyskinesia or unilateral tic disorders.

The invention relates to a compound shown in a general formula (I-k) and a stereoisomer or pharmaceutically acceptable salt thereof,

Q-L-R¹ (I-k)

wherein:

q is selected from

R¹Is selected from

l is selected from the group consisting of a bond or

Wherein L is connected to Q on the left and R on the right¹Connecting;

R^L3、R^L4or R^L5Each independently selected from H or C_1-4Alkyl radical ofOptionally further substituted by 0 to 4 alkyl groups selected from H, F, Cl, Br, I, C_1-4Alkyl or C_1-4Substituted by a substituent of alkoxy.

In a preferred embodiment of the invention, the compound is represented by the general formula (I-k), a stereoisomer or a pharmaceutically acceptable salt thereof, wherein

R^1a、R^1c、R^1eEach independently selected from H or C_1-4An alkyl group;

R^1b、R^1d、R^1feach independently selected from C_1-4An alkyl group;

R^L3、R^L4or R^L5Each independently selected from H or C_1-4An alkyl group.

In a preferred embodiment of the present invention, the compound of formula (I-k) and stereoisomers or pharmaceutically acceptable salts thereof, wherein the compound is selected from one of the following structures:

the invention relates to a pharmaceutical composition, which contains a therapeutically effective dose of a compound shown as a general formula (I-k) or a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient.

The invention also relates to a compound shown in the general formula (I-k) and a stereoisomer or pharmaceutically acceptable salt thereof, and application of a composition of the compound shown in the general formula (I-k) in preparation of a medicament for treating motor neuron diseases.

In a preferred embodiment of the present invention, the motor neuron disease is amyotrophic lateral sclerosis.

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

Carbon, hydrogen, oxygen, sulfur, nitrogen or halogen referred to in the groups and compounds of the invention all include isotopes thereof, and carbon, hydrogen, oxygen, sulfur, nitrogen or halogen referred to in the groups and compounds of the invention are optionally further replaced by one or more of their corresponding isotopes, wherein isotopes of carbon include isotopes of carbon¹²C、¹³C and¹⁴c, isotopes of hydrogen including protium (H), deuterium (D, also known as deuterium), tritium (T, also known as deuterium), and isotopes of oxygen including¹⁶O、¹⁷O and¹⁸isotopes of O, sulfur including³²S、³³S、³⁴S and³⁶isotopes of S, nitrogen include¹⁴N and¹⁵isotopes of N, F¹⁹Isotopes of F, chlorine including³⁵Cl and³⁷cl, isotopes of bromine including⁷⁹Br and⁸¹Br。

"optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where the event or circumstance occurs or does not. Such as: "alkyl optionally substituted with F" means that the alkyl group may, but need not, be substituted with F, and the description includes the case where the alkyl group is substituted with F and the case where the alkyl group is not substituted with F.

"pharmaceutical composition" means a mixture of one or more compounds described herein or a physiologically/pharmaceutically acceptable salt thereof with other ingredients, wherein the other ingredients comprise physiologically/pharmaceutically acceptable carriers and excipients.

"carrier" refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

"excipient" refers to an inert substance added to a pharmaceutical composition to further depend on the administration of the compound. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and different types of starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like.

"prodrug" refers to a compound that can be converted under physiological conditions or by solvolysis to a compound of the invention that is biologically active. Prodrugs of the invention are prepared by modifying functional groups in compounds of the invention, which modifications may be removed by routine manipulation or in vivo, to yield the parent compound.

"stereoisomers" refers to isomers resulting from the different arrangement of atoms in a molecule, including cis, trans isomers, enantiomers and conformational isomers.

An "effective dose" refers to an amount of a compound that elicits a physiological or medical response in a tissue, system, or subject that is sought, including an amount of the compound that, when administered to a subject, is sufficient to prevent the onset of, or alleviate to some extent, one or more symptoms of the condition or disorder being treated.

Detailed Description

The following detailed description is provided for the purpose of illustrating the embodiments and the advantageous effects thereof, and is not intended to limit the scope of the present disclosure.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) and/or Mass Spectrometry (MS), and Supercritical Fluid Chromatography (SFC) resolves the chiral structure.

NMR shift (. delta.) of 10^-6The units in (ppm) are given.

NMR was measured using a (Bruker ADVANCE III 400) nuclear magnetic spectrometer using deuterated dimethyl sulfoxide (DMSO-d)₆) Deuterated chloroform (CDCl)₃) Deuterated methanol (CD)₃OD), internal standard Tetramethylsilane (TMS), 1HNMR information is tabulated in the following format: chemical shifts (multiplet (s, singlet; d, doublet; t, triplet; q, quartet; Y, YHeavy peaks; m, multiplet), number of protons).

MS was measured (Agilent 6120B (ESI)).

HPLC was carried out using an Agilent 1260DAD high pressure liquid chromatograph (Zorba x SB-C18100 x 4.6.6 mm).

The thin layer chromatography silica gel plate adopts HSGF254 of tobacco yellow sea or GF254 of Qingdao, the specification of the silica gel plate used by Thin Layer Chromatography (TLC) is 0.15 mm-0.20 mm, and the specification of the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm.

The column chromatography generally uses 200-300 mesh silica gel of the Tibet Huanghai silica gel as a carrier.

The known starting materials of the present invention can be synthesized by methods known in the art or purchased from Chengdu Kelong chemical industry, Chengdu Xiong chemical industry, Chengdu chemical industry, Aister (Chengdu) pharmaceutical technology GmbH, Shaoshan far chemical technology (Shanghai) GmbH, pharmaceutical industry GmbH of the national medicine group, Bailinghi Technology GmbH, etc.

Intermediate 1

Ethyl (2S) -2- [ [ [4- (hydroxymethyl) phenoxy ] - [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl ] amino ] propionate (intermediate 1)

isopropyl(2S)-2-[[[4-(hydroxymethyl)phenoxy]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate

Dissolving phosphorus oxychloride (1.54g, 10mmol) in 100ml dichloromethane, reducing the temperature to-78 ℃ under the protection of nitrogen, dropwise adding 20ml of a mixed tetrahydrofuran solution of 4-hydroxybenzyl alcohol 1a (3.56g, 10mmol) and triethylamine (2.03g, 20mmol), continuing stirring for 30min after dropwise adding, adding L-alanine isopropyl ester hydrochloride (3.37g, 20.1mmol), adding triethylamine (3.05g, 30.1mmol), and naturally heating to room temperature for 2 h. After the reaction was completed, the reaction mixture was washed once with 150ml of a saturated aqueous solution of sodium dihydrogenphosphate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography with ethyl acetate/petroleum ether of 0: 1to 1:1 as eluent. The title compound, intermediate 1, was obtained as a colorless oil, (1.5g, 35% yield).

¹H NMR(400MHz,CDCl₃)δ7.29(d,2H),7.19(dd,2H),5.08–4.90(m,3H),4.63(s,2H),3.99(dt,2H),3.48(d,1H),1.37(t,6H),1.25–1.22(m,12H).

³¹P NMR(162MHz,CDCl₃)δ9.40(s,1H).

LC-MS:431.1[M+1]。

Intermediate 2

Isopropyl (2S) -2- [ [ [4- (hydroxymethyl) phenoxy ] - (methoxymethyl) phosphoryl ] amino ] propanoate (intermediate 2)

isopropyl(2S)-2-[[[4-(hydroxymethyl)phenoxy]-(methoxymethyl)phosphoryl]amino]propanoate

(methoxymethyl) phosphonyl dichloride (3.00g, 18.4mmol) was dissolved in 100mL of dry methylene chloride, and a mixture of triethylamine (7.45g, 73.6mmol) and L-alanine isopropyl ester hydrochloride (3.09g, 18.4mmol) was added dropwise under nitrogen at-30 ℃ to react for 30 minutes after completion of the addition, 4- (hydroxymethyl) phenol 1a (3.43g, 27.6mmol) was added thereto and reacted at room temperature for 2 hours. 50mL of water was added, and the mixture was extracted with dichloromethane (100 mL. times.2), washed once with a saturated aqueous solution of sodium dihydrogenphosphate (50mL), washed once with a saturated brine (50mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent methanol: dichloromethane (v/v) ═ 0:100 to 1:50) to give the title compound, isointermediate 2, as a pale yellow oil (3.60g, 56.6% yield).

¹H NMR(400MHz,CDCl₃)δ7.34(d,2H),7.25-7.19(m,2H),4.99(m,1H),4.56(d,2H),4.17-4.04(m,1H),3.87-3.72(m,2H),3.53(t,1H),3.46(m,3H),1.60(d,1H),1.32(d,3H),1.26-1.19(m,6H).

LC-MS m/z＝346.1[M+1]。

Intermediate 3

Isopropyl (2S) -2- [ [ [4- (hydroxymethyl) -2-methoxy-phenoxy ] - (methoxymethyl) phosphono ] amino ] propanoate

isopropyl(2S)-2-[[[4-(hydroxymethyl)-2-methoxy-phenoxy]-(methoxymethyl)phosphoryl]amino]propanoate

(methoxymethyl) phosphonyl dichloride (3.2g, 19.6mmol) is dissolved in 30mL of dry dichloromethane, L-alanine isopropyl ester hydrochloride (3.29g, 19.6mmol) is added at-30 ℃ under the protection of nitrogen, triethylamine (7.95g, 78.6mmol) is slowly added, the reaction is continued for 1 hour, 3-hydroxy-4-methoxybenzyl alcohol 2a (3.03g, 19.6mmol) is added, and the mixture is naturally warmed to room temperature for reaction for 2 hours. Water (40mL) was added, liquid separation was performed, the aqueous phase was extracted with dichloromethane (40mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether: ethyl acetate (v/v) ═ 1: 1). The title compound, intermediate 3, was obtained as a pale yellow liquid (2.0g, 27% yield).

¹H NMR(400MHz,DMSO)δ7.27(d,1H),7.06-6.98(m,2H),5.50-5.32(m,1H),5.14-5.06(m,1H),4.90-4.80(m,1H),4.40(d,2H),3.96-3.84(m,1H),3.80-3.68(m,5H),3.37(dd,3H),1.22-1.14(m,9H).

31P NMR(162MHz,DMSO)δ25.80,25.08.

LC-MS(m/z)＝376.1[M+1]。

Intermediate 4

Isopropyl (2S) -2- [ [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (2,3,4,5, 6-pentafluorophenoxy) phosphoryl ] amino ] propionate (intermediate 4)

isopropyl(2S)-2-[[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-(2,3,4,5,6-pentafluorophenoxy)phosphoryl]amino]propanoate。

Pentafluorophenol (5g,27.17mmol) was dissolved in dichloromethane (200mL), cooled to-78 deg.C, phosphorus oxychloride (4.57g,29.88mol) was added, and triethylamine (3g,29.66mmol) was added dropwise. After stirring at this temperature for 20 minutes, L-alanine isopropyl ester hydrochloride (10g,59.52mmol) and triethylamine (12g,118.66mmol) were added and stirring was continued for 30min at room temperature. The solvent was removed under reduced pressure, methyl tert-butyl ether (100mL) was added, the filtrate was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)20/1 to 4/1) to give intermediate 4as a pale yellow solid (5g, 37.53% yield).

LC-MS m/z＝491.2[M+1].

Example 1

Isopropyl (2S) -2- [ [ [ [ N- (N, N-dimethylamidino) amidine ] amino ] - (methoxymethyl) phosphoryl ] amino ] propanoate (Compound 1)

Isopropyl(2S)-2-[[[[N-(N,N-dimethylcarbamimidoyl)carbamimidoyl]amino]-(methoxymethyl)phosphoryl]amino]propanoate.

The first step is as follows: isopropyl (2S) -2- [ [ methoxymethyl- (4-nitrophenoxy) phosphoryl ] amino ] propionate (1B)

isopropyl(2S)-2-[[methoxymethyl-(4-nitrophenoxy)phosphoryl]amino]propanoate。

Compound 1A (2g,14.38mmol) and (methoxymethyl) phosphoryl dichloride (2.34g,14.38mmol) are dissolved in dichloromethane (50mL) at-20 deg.C, triethylamine (1.45g,14.38mmol) is slowly added dropwise to the mixture, after stirring at this temperature for 20min, L-alanine isopropyl ester hydrochloride (2.42g,14.39mmol) and triethylamine (2.91g,28.78mmol) are added in this order, and stirring is continued for 30min while naturally warming to room temperature. Dichloromethane (50mL) was added, the mixture was washed with saturated sodium dihydrogen phosphate solution and brine in this order, the organic layer was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 10/1-4/1) to give compound 1B as a pale yellow viscous semisolid (3g, 57.90% yield).

LC-MS(m/z)：359.2[M-1]。

The second step is that:

isopropyl(2S)-2-[[[[N-(N,N-dimethylcarbamimidoyl)carbamimidoyl]amino]-(methoxymethyl)phosphoryl]amino]propanoate

A mixture of metformin (0.8g,6.19mmol) and acetonitrile (50mL) was placed in a three-necked flask, NaHMDS (2mol/L,4.6mL) was added dropwise under ice bath, the temperature was raised to room temperature and stirring was continued for 30min, and then the temperature was lowered to 0 ℃. To the mixture was added a solution of 1B (4.44g,12.3mmol) in tetrahydrofuran (10mL), the mixture was naturally warmed to room temperature, dichloromethane (100mL) was added, the mixture was washed with saturated brine (50mL × 2), the organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (methanol/dichloromethane (v/v) ═ 100/1 to 20/1) to obtain compound 1 as a pale yellow oil (0.17g, yield 7.84%).

¹H NMR(400MHz,DMSO--d₆)δ8.30(s,1H),6.08(s,2H),5.01–4.74(m,1H),4.20–3.97(m,1H),3.91–3.69(m,1H),3.56–3.35(m,5H),3.11–2.72(m,6H),1.29–1.26(m,3H),1.24–1.20(m,6H)。

³¹P NMR(162MHz,DMSO-d₆)δ21.668,21.171。

LC-MS(m/z)：351.1[M+1]。

Example 2

Isopropyl (2S) -2- [ [ [ [ N- (dimethylformamidine) amidine ] amino ] - [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl ] amino ] propionate (Compound 2)

Isopropyl(2S)-2-[[[[N-(N,N-dimethylcarbamimidoyl)carbamimidoyl]amino]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate

The first step is as follows:

isopropyl (2S) -2- [ [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (2,3,4,5, 6-pentafluorophenoxy) phosphoryl ] amino ] propionate (2B)

Compound 2A (5g,27.17mmol) was dissolved in dichloromethane (200mL), cooled to-78 deg.C, phosphorus oxychloride (4.57g,29.88mol) was added, and triethylamine (3g,29.66mmol) was added dropwise. After stirring at this temperature for 20 minutes, L-alanine isopropyl ester hydrochloride (10g,59.52mmol) and triethylamine (12g,118.66mmol) were added and stirring was continued for 30min at room temperature. The solvent was removed under reduced pressure, methyl tert-butyl ether (100mL) was added, the filtrate was filtered, and the filtrate was concentrated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 20/1 to 4/1) to give compound 2B as a pale yellow solid (5g, yield 37.53%).

LC-MS m/z＝491.2[M+1]。

The second step is that:

isopropyl(2S)-2-[[[[N-(N,N-dimethylcarbamimidoyl)carbamimidoyl]amino]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate。

A mixture of metformin (0.8g,6.20mmol) and acetonitrile (40mL) was added to a three-necked flask, NaHMDS (2mol/L,6.2mL) was added dropwise under ice bath, the temperature was raised to room temperature and stirring continued for 30min, after which the temperature was lowered to 0 ℃. To the mixture was added a solution of 2B (6g,12.22mol) in tetrahydrofuran (10mL), the mixture was allowed to warm to room temperature, dichloromethane (100mL) was added, the mixture was washed with saturated brine (50mL × 2), the organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (methanol/dichloromethane (v/v) ═ 100/1 to 20/1) to give compound 2 as a pale yellow oil (0.22g, 8.15% yield).

¹H NMR(400MHz,DMSO-d₆)δ8.14(s,1H),6.69(s,1H),4.97–4.76(m,2H),3.99–3.57(m,4H),2.88(d,6H),1.23-1.17(m,18H).

³¹P NMR(162MHz,DMSO-d₆)δ15.35。

LC-MS m/z＝436.2[M+1]。

Example 3

Isopropyl (2S) -2- [ [ [ (1S) -2- [4- [ (4-fluoro-2-methyl-1H-indol-5-yl) oxy ] -5-methyl-pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] oxy-1-methyl-ethoxy ] - (methoxymethyl) phosphoryl ] amino ] propanoate (Compound 3)

isopropyl(2S)-2-[[[(1S)-2-[4-[(4-fluoro-2-methyl-1H-indol-5-yl)oxy]-5-methyl-pyrrolo[2,1-f][1,2,4]triazin-6-yl]oxy-1-methyl-ethoxy]-(methoxymethyl)phosphoryl]amino]propanoate

Bulbonib 1A (0.25g,0.67mmol) was dissolved in tetrahydrofuran (10mL), tert-butyl magnesium chloride (1mol/L,1.3mL) was added dropwise in ice bath, the temperature was raised to room temperature, stirring was continued for 30min, and then the temperature was lowered to 0 ℃. To the mixture was added a solution of 1B (0.48g,1.33mol) in tetrahydrofuran (5mL), the mixture was naturally warmed to room temperature, dichloromethane (20mL) was added, the mixture was washed with saturated brine (20mLx 2), the organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (methanol/dichloromethane (v/v)100/1 to 20/1) to obtain compound 3 as a yellow solid (0.17g, yield 42.89%).

¹H NMR(400MHz,DMSO-d₆)δ11.32(s,1H),7.93(m,2H),7.14(d,1H),6.99(dd,1H),6.24(s,1H),5.18(m,1H),4.90-4.85(m,2H),4.07(m,2H),3.83(m,1H),3.62-3.55(m,2H),3.32(m,3H),2.41(dd,6H),1.35(m,3H),1.25(m,3H),1.18(m,6H).

31P NMR(162MHz,DMSO-d6)δ26.04,25.02.

LC-MS m/z＝592.2[M+1]

Example 4

Isopropyl (2S) -2- [ [ [ (1S) -2- [4- [ (4-fluoro-2-methyl-1H-indol-5-yl) oxy ] -5-methyl-pyrrolo [2,1-f ] [1,2,4] triazin-6-yl ] oxy-1-methyl-ethoxy ] - [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl ] amino ] propionate (Compound 4)

isopropyl(2S)-2-[[[(1S)-2-[4-[(4-fluoro-2-methyl-1H-indol-5-yl)oxy]-5-methyl-pyrrolo[2,1-f][1,2,4]triazin-6-yl]oxy-1-methyl-ethoxy]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate；

Bulborne (1A) (0.3g,0.81mmol) was dissolved in tetrahydrofuran (10mL), tert-butyl magnesium chloride (1mol/L,1.6mL) was added dropwise in ice bath, and after warming to room temperature and stirring continued for 30min, the temperature was lowered to 0 ℃. To the mixture was added a tetrahydrofuran (5mL) solution of intermediate 4(0.8g,1.63mol), the mixture was naturally warmed to room temperature, dichloromethane (20mL) was added, the mixture was washed with saturated brine (20mLx 2), the organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (methanol/dichloromethane (v/v)100/1 to 20/1) to obtain compound 2 as a white solid (0.35g, 63.86% yield).

1H NMR(400MHz,DMSO-d6)δ11.32(s,1H),7.93(m,2H),7.14(d,1H),6.98(dd,1H),6.24(s,1H),4.89-4.83(m,2H),4.67(m,2H),4.55(m,1H),4.08(d,2H),3.76(m,2H),2.41(m,6H),1.26(m,9H),1.20-1.14(m,12H).

31P NMR(162MHz,DMSO-d6)δ13.20。

Example 5

(2S) -isopropyl 2- [ [ [ (2R,3R,11bR) -3-isobutyl-9, 10-dimethoxy-2, 3,4,6,7,11 b-hexahydro-1H-benzo [ a ] quinolizin-2-yl ] oxy- (methoxymethyl) phosphoryl ] amino ] propionate (Compound 5)

isopropyl(2S)-2-[[[(2R,3R,11bR)-3-isobutyl-9,10-dimethoxy-2,3,4,6,7,11b-hexahydro-1H-benzo[a]quinolizin-2-yl]oxy-(methoxymethyl)phosphoryl]amino]propanoate

The first step is as follows: (2R,3R,11bR) -3-isobutyl-9, 10-dimethoxy-2, 3,4,6,7, 11B-hexahydro-1H-pyridin [2,1-a ] isoquinolin-2-ol (5B)

(2R,3R,11bR)-3-isobutyl-9,10-dimethoxy-2,3,4,6,7,11b-hexahydro-1H-pyrido[2,1-a]isoquinolin-2-ol

(3R,11bR) tetrabenazine (5A) (10.00g,31.51mmol) was dissolved in dry tetrahydrofuran (100mL), cooled at-20 deg.C with stirring, and BH was added dropwise₃/CH₃SCH₃Solution (2M) (31.5mL), stirred at-20 ℃ for 2h, added with ammonia (110mL), warmed to 35 ℃ for reaction for 2h, reacted at room temperature overnight, and the stirring was stopped. Ethyl acetate (50mL) was added, the layers were separated, and the organic layer was washed successively with saturated aqueous ammonium chloride (20 mL. times.1) and saturated brine (20 mL. times.1). After drying over anhydrous sodium sulfate, concentration under reduced pressure, and column chromatography, light yellow solid 5B (5.00g, yield 49.70%) was obtained.

LCMS m/z＝320.3[M+1].

¹H NMR(400MHz,d₆-DMSO)δ6.71(s,1H),6.63(s,1H),3.70(d,1H),4.56(d,6H),3.16-3.08(m,1H),2.98(d,1H),2.82-2.83(m,3H),2.51-2.41(m,3H),2.31-2.24(m,1H),1.84(t,1H),1.66-1.65(m,2H),1.48-1.43(m,1H),1.27-1.18(m,1H),0.89(d,3H),0.85(d,3H)。

The second step is that: (2S) -isopropyl 2- [ [ [ (2R,3R,11bR) -3-isobutyl-9, 10-dimethoxy-2, 3,4,6,7,11 b-hexahydro-1H-benzo [ a ] quinolizin-2-yl ] oxy- (methoxymethyl) phosphoryl ] amino ] propionate (Compound 5)

(methoxymethyl) phosphoryl dichloride (5C) (0.74g,4.51mmol) was dissolved in dichloromethane (20mL), 5B (0.96g,3.01mmol) was added in portions under nitrogen protection at 0 ℃, then triethylamine (2.10mL,15.00 mmol) was added dropwise, after stirring at 0 ℃ for 60min, L-alanine isopropyl ester hydrochloride (0.76g, 4.51mmol) was added, the reaction was naturally warmed to room temperature for 4h, then stirring was stopped, washing was performed with water (10mL × 2) and saturated aqueous sodium chloride solution (10mL × 1) in this order, drying was performed with sodium sulfate, and after concentration under reduced pressure, purification by silica gel column chromatography (anhydrous petroleum ether/ethyl acetate (v/v): 1/2) was performed to obtain compound 5 as a yellow solid (0.47g, yield 28.9%).

LC-MS m/z＝541.3[M+1].

¹H NMR(400MHz,CDCl₃)δ6.69(s,1H),6.57(s,1H),5.08-5.02(m,1H),4.28-4.20(m,1H),3.84(d,J＝2.4Hz,6H),3.70(d,J＝8.3Hz,2H),3.44(s,3H),3.35-3.20(m,1H),3.23-3.20(m,1H),3.11-3.00(m,4H),2.83-2.78(m,1H),2.67-2.45(m,3H),2.10-2.06(m,1H),1.76-1.66(m,1H),1.53-1.47(m,1H),1.44-1.39(m,3H),1.28-1.25(m,6H),1.14-1.07(m,2H),0.99-0.90(m,6H).

³¹P NMR(162MHz,CDCl₃)δ25.21.

Example 6

(2S) -Ethyl 2- [ [ [ (2R,3R,11bR) -3-isobutyl-9, 10-dimethoxy-2, 3,4,6,7,11 b-hexahydro-1H-benzo [ a ] quinolizin-2-yl ] oxy- (methoxymethyl) phosphono ] amino ] propionate (Compound 6)

ethyl(2S)-2-[[[(2R,3R,11bR)-3-isobutyl-9,10-dimethoxy-2,3,4,6,7,11b-hexahydro-1H-benzo[a]quinolizin-2-yl]oxy-(methoxymethyl)phosphoryl]amino]propanoate

(methoxymethyl) phosphoryl dichloride (5C) (0.653g,4.01mmol) was dissolved in dichloromethane (20mL), 5B (0.64g,2.00mmol) was added in portions under nitrogen protection at 0 ℃, triethylamine (2.20mL,16.00mmol) was then added dropwise, after stirring at 0 ℃ for 60min, L-alanine ethyl ester hydrochloride (0.62g, 4.01mmol) was added, the reaction was naturally warmed to room temperature for 4h, then the reaction was stopped, washed with water (10mL × 2), washed with saturated sodium chloride (10mL × 1), dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography for purification (petroleum ether/ethyl acetate (v/v) ═ 1/2) after concentration under reduced pressure to obtain compound 6 as a yellow solid (0.27g, yield 25.6%).

LC-MS m/z＝527.3[M+1].

¹H NMR(400MHz,CDCl₃)δ6.68(m,1H),6.66(s,1H),4.30-3.98(m,3H),3.89(s,2H),3.84(s,4H),3.73-3.69(m,2H),3.53-3.20(m,6H),3.12-3.00(m,3H),2.78-2.73(m,1H),2.66-2.62(m,1H),2.50-2.44(m,1H),2.09-2.02(m,3H),1.99-1.86(m,1H),1.71-1.53(m,2H),1.44-1.42(m,3H),1.30-1.17(m,3H),0.99-0.88(m,6H).

³¹P NMR(162MHz,CDCl₃)δ25.44.

Example 7

Isopropyl (2S) -2- [ [ [ (3R,11bR) -3-isobutyl-9, 10-dimethoxy-2, 3,4,6,7,11 b-hexahydro-1H-benzo [ a ] quinolizin-2-yl ] oxy- [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl ] amino ] propionate (Compound 7)

isopropyl(2S)-2-[[[(3R,11bR)-3-isobutyl-9,10-dimethoxy-2,3,4,6,7,11b-hexahydro-1H-benzo[a]quinolizin-2-yl]oxy-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate

5B (1.50g,4.70mmol) was dissolved in dry tetrahydrofuran (20ml), added dropwise with nitrogen protection, t-BuMgBr in tetrahydrofuran (6.57ml,1.0M), heated to 50 ℃ for 30 minutes, added with isopropyl (2S) -2- [ [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (4-nitrophenyl) phosphoryl ] amino ] propionate (3.35g,7.51mmol), and heated to reflux for 6 hours. The reaction was quenched with 50ml of water, extracted with ethyl acetate (30 ml. times.3), the organic phases were combined, dried over sodium sulfate, filtered, concentrated, and purified by column chromatography to give compound 3(0.40g, yield: 13.60%).

LC-MS m/z＝626.4[M-1].

¹H NMR(400MHz,CDCl3)δ6.70(s,1H),6.56(s,1H),5.07-5.01(m,1H),4.97-4.91(m,1H),4.18-4.11(m,1H),3.98-3.92(m,1H),3.89(s,3H),3.84(s,3H),3.37-3.32(m,1H),3.24-3.19(m,1H),3.05-2.98(m,4H),2.80(br,1H),2.64(br,1H),2.47(br,1H),1.89(br,1H),1.76-1.63(m,2H),1.55-1.49(m,2H),1.41(d,3H),1.37(d,3H),1.27-1.24(m,7H),1.21(d,3H),1.15-1.08(m,4H),0.95-0.91(m,6H).

Example 8

Isopropyl (2S) -2- [ [ [4- [ [ [4- [3- [ 4-cyano-3- (trifluoromethyl) phenyl ] -5, 5-dimethyl-4-oxo-2-thioxo-imidazol-1-yl ] -2-fluoro-benzoyl ] -methyl-carbamoyl ] oxymethyl ] phenoxy ] - (methoxymethyl) phosphoryl ] amino ] propanoate (Compound 8)

isopropyl(2S)-2-[[[4-[[[4-[3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]-2-fluoro-benzoyl]-methyl-carbamoyl]oxymethyl]phenoxy]-(methoxymethyl)phosphoryl]amino]propanoate。

Dissolving compound 8A (2g,4.31mmol) in tetrahydrofuran (20mL), adding chlorotrimethylsilane (0.56g,5.15mmol) and triethylamine (0.52g,5.15mmol) in sequence under ice bath, heating the mixture to 60 ℃, stirring for 30min, cooling to room temperature, adding triphosgene (0.69g,2.33mmol), heating to 60 ℃, and stirring for 2 h. The solvent was removed under reduced pressure to give a yellow oil. In another three-necked flask, intermediate 2(1.50g,4.34mmol) was dissolved in tetrahydrofuran (10mL) and triethylamine (0.52g,5.15mmol) and 4-dimethylaminopyridine (0.53g,4.34mmol) were added. Under ice-bath conditions, a tetrahydrofuran solution (5mL) of the above yellow oil was slowly added dropwise. After the dripping is finished, slowly raising the temperature to the room temperature, and continuously stirring for reaction for 1 h. Dichloromethane (30mL) was added for extraction and washed successively with saturated sodium dihydrogen phosphate solution (20mL × 1) and sodium chloride solution (20mL × 1). The organic layer was dried over anhydrous sodium sulfate and concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1 to 1/1) to give compound 8 as a white solid (1g, yield 27.76%).

¹HNMR(400MHz,CDCl₃)δ8.04–7.92(m,2H),7.87(dd,1H),7.61(t,1H),7.25–7.13(m,5H),6.96(d,1H),5.21–4.81(m,3H),4.22–3.91(m,1H),3.88–3.66(m,2H),3.60–3.49(m,1H),3.47(d,1H),3.45–3.34(m,5H),1.59(s,6H),1.31(d,3H),1.26–1.18(m,6H).

³¹P NMR(162MHz,CDCl₃)δ24.23,23.28.

Example 9

Isopropyl (2S) -2- [ [ [4- [ [4- [3- [ 4-cyano-3- (trifluoromethyl) phenyl ] -5, 5-diethyl-4-oxo-2-thioxo-imidazol-1-yl ] -2-fluoro-benzoyl ] carbamoyloxymethyl ] phenoxy ] - (methoxymethyl) phosphoryl ] amino ] propanoate (Compound 9)

isopropyl(2S)-2-[[[4-[[4-[3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-2-thioxo-imidazolidin-1-yl]-2-fluoro-benzoyl]carbamoyloxymethyl]phenoxy]-(methoxymethyl)phosphoryl]amino]propanoate

Compound 9A (0.6g,1.33mmol) was dissolved in 1, 2-dichloroethane, oxalyl chloride (0.25g,1.97mmol) was added, the mixture was heated to reflux, stirred for 30min, cooled to room temperature, and concentrated under reduced pressure to give a yellow oil. In another three-necked flask, intermediate 2(0.46g,1.33mmol) was dissolved in tetrahydrofuran (10mL) and triethylamine (0.16g,1.58mmol) was added. Under ice-bath conditions, a tetrahydrofuran solution (5mL) of the above yellow oil was slowly added dropwise. After the dripping is finished, slowly raising the temperature to the room temperature, and continuously stirring for reaction for 1 h. Dichloromethane (30mL) was added for extraction, and the mixture was washed successively with saturated sodium dihydrogen phosphate solution (20 mL. times.1) and sodium chloride solution (20 mL. times.1). The organic layer was dried over anhydrous sodium sulfate and concentrated, and the residue was separated and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1 to 1/1) to give compound 9 as a white solid (300mg, yield 27.45%).

¹HNMR(400MHz,CDCl₃)δ8.50(d,1H),8.20(t,1H),8.07–7.88(m,2H),7.87–7.73(m,1H),7.38(d,2H),7.30(d,1H),7.21(dd,2H),5.23(s,2H),5.00(dd,1H),4.10(s,1H),3.89–3.71(m,2H),3.56(s,1H),3.46(d,3H),1.62(s,6H),1.33(d,3H),1.26–1.18(m,6H).

³¹P NMR(162MHz,CDCl₃)δ24.25,23.29.

Example 10

Isopropyl (2S) -2- [ [ [4- [ [1- [ (2R,4R,5R) -3, 3-difluoro-4-hydroxy-5- (hydroxymethyl) tetrahydrofuran-2-yl ] -2-oxo-pyrimidin-4-yl ] carbamoyloxymethyl ] phenoxy ] - [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl ] amino ] propanoate (compound 10)

isopropyl(2S)-2-[[[4-[[1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]-2-oxo-pyrimidin-4-yl]carbamoyloxymethyl]phenoxy]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate

The first step is as follows:

isopropyl (2S) -2- [ [ [4- [ [1- [ (2R,4R,5R) -3, 3-difluoro-4-trimethylsilyloxy-5- (trimethylsiloxymethyl) tetrahydrofuran-2-yl ] -2-oxo-pyrimidin-4-yl ] carbamoyloxymethyl ] phenoxy ] - [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl ] amino ] propionate (10B)

isopropyl(2S)-2-[[[4-[[1-[(2R,4R,5R)-3,3-difluoro-4-trimethylsilyloxy-5-(trimethylsilyloxymethyl)tetrahydrofuran-2-yl]-2-oxo-pyrimidin-4-yl]carbamoyloxymethyl]phenoxy]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate

Intermediate 1(3.0g, 6.97mmol) was dissolved in tetrahydrofuran (20ml), cooled to 0 ℃ under nitrogen, triphosgene (1.03g, 3.49mmol) was added, diisopropylethylamine (1.35g, 10.5mmol) was added and stirring in an ice bath continued for 30 minutes after addition was complete. Separately, 10A (2.84g, 6.97mmol) was dissolved in dichloromethane (20ml), 4-dimethylaminopyridine (1.27g, 10.4mmol) was added thereto, and the reaction mixture was slowly dropped into the mixture under ice bath, and then the mixture was naturally warmed to room temperature to react for 3 hours. Dichloromethane (30ml) and saturated sodium dihydrogen phosphate (100ml) were added, the mixture was separated, the organic layer was washed with saturated sodium chloride (100ml), dried over anhydrous sodium sulfate, and the crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) ═ 4: 1to 1:4) to give compound 10B as a white solid (1.00g, yield 16.6%).

The second step is that:

10B (1.0g, 1.2mmol) was dissolved in tetrahydrofuran (10ml), and an aqueous solution (2ml) of ammonium fluoride (1.0g, 27mmol) was added at room temperature, and the reaction was continued for 1 hour after the addition was completed. 10mL of water was added, extraction was performed with dichloromethane (20 mL. times.2), the organic layers were combined, washed with saturated sodium chloride (20 mL. times.1), dried over anhydrous sodium sulfate, and the resulting crude product was concentrated under reduced pressure and separated and purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate (v/v) ═ 1: 4. about. methanol: dichloromethane (v/v) ═ 1:19) to give compound 10 as a white solid (0.48g, yield 58%).

¹H NMR(400MHz,DMSO-d₆)δ10.93(s,1H),8.22(d,1H),7.38(d,2H),7.18(d,2H),7.10(d,1H),6.29(d,1H),6.16(t,1H),5.28(t,1H),5.24–5.09(m,4H),4.94–4.79(m,2H),4.27–4.12(m,1H),3.94–3.74(m,4H),3.68-3.62(m,1H),1.25(t,6H),1.20-1.12(m,12H).

³¹P NMR(162MHz,DMSO-d₆)δ10.67.

LC-MS(M/Z)＝720.3[M+1].

Example 11

Ethyl (2S) -2- [ [ methoxymethyl- [ [6- (trifluoromethoxy) -1, 3-benzothiazol-2-yl ] amino ] phosphoryl ] amino ] propionate (compound 11)

ethyl(2S)-2-[[methoxymethyl-[[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]amino]phosphoryl]amino]propanoate

(methoxymethyl) phosphoryl dichloride (1.25g,7.69mmol) was dissolved in 8mL of dichloromethane, a solution of 11A (1.50g,6.41mmol) and triethylamine (5.2g,51.24mmol) in dichloromethane (3mL) was added dropwise at-10 ℃ under nitrogen, and after dropping, the reaction was kept in an ice bath for 2h, and at this temperature, L-alanine ethyl ester (0.90g,7.69mmol) was added and the reaction was carried out at room temperature for 4 h. Washed with water (20mL), dried over anhydrous sodium sulfate for 0.5h, concentrated and the residue chromatographed on neutral alumina column (PE/EA ═ 1/1 → DCM/MeOH ═ 10/1) and separated by preparative HPLC to give ethyl (2S) -2- [ [ methoxymethyl- [ [6- (trifluoromethoxy) -1, 3-benzothiazol-2-yl ] amino ] phosphoryl ] amino ] propionate (compound 11) as a yellow oil (70mg, 2% yield).

The preparation conditions were as follows:

the instrument comprises the following steps: waters 2767;

column: xbridge C18, 19 × 250mm i.d.,5 μm.;

mobile phase: a for acetonitrile and B for 0.1% ammonium acetate in water; gradient: a is 60 to 80 percent;

flow rate: 12 ml/min;

back pressure: 1900 PSI;

column temperature: 30 ℃;

wavelength: 210 nm;

and (3) period: and 15min.

LCMS m/z＝442.1[M+1]。

Example 12

Isopropyl (2S) -2- [ [ methoxymethyl- [ [6- (trifluoromethoxy) -1, 3-benzothiazol-2-yl ] amino ] phosphoryl ] amino ] propanoate (Compound 2)

isopropyl(2S)-2-[[methoxymethyl-[[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]amino]phosphoryl]amino]propanoate

(methoxymethyl) phosphoryl dichloride (1.25g,7.69mmol) was dissolved in 8mL of dichloromethane, a solution of 11A (1.50g,6.41mmol) and triethylamine (5.2g,51.24mmol) in dichloromethane (3mL) was added dropwise at-10 ℃ under nitrogen, the ice bath was maintained for 2h after the addition, and L-isopropyl alanine (1.01g,7.69mmol) was added at this temperature and the reaction was carried out at room temperature for 4 h.

Washed with water (20mL), dried over anhydrous sodium sulfate for 0.5h, concentrated, and the residue purified by silica gel column chromatography (PE/EA ═ 1:1-0:1) and isolated by preparative HPLC to give compound 2 as a yellow oil (80mg, 3% yield).

The preparation conditions were as follows:

the instrument comprises the following steps: gilson GX-281;

column: xbridge C18, 30 × 150mm i.d.,5 μm.;

mobile phase: a for acetonitrile and B for 0.1% ammonium acetate in water; gradient: 30% -70% of A;

flow rate: 30 mL/min;

back pressure: 1000 PSI;

column temperature: 30 ℃;

wavelength: 210 nm;

and (3) period: and 15min.

LCMS m/z＝456.1[M+1]。

¹HNMR(400MHz,CDCl₃)：δ7.52-7.46(m,2H),7.21-7.19(m,1H),5.05-5.01(m,1H),4.16-4.14(m,1H),3.92-3.89(m,2H),3.87-3.84(m,1H),3.47(s,3H),1.47-1.43(m,3H),1.33-1.20(m,6H)。

Example 13

Isopropyl (2S) -2- [ [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - [ [6- (trifluoromethoxy) -1, 3-benzothiazol-2-yl ] amino ] phosphono ] amino ] propanoate (Compound 13)

isopropyl(2S)-2-[[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-[[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]amino]phosphoryl]amino]propanoate

Dissolving a compound phosphorus oxychloride (1.2g,7.6mmol) in dichloromethane (15mL), dropwise adding a solution of riluzole 11A (1.80g,7.6mmol) and triethylamine (1.08mL) in dichloromethane (10mL) at 0 ℃ under the protection of nitrogen, naturally raising the temperature to room temperature, continuously stirring for 1 hour, dropwise adding a solution of L-isopropyl alanine (2.7g,15.2mmol) in dichloromethane (5mL), slowly dropwise adding the rest triethylamine (4.4mL), and continuously stirring for 3 hours. The reaction mixture was washed with water (20mL), separated, the organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) ═ 1:1-0:1) to give compound 3 as a pale yellow oil (0.5g, yield 12.2%).

LC-MS m/z＝541.2[M+1]。

¹H NMR(400MHz,CDCl₃)δ9.46(s,1H),7.90(s,1H),7.54(s,1H),7.29-7.26(m,1H),4.99(brs,2H),4.90-4.76(m,2H),3.89-3.82(m,2H),1.28-1.26(d,6H),1.17-1.14(t,6H),1.12-1.08(m,6H)。

³¹P NMR(162MHz,CDCl₃)δ5.83。

Example 14

Ethyl (2S) -2- [ [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - [ [6- (trifluoromethoxy) -1, 3-benzothiazol-2-yl ] amino ] phosphono ] amino ] propanoate (compound 14)

ethyl(2S)-2-[[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-[[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]amino]phosphoryl]amino]propanoate

Dissolving a compound phosphorus oxychloride (1.2g,7.6mmol) in dichloromethane (15mL), dropwise adding a dichloromethane (10mL) solution of riluzole 11A (1.80g,7.6mmol) and triethylamine (1.08mL) at 0 ℃ under the protection of nitrogen, naturally raising the temperature to room temperature, continuously stirring for 1 hour after dropwise adding, dropwise adding a dichloromethane (5mL) solution of L-alanine ethyl isopropyl ester (2.1g,15.2mmol), slowly dropwise adding the rest triethylamine (4.4mL), and continuously stirring for 3 hours. The reaction mixture was washed with water (20mL), separated, the organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) ═ 1:1-0:1) to give compound 14 as a white solid (0.49g, yield 12.6%).

LC-MS m/z＝513.2[M+1]。

¹H NMR(400MHz,CDCl₃)δ9.46(s,1H),7.90(s,1H),7.54(s,1H),7.29-7.27(m,1H),5.02(brs,2H),4.07-3.95(m,4H),390-3.88(d,2H),1.29-1.28(d,5H),1.24(s,2H),1.17-1.08(m,5H)。

³¹P NMR(162MHz,CDCl₃)δ7.50。

Example 15

Isopropyl 2- [ [ [ (2-isopropoxy-2-oxo-ethyl) amino ] - [ [6- (trifluoromethoxy) -1, 3-benzothiazol-2-yl ] amino ] phosphoryl ] amino ] acetate (compound 15)

isopropyl2-[[[(2-isopropoxy-2-oxo-ethyl)amino]-[[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]amino]phosphoryl]amino]acetate

Dissolving a compound phosphorus oxychloride (1.2g,7.6mmol) in dichloromethane (15mL), dropwise adding a solution of riluzole (1.80g,7.6mmol) and triethylamine (1.08mL) in dichloromethane (10mL) at 0 ℃ under the protection of nitrogen, naturally raising the temperature to room temperature, continuously stirring for 1 hour, dropwise adding a solution of isopropyl glycinate (2.1g,15.2mmol) in dichloromethane (5mL), slowly dropwise adding the rest triethylamine (4.4mL), and continuously stirring for 3 hours. The reaction mixture was washed with water (20mL), separated, the organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) ═ 1:1-0:1) to give compound 15 as a pale yellow oil (0.49g, yield 12.6%).

LC-MS m/z＝541.2[M+1]。

¹H NMR(400MHz,CDCl₃)δ9.49(s,1H),7.90(s,1H),7.56-7.54(d,1H),7.29-7.26(m,1H),4.97(brs,2H),4.92-4.83(m,2H),3.65-3.60(m,4H),1.16-1.14(m,12H)。

³¹P NMR(162MHz,CDCl₃)δ8.53。

Example 16

Ethyl 2- [ [ [ (2-isopropoxy-2-oxo-ethyl) amino ] - [ [6- (trifluoromethoxy) -1, 3-benzothiazol-2-yl ] amino ] phosphoryl ] amino ] acetate (compound 16)

Ethyl 2-[[[(2-isopropoxy-2-oxo-ethyl)amino]-[[6-(trifluoromethoxy)-1,3-benzothiazol-2-yl]amino]phosphoryl]amino]acetate

Dissolving a compound phosphorus oxychloride (1.2g,7.6mmol) in dichloromethane (15mL), dropwise adding a dichloromethane (10mL) solution of riluzole 11A (1.80g,7.6mmol) and triethylamine (1.08mL) at 0 ℃ under the protection of nitrogen, naturally raising the temperature to room temperature, continuously stirring for 1 hour, dropwise adding a dichloromethane (5mL) solution of isopropyl glycinate (1.65g,15.2mmol), slowly dropwise adding the rest triethylamine (4.4mL), and continuously stirring for 3 hours. The reaction mixture was washed with water (20mL), separated, the organic phase was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) ═ 1:1-0:1) to give compound 16 as a white solid (0.45g, yield 12.2%).

LC-MS m/z＝485.1[M+1]。

¹H NMR(400MHz,CDCl₃)δ9.62(s,1H),7.90(s,1H),7.56-7.53(d,1H),7.29-7.26(m,1H),4.99(brs,2H),4..08-4.02(m,4H),3.68-3.63(m,4H),1.17-1.13(t,6H)。

³¹P NMR(162MHz,CDCl₃)δ10.52。

Example 17

Isopropyl (2S) -2- [ [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (5-methyl-2-phenyl-pyrazol-3-yl) oxy-phosphoryl ] amino ] propionate (Compound 17)

Isopropyl(2S)-2-[[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-(5-methyl-2-phenyl-pyrazol-3-yl)oxy-phosphoryl]amino]propanoate

Dissolving phosphorus oxychloride (4.4g, 29mmol) in tetrahydrofuran (50ml), cooling to 0 ℃ under the protection of nitrogen, adding 17A (5.0g, 29mmol), slowly dropwise adding triethylamine (3.5g, 34.8mmol), stirring for 2 hours after the addition is finished, adding L-alanine isopropyl ester hydrochloride (9.6g, 57mmol), dropwise adding triethylamine (5.2g, 52.2mmol), and naturally heating to room temperature for reaction for 2 hours after the addition is finished. The mixture was filtered, and the filter cake was washed with 10ml of ethyl acetate, and the filtrate was collected and concentrated under reduced pressure, and the resulting crude product was purified by a silica gel column (dichloromethane: ethyl acetate (v/v) ═ 1:0 to 9:1) to give compound 17 as a pale yellow oil (1.0g, yield 7.3%).

¹H NMR(400MHz,DMSO-d6)δ7.65-7.61(m,2H),7.49–7.41(m,2H),7.35–7.27(m,1H),6.01(s,1H),5.60-5.46(m,2H),4.90–4.77(m,2H),3.87–3.74(m,2H),2.17(s,3H),1.26-1.20(m,6H),1.16-1.10(m,12H)。

³¹P NMR(162MHz,DMSO-d6)δ11.04。

LC-MS(M/Z)＝481.2[M+1]。

Example 18

Isopropyl (2S) -2- [ [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - [4- [ (5-methyl-2-phenyl-pyrazol-3-yl) oxycarbonyloxymethyl ] phenoxy ] phosphoryl ] amino ] propionate (compound 18)

Isopropyl(2S)-2-[[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-[4-[(5-methyl-2-phenyl-pyrazol-3-yl)oxycarbonyloxymethyl]phenoxy]phosphoryl]amino]propanoate

Intermediate 1(2.47g, 5.74mmol) was dissolved in tetrahydrofuran (20ml), cooled to 0 ℃ under nitrogen, triphosgene (0.852g, 2.87mmol) was added, diisopropylethylamine (1.11g, 8.61mmol) was added and stirring in an ice bath continued for 30 minutes after addition was complete. 17A (1.00g, 5.74mmol) was dissolved in tetrahydrofuran (10ml), diisopropylethylamine (0.742g, 5.74mmol) was added thereto, and the reaction mixture was added dropwise in ice bath, and then allowed to warm to room temperature naturally for 2 hours. The mixture was filtered, and the filter cake was washed with 10ml of ethyl acetate, and the filtrate was collected and concentrated under reduced pressure, and the resulting crude product was purified by a silica gel column (dichloromethane: ethyl acetate (v/v) ═ 1:0 to 9:1) to give compound 18 as a pale yellow oil (0.600g, yield 16.6%).

¹H NMR(400MHz,DMSO-d6)δ7.51–7.43(m,4H),7.38–7.30(m,3H),7.19(d,2H),6.22(s,1H),5.27–5.13(m,4H),4.93–4.80(m,2H),3.91–3.76(m,2H),2.23(s,3H),1.29–1.21(m,6H),1.20–1.11(m,12H)。

³¹P NMR(162MHz,DMSO-d6)δ10.66。

LC-MS(M/Z)＝631.3[M+1]。

Example 19

Ethyl 2- [ [ [ (2-ethoxy-2-oxo-ethyl) amino ] - (5-methyl-2-phenyl-pyrazol-3-yl) oxy-phosphoryl ] amino ] acetate (compound 19)

Ethyl2-[[[(2-ethoxy-2-oxo-ethyl)amino]-(5-methyl-2-phenyl-pyrazol-3-yl)oxy-phosphoryl]amino]acetate

Phosphorus oxychloride (3.04g, 19.8mmol) is dissolved in tetrahydrofuran (50ml), cooled to 0 ℃ under nitrogen protection, 17A (3.45g, 19.8mmol) is added, triethylamine (2.40g, 23.7mmol) is slowly added dropwise, stirring is continued for 2 hours after the addition is finished, glycine ethyl ester hydrochloride (5.5g, 39.6mmol) is added dropwise, triethylamine (7.6g, 75.2mmol) is added dropwise, and the temperature is naturally raised to room temperature after the addition is finished for reaction for 2 hours. The filtrate was filtered, and the cake was washed with 10ml of ethyl acetate, and the filtrate was collected and concentrated under reduced pressure, and the resulting crude product was purified by a silica gel column (dichloromethane: ethyl acetate (v/v) ═ 1:0 to 9:1) to give the title compound (compound 19) as a pale yellow oil (1.2g, yield 14.3%).

¹H NMR(400MHz,CDCl₃)δ7.60-7.56(m,2H),7.46–7.39(m,2H),7.33-7.27(m,1H),6.02(s,1H),4.21–4.12(m,4H),3.80–3.59(m,6H),2.27(s,3H),1.25(t,6H)。

³¹P NMR(162MHz,CDCl₃)δ11.61。

LC-MS(M/Z)＝425.2[M+1]。

Example 20

Isopropyl (2S) -2- [ [ methoxymethyl- [4- [ (5-methyl-2-phenyl-pyrazol-3-yl) oxycarbonyloxymethyl ] phenoxy ] phosphoryl ] amino ] propanoate (Compound 20)

Isopropyl(2S)-2-[[methoxymethyl-[4-[(5-methyl-2-phenyl-pyrazol-3-yl)oxycarbonyloxymethyl]phenoxy]phosphoryl]amino]propanoate

Intermediate 2(1.90g, 5.74mmol) was dissolved in tetrahydrofuran (20ml), cooled to 0 ℃ under nitrogen, triphosgene (0.85g, 2.87mmol) was added, diisopropylethylamine (1.11g, 8.60mmol) was added and stirring in an ice bath continued for 30 minutes after addition was complete. 17A (1.00g, 5.74mmol) was dissolved in tetrahydrofuran (10ml), diisopropylethylamine (0.740g, 5.74mmol) was added thereto, and the reaction mixture was added dropwise in ice bath, and then allowed to warm to room temperature naturally for 2 hours. The mixture was filtered, and the filter cake was washed with 10ml of ethyl acetate, and the filtrate was collected and concentrated under reduced pressure, and the resulting crude product was purified by a silica gel column (dichloromethane: ethyl acetate (v/v) ═ 1:0 to 9:1) to give compound 20 as a pale yellow oil (0.800g, yield 25.5%).

¹H NMR(400MHz,DMSO-d6)δ7.51–7.43(m,4H),7.39–7.32(m,3H),7.24–7.15(m,2H),6.22(s,1H),5.74-5.64(m,1H),5.24(s,2H),4.88-4.80(m,1H),3.92-3.82(m,1H),3.80-3.74(m,2H),3.38(dd,3H),2.23(s,3H),1.20-1.12(m,9H).

³¹P NMR(162MHz,DMSO-d6)δ24.91,24.10。

LC-MS(M/Z)＝546.2[M+1]。

Example 21

Isopropyl (2S) -2- [ [ methoxymethyl- [ 2-methoxy-5- [ (5-methyl-2-phenyl-pyrazol-3-yl) oxycarbonyloxymethyl ] phenoxy ] phosphoryl ] amino ] propionate (compound 21)

isopropyl(2S)-2-[[methoxymethyl-[2-methoxy-5-[(5-methyl-2-phenyl-pyrazol-3-yl)oxycarbonyloxymethyl]phenoxy]phosphoryl]amino]propanoate

Intermediate 3(0.646g, 1.72mmol) was dissolved in tetrahydrofuran (10ml), cooled to 0 ℃ under nitrogen, triphosgene (0.256g, 0.86mmol) was added, diisopropylethylamine (0.333g, 2.58mmol) was added and stirring in an ice bath continued for 30 minutes after addition was complete. Further, 17A (0.300g, 1.72mmol) was dissolved in tetrahydrofuran (5ml), diisopropylethylamine (0.557g, 4.31mmol) was added thereto, and the reaction mixture was added dropwise under ice bath, and then allowed to warm to room temperature naturally for 2 hours.

LC-MS(M/Z)＝576.2[M+1]。

Example 22

O4- [ [3- [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxo-4-methoxy-phenyl ] methyl ] O1-methyl (E) -butenedioate (Compound 22)

O4-[[3-[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-(methoxymethyl)phosphoryl]oxy-4-methoxy-phenyl]methyl]O1-methyl(E)-but-2-enedioate

Dissolving monomethyl fumarate 22A (0.39g,3.00mmol) in 10mL of dichloromethane, adding two drops of N, N-dimethylformamide and oxalyl chloride (1mL), stirring for 1h, and concentrating under reduced pressure to remove excess oxalyl chloride; the residue was dissolved in dry dichloromethane (10mL), triethylamine and a solution of intermediate 3 in dichloromethane (5mL) were added and stirred at room temperature for 4 h. Diluting with water (20mL), separating the organic phase, extracting the aqueous phase with dichloromethane (20 mL. times.2), combining the organic phases, washing with saturated brine (40mL), drying over anhydrous sodium sulfate for 0.5h, concentrating under reduced pressure, and then performing column chromatography (petroleum ether/ethyl acetate ═ 1:1) to give a yellow oil, preparative isolation gave compound 22(0.32g, 22.0% yield)

The preparation conditions were as follows:

the instrument comprises the following steps: waters 2767;

column: xbridge C18, 19 × 250mm i.d.,5 μm.;

mobile phase: a for acetonitrile and B for 0.1% ammonium acetate in water; gradient: 50% -80% of A;

flow rate: 12 ml/min;

back pressure: 1900 PSI;

column temperature: 30 ℃;

wavelength: 210 nm;

and (3) period: LCMS M/z 488.2[ M +1 ] at 15min]⁺

¹HNMR(400MHz,CDCl₃)：δ7.36-7.33(m,1H),7.16-7.14(m,1H),6.93-6.91(m,1H),6.87(s,1H),5.14-5.13(m,1H),5.02-4.91(m,1H),4.14-4.03(m,1H),3.91-3.80(m,8H),3.52-3.49(m,3H),1.34-1.18(m,9H).

Example 23

(E) -4-methoxy-4-oxo-2-butenoic acid [ 2-fluoro-4- [ [ [ (1S) -2-isopropoxycarbonyl ] ethylamino ] - (methoxymethyl) phosphate ] benzyl ester (compound 23)

O4-[[2-fluoro-4-[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-(methoxymethyl)phosphoryl]oxy-phenyl]methyl]O1-methyl(E)-but-2-enedioate

The first step is as follows: (2S) -2- [ [ 3-fluoro-4- (hydroxymethylphenoxy) ] - (methoxymethyl) phosphinamido ] isopropyl propionate (23B)

(2S)-isopropyl-2-[[3-fluoro-4-(hydroxymethy)]phenoxy)(methoxymethyl)phosphoryl)amino]propanoate

Methoxymethylphosphoryl chloride (3.26g,20mmol) was dissolved in dichloromethane (30mL), cooled to-30 deg.C, a solution of the reagent isopropyl (2S) -2-aminopropyl acetate (2.62g,20mmol) in dichloromethane (10mL) was added dropwise, and after stirring for 1h, a suspension of triethylamine (8.09g, 80mmol) and 3-fluoro-4-hydroxymethylphenol (1.42g, 10mmol) in dichloromethane (10mL) was added, and the mixture was allowed to warm to room temperature and stirred for 3 h. Saturated aqueous sodium bicarbonate (20mL) and water (30mL) were added, the organic phase was separated, the aqueous phase was extracted with dichloromethane (50mL × 2), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate ═ 1:1to 1:5) to give orange-yellow liquid 23B (2.32g, yield 29.1%).

MS M/Z(ESI):364.1(M+1).

The second step is that: (E) -4-methoxy-4-oxo-2-butenoic acid [ 2-fluoro-4- [ [ [ (1S) -2-isopropoxycarbonyl ] ethylamino ] - (methoxymethyl) phosphate ] benzyl ester (compound 24)

Monomethyl fumarate 22A (0.65g,5mmol) was dissolved in dichloromethane (30mL), two drops of N, N-dimethylformamide and oxalyl chloride (1.5mL) were added, the mixture was stirred for 1 hour, and then concentrated under reduced pressure to remove excess oxalyl chloride; the residue was dissolved in dry dichloromethane (20mL), triethylamine (1.01g,10mmol) and a solution of 23B (1.82g, 5mmol) in dichloromethane (10mL) were added and the mixture was stirred at room temperature for 4 h. Water (20mL) was added for dilution, the organic phase was separated, the aqueous phase was extracted with dichloromethane (20mL × 2), the organic phases were combined, washed with saturated brine (40mL × 1), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (petroleum ether/ethyl acetate ═ 1:1) to give compound 23(0.205g, yield 8.6%) as a colorless liquid.

MS M/Z(ESI):474.1(M-1).

¹HNMR:(400MHz,CDCl₃):δ7.40-7.31(m,1H),7.05-7.01(m,2H),6.88(s,1H),5.25(s,1H),5.04-4.96(m,1H),4.59(s,1H),4.14-4.05(m,1H),3.86-3.75(m,3H),3.61-3.56(m,1H),3.49-3.44(m,3H),1.34(d,J＝7.0Hz,3H),1.25-1.21(m,6H).

Example 24:

methyl (E) -6- [4- [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxy-6-methoxy-7-methyl-3-oxo-1 hydro-isobenzofuran-5-yl ] -4-methyl-hex-4-enoate (compound 24)

methyl(E)-6-[4-[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-(methoxymethyl)phosphoryl]oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate

The first step is as follows: methyl (E) -6- (4-hydroxy-6-methoxy-7-methyl-3-oxo-1 hydro-isobenzofuran-5-yl) -4-methyl-hex-4-enoate (24B)

methyl(E)-6-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl)-4-methyl-hex-4-enoate

Compound 24A (1.5g,4.68mmol) and p-toluenesulfonic acid (0.81g,4.70mmol) were dissolved in methanol (20mL), heated to reflux and stirred for 2 h. The solvent was removed under reduced pressure, the residue was dissolved in ethyl acetate (50mL), washed successively with saturated sodium bicarbonate solution (50mL x 3) and brine (50mL x 1), and the organic layer was dried and concentrated to give compound 24B as a white solid (1.5g, 95.86% yield).

¹H NMR(400MHz,CDCl₃)δ7.67(s,1H),5.31–5.11(m,3H),3.76(s,3H),3.62(s,3H),3.38(d,2H),2.44–2.34(m,2H),2.35–2.24(m,2H),2.15(s,3H),1.80(s,3H).

The second step is that: methyl (E) -6- [4- [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl ] -4-methyl-hex-4-enoate (compound 24)

Compound 24B (0.5g,1.50mmol) was dissolved in dichloromethane (20mL), and (methoxymethyl) phosphonodichloride (0.25g,1.53mmol) was added under ice-bath conditions followed by triethylamine (0.5g,4.94mmol) and stirring at 0 ℃ for 20min followed by addition of L-alanine isopropyl ester hydrochloride (0.26g,1.55 mmol). After warming to room temperature and stirring for further 30min, dichloromethane (50mL) was added, and the mixture was washed with saturated sodium dihydrogen phosphate solution (50 mL. times.1) and saturated brine (50 mL. times.1) in this order. After the organic layer was dried and concentrated, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1 to 1/1) to give compound 24 as a pale yellow oil (500mg, yield 60.00%).

¹H NMR(400MHz,CDCl₃)δ5.26–5.09(m,3H),5.03-4.78(m,1H),4.69–4.55(m,1H),4.10–3.92(m,3H),3.92–3.73(m,4H),3.61(d,3H),3.56–3.41(m,4H),2.43–2.33(m,2H),2.33–2.24(m,2H),2.20(dd,3H),1.79(s,3H),1.37(d,2H),1.27–1.18(m,3H),1.11(d,2H),1.06(d,2H).

³¹P NMR(162MHz,CDCl₃)δ27.27,26.31.

LC-MS(m/z):556.2[M+1]

Example 25:

ethyl (E) -6- [4- [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl ] -4-methyl-hex-4-enoate (compound 25)

ethyl(E)-6-[4-[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]-(methoxymethyl)phosphoryl]oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate。

The first step is as follows:

ethyl (E) -6- (4-hydroxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl) -4-methyl-hex-4-enoate (2B)

ethyl(E)-6-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl)-4-methyl-hex-4-enoate

Compound 24A (1.5g,4.68mmol) and p-toluenesulfonic acid (0.81g,4.70mmol) were dissolved in ethanol (20mL), heated to reflux and stirred for 2 h. The solvent was removed under reduced pressure, the residue was dissolved in ethyl acetate (50mL), washed successively with saturated sodium bicarbonate solution (50mL x 3) and saturated brine (50mL x 1), and the organic layer was dried and concentrated to give compound 25B as a white solid (1.5g, 92.00% yield).

¹H NMR(400MHz,CDCl₃)δ7.67(s,1H),5.24(m,1H),5.26-5.19(s,2H),4.07(q,2H),3.76(s,3H),3.38(d,2H),2.44–2.34(m,2H),2.33–2.25(m,2H),2.15(s,3H),1.80(s,3H),1.21(t,3H).

The second step is that: ethyl (E) -6- [4- [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl ] -4-methyl-hex-4-enoate (compound 25)

Compound 25B (0.5g,1.44mmol) was dissolved in dichloromethane (20mL), and (methoxymethyl) phosphonodichloride (0.25g,1.53mmol) was added under ice-bath conditions followed by triethylamine (0.5g,4.94mmol) and stirring at 0 ℃ for 20min followed by addition of L-alanine isopropyl ester hydrochloride (0.26g,1.55 mmol). After warming to room temperature and stirring for further 30min, dichloromethane (50mL) was added, and the mixture was washed with saturated sodium dihydrogen phosphate solution (50 mL. times.1) and saturated brine (50 mL. times.1) in this order. After the organic layer was dried and concentrated, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1-1/1) to give compound 25 as a pale yellow oil (280mg, yield 34.14%).

¹H NMR(400MHz,CDCl₃)δ5.28–5.11(m,3H),5.06–4.73(m,1H),4.68-4.58(m,1H),4.15–3.97(m,5H),3.88-3.82(m,1H),3.77(d,3H),3.55-3.43(m,4H),2.41–2.34(m,2H),2.32-2.26(m,2H),2.20(dd,3H),1.80(s,3H),1.29–1.17(m,9H),1.09(dd,3H).

³¹P NMR(162MHz,CDCl₃)δ27.27,26.30.

LC-MS(m/z):570.2[M+1]。

Example 26:

(E) -6- [4- [ [ [ (1S) -2-hexyloxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (compound 26)

(E)-6-[4-[[[(1S)-2-hexoxy-1-methyl-2-oxo-ethyl]amino]-(methoxymethyl)phosphoryl]oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoic acid

The first step is as follows: benzyl (E) -6- (4-hydroxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl) -4-methyl-hex-4-enoate (26B)

benzyl(E)-6-(4-hydroxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl)-4-methyl-hex-4-enoate

Dissolving 26A (4g,12.49mmol) in dichloromethane (50mL), adding 1-2 drops of N, N-dimethylformamide and thionyl chloride (1.78g,14.96mmol) in sequence, stirring at room temperature for 1h, removing the solvent residue under reduced pressure, and dissolving in 10mL dichloromethane to obtain an acyl chloride solution. In another three-necked flask, benzyl alcohol (2g,18.49mmol) and pyridine (2.92g,37.44mmol) were dissolved in dichloromethane (30mL), cooled to 0 deg.C, the acid chloride solution was added slowly, and stirring was continued for 1h while warming to room temperature. The reaction solution was washed with dilute hydrochloric acid (1M, 50 mL. times.2) and saturated brine (50 mL. times.1) in this order. The organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 20/1-5/1) to give compound 26B as a pale yellow oil (3g, 58.52% yield).

¹H NMR(400MHz,CDCl₃)δ7.67(s,1H),7.34–7.28(m,5H),5.28-5.22(m,1H),5.17(s,2H),5.05(d,2H),3.79–3.68(m,3H),3.37(d,2H),2.51–2.39(m,2H),2.39–2.27(m,2H),2.13(s,3H),1.79(s,3H).

LC-MS(m/z):411.2[M+1]。

The second step is that: n-hexyl (2S) -2- (tert-butoxycarbonylamino) propionate (26C)

hexyl(2S)-2-(tert-butoxycarbonylamino)propanoate

26A (3g,15.86mmol) was dissolved in dichloromethane (20mL), n-hexanol (2.43g,23.78mmol),1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (4.56g,23.79mmol) and 4-dimethylaminopyridine (2.9g,23.78mmol) were added, the reaction was reacted at room temperature for 4 hours, the reaction solution was washed with a saturated aqueous solution of sodium dihydrogenphosphate (20mL), separated, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue as the title compound 26C, a colorless liquid (3.5g, yield 80.73%).

LC-MS(m/z):296.2[M+23]。

The third step: n-hexyl (2S) -2-aminopropionate hydrochloride (26D)

hexyl(2S)-2-aminopropanoate hydrochloride

26C (3.5g,12.80mmol) was dissolved in ethyl acetate (20mL), dried HCl gas was bubbled through at room temperature for 4h, the solvent was removed under reduced pressure, the residue was slurried with ethyl acetate/petroleum ether (1/1(v/v),20mL) and filtered to give compound 26D as a white solid (2.5g, 93.13% yield).

¹H NMR(400MHz,CDCl₃)δ8.74(s,3H),4.34–4.05(m,3H),1.84–1.52(m,5H),1.46–1.19(m,6H),0.89(t,3H).

The fourth step: benzyl (E) -6- [4- [ [ [ (1S) -2-hexyloxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxy-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoate (26E)

benzyl(E)-6-[4-[[[(1S)-2-hexoxy-1-methyl-2-oxo-ethyl]amino]-(methoxymethyl)phosphoryl]oxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate。

Compound 3B (1g,2.44mmol) was dissolved in dichloromethane (30mL), and (methoxymethyl) phosphonodichloride (0.40g,2.45mmol) was added under ice-cooling, and triethylamine (0.8g,7.91mmol) was slowly added, followed by stirring at 0 ℃ for 20min and addition of 3D (0.53g,2.46 mmol). After warming to room temperature and stirring for further 30min, dichloromethane (50mL) was added, and the mixture was washed with saturated sodium dihydrogen phosphate solution (50 mL. times.1) and saturated brine (50 mL. times.1) in this order. After the organic layer was dried and concentrated, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1 to 1/1) to give compound 26E as a pale yellow oil (800mg, yield 48.70%).

LC-MS(m/z):674.1[M+1]。

The fifth step: (E) -6- [4- [ [ [ (1S) -2-hexyloxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxy-6-methoxy-7-methyl-3-oxo-1 hydro-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (compound 26)

Compound 26E (0.8g,1.19mmol) was dissolved in methylene chloride/ethanol (1/1,20mL), and triethylamine (24mg,0.24mmol), palladium acetate (27mg,0.12mmol) and triethylsilane (415mg,3.57mmol) were added in this order, followed by stirring at room temperature for 6 h. The mixture was washed with saturated ammonium chloride solution (10mL x 1) and saturated brine (10mL x 1) in that order. The organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1-1/1) to give compound 26 as a pale yellow oil (400mg, yield 57.60%).

¹H NMR(400MHz,CDCl₃)δ5.28–5.11(m,3H),4.79-4.67(m,1H),4.13–3.86(m,5H),3.84–3.70(m,4H),3.59–3.41(m,4H),2.41(dd,2H),2.29(t,2H),2.21(t,3H),1.76(d,3H),1.67–1.58(m,1H),1.49(dd,1H),1.39(d,1H),1.36–1.17(m,9H),0.90-0.85(m,3H).

³¹P NMR(162MHz,CDCl₃)δ27.28,26.36.

LC-MS(m/z):584.2[M+1]。

Example 27:

methyl (E) -6- [4-bis [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid ester (Compound 27)

methyl(E)-6-[4-bis[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate

Compound 24B (0.5g,1.50mmol) is dissolved in dichloromethane (20mL), cooled to-78 deg.C, phosphorus oxychloride (0.25g,1.53mmol) is added, and triethylamine (0.16g,1.58mmol) is added dropwise. After stirring at this temperature for 20 minutes, L-alanine isopropyl ester hydrochloride (0.5g,2.98mmol) and triethylamine (0.64g,6.32mmol) were added, and the mixture was warmed to room temperature and stirred for 30 minutes. Dichloromethane (50mL) was added and the mixture was washed successively with saturated sodium dihydrogen phosphate solution (50 mL. times.1) and saturated brine (50 mL. times.1). After the organic layer was dried and concentrated, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1 to 1/1) to give compound 27 as a pale yellow oil (0.25g, yield 26.01%).

¹H NMR(400MHz,CDCl₃)δ5.26-5.23(m,1H),5.15(s,2H),5.06-4.92(m,2H),4.71(t,1H),4.20(t,1H),4.06-3.94(m,2H),3.76(s,3H),3.63–3.55(m,5H),2.44–2.35(m,2H),2.30(dd,2H),2.18(s,3H),1.79(s,3H),1.41(d,3H),1.37(d,3H),1.26(d,6H),1.20(dd,6H).

³¹P NMR(162MHz,CDCl₃)δ11.81.

LC-MS(m/z):641.2[M+1]。

Example 28:

ethyl (E) -6- [4-bis [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 hydroiso-benzofuran-5-yl ] -4-methyl-hex-4-enoic acid ester (Compound 28)

ethyl(E)-6-[4-bis[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate。

Compound 24B (0.5g,1.44mmol) is dissolved in dichloromethane (20mL), cooled to-78 deg.C, phosphorus oxychloride (0.24g,1.47mmol) is added, and triethylamine (0.15g,1.48mmol) is added dropwise. After stirring at this temperature for 20min, L-alanine isopropyl hydrochloride (0.50g,2.98mmol) was added, triethylamine (0.6g,5.92mmol) was added, and stirring was continued for 30min at room temperature. Dichloromethane (50mL) was added and the mixture was washed successively with saturated sodium dihydrogen phosphate solution (50 mL. times.1) and saturated brine (50 mL. times.1). After the organic layer was dried and concentrated, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1 to 1/1) to give compound 28 as a pale yellow oil (0.25mg, yield 26.53%).

¹H NMR(400MHz,CDCl₃)δ5.24(d,1H),5.15(s,2H),5.08–4.99(m,1H),4.99–4.88(m,1H),4.72(s,1H),4.20(s,1H),4.11–3.90(m,4H),3.76(s,3H),3.58(d,2H),2.41–2.33(m,2H),2.30(d,2H),2.18(d,3H),1.79(s,3H),1.39(dd,6H),1.29–1.15(m,15H).

³¹P NMR(162MHz,CDCl₃)δ10.17.

LC-MS(m/z):655.3[M+1].

Example 29:

(E) -6- [4-bis [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (Compound 29)

(E)-6-[4-bis[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoic acid。

The first step is as follows: benzyl (E) -6- [4-bis [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoate (29A)

benzyl(E)-6-[4-bis[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate。

Compound 26B (2.5g,6.09mmol) was dissolved in dichloromethane (100mL), cooled to-78 deg.C, phosphorus oxychloride (0.93g,6.08mmol) was added, and triethylamine (0.65g,6.43mmol) was added dropwise. After stirring at this temperature for 20 minutes, L-alanine isopropyl ester hydrochloride (2.05g,12.39mmol) and triethylamine (2.60g,25.72mmol) were added, and the mixture was warmed to room temperature and stirred for 30 minutes. Dichloromethane (100mL) was added and washed successively with saturated sodium dihydrogen phosphate solution (100mL × 1) and saturated brine (100mL × 1). After the organic layer was dried and concentrated, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1 to 1/1) to give compound 29A as a pale yellow oil (3.1g, yield 71.00%).

LC-MS(m/z):717.3[M+1]。

The second step is that: (E) -6- [4-bis [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (Compound 29)

Compound 29A (3.2g,4.46mmol) was dissolved in methylene chloride/ethanol (1/1,50mL), triethylamine (45mg,0.45mmol), palladium acetate (101mg,0.45mol) and triethylsilane (1.56g,13.41mmol) were added in this order, and the mixture was stirred at room temperature for 12 h. The reaction mixture was washed with a saturated solution of ammonium chloride (20 mL. times.1) and saturated brine (20 mL. times.1) in this order. The organic layer was dried and concentrated, and the residue was slurried with ethyl acetate/petroleum ether (1/1,50mL) and filtered to give compound 29 as a white solid (2g, 71.56% yield).

¹H NMR(400MHz,CDCl₃)δ5.26(t,1H),5.14(s,2H),5.05–4.96(m,2H),4.83(s,1H),4.08(dd,1H),3.97(dd,1H),3.76(s,3H),3.53(d,2H),2.51–2.38(m,2H),2.32(dd,2H),2.18(d,3H),1.79(d,3H),1.41(dd,6H),1.29–1.16(m,12H).

³¹P NMR(162MHz,CDCl₃)δ12.51.

LC-MS m/z＝627.3[M+1]。

Example 30:

(E) -6- [4-bis [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (compound 30)

(E)-6-[4-bis[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoic acid

The first step is as follows: benzyl ((E) -6- [4-bis [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl ] -4-methyl-hex-4-enoate (30A)

benzyl(E)-6-[4-bis[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate.

Compound 3B (1.7g,4.14mmol) was dissolved in dichloromethane (50mL), cooled to-78 deg.C, phosphorus oxychloride (0.63g,4.12mmol) was added, and triethylamine (0.45g,4.45mmol) was added dropwise. After stirring at this temperature for 20 minutes, L-alanine ethyl ester hydrochloride (1.27g,8.27mmol) and triethylamine (1.8g,17.80mmol) were added and stirring was continued at room temperature for 30 minutes. Dichloromethane (50mL) was added and the mixture was washed successively with saturated sodium dihydrogen phosphate solution (50 mL. times.1) and saturated brine (50 mL. times.1). The organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1-1/1) to give compound 30A as a pale yellow oil (1.9g, yield 66.64%).

LC-MS(m/z):689.3[M+1]。

The second step is that: ((E) -6- [4-bis [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (Compound 30)

(E)-6-[4-bis[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoic acid。

Compound 30A (1.9g,2.76mmol) was dissolved in methylene chloride/ethanol (1/1,50mL), triethylamine (28mg,0.28mmol), palladium acetate (62mg,0.28mmol) and triethylsilane (0.96g,8.26mmol) were added in that order, and the mixture was stirred at room temperature for 12 h. The reaction mixture was washed with a saturated solution of ammonium chloride (20 mL. times.1) and saturated brine (20 mL. times.1) in this order. The organic layer was dried and concentrated, and the residue was slurried with ethyl acetate/petroleum ether (1/1,50mL) and filtered to give compound 30 as a white solid (1g, 60.53% yield).

¹H NMR(400MHz,CDCl₃)δ5.25(d,1H),5.15(s,2H),4.95(s,1H),4.24–4.05(m,5H),4.02(dd,1H),3.77(s,3H),3.52(s,2H),2.44-2.41(m,2H),2.37–2.25(m,2H),2.19(s,3H),1.79(d,3H),1.43(dd,6H),1.32–1.20(m,6H).

³¹P NMR(162MHz,CDCl₃)δ10.96.

LC-MS(m/z):599.2[M+1].

Example 31:

(E) -6- [4-bis [ (2-ethoxy-2-oxo-ethyl) amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (compound 31)

(E)-6-[4-bis[(2-ethoxy-2-oxo-ethyl)amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoic acid。

The first step is as follows: benzyl (E) -6- [4-bis [ (2-ethoxy-2-oxo-ethyl) amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (31A)

benzyl(E)-6-[4-bis[(2-ethoxy-2-oxo-ethyl)amino]phosphoryloxy-6-methoxy-7-methyl-3-oxo-1H-isobenzofuran-5-yl]-4-methyl-hex-4-enoate。

Compound 26B (1.5g,3.65mmol) is dissolved in dichloromethane (50mL), cooled to-78 deg.C, phosphorus oxychloride (0.56g,3.66mmol) is added, and triethylamine (0.4g,3.95mmol) is added dropwise. After stirring at this temperature for 20 minutes, glycine ethyl ester hydrochloride (1.02g,7.31mmol) was added, triethylamine (1.6g,15.80mmol) was added, and stirring was continued for 30min at room temperature. Dichloromethane (50mL) was added and the mixture was washed successively with saturated sodium dihydrogen phosphate solution (50 mL. times.1) and saturated brine (50 mL. times.1). The organic layer was dried and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1-1/1) to give compound 31A as a pale yellow oil (1.5g, yield 62.21%).

LC-MS(m/z):661.3[M+1],683.3[M+23]。

The second step is that: (E) -6- [4-bis [ (2-ethoxy-2-oxo-ethyl) amino ] phosphoryl-6-methoxy-7-methyl-3-oxo-1 h-isobenzofuran-5-yl ] -4-methyl-hex-4-enoic acid (compound 31)

Compound 8A (1.5g,2.27mmol) was dissolved in methylene chloride/ethanol (1/1(v/v),40mL), triethylamine (0.23g,0.23mmol), palladium acetate (51mg,0.23mmol) and triethylsilane (792mg,6.81mmol) were added in that order, and the mixture was stirred at room temperature for 12 h. The mixture was washed with saturated ammonium chloride solution (20mL x 1) and saturated brine (20mL x 1) in that order. The organic layer was dried and concentrated, and purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 5/1-1/1) to give compound 31 as a white solid (0.7g, 54.05% yield).

¹H NMR(400MHz,CDCl₃)δ5.33–5.22(m,1H),5.15(s,2H),4.25–4.12(m,4H),3.93–3.72(m,7H),3.59–3.43(m,2H),2.42(t,2H),2.31(t,2H),2.20(d,3H),1.76(t,3H),1.32–1.21(m,6H).

³¹P NMR(162MHz,CDCl₃)δ12.91.

LC-MS(m/z):571.2[M+1]。

Example 32

Ethyl (2S) -2- [ [ [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] - [ [4- (4-hydroxy-3, 5-diisopropyl-phenyl) -2, 6-diisopropyl-phenoxy ] methoxy ] phosphoryl ] amino ] propionate (compound 32)

ethyl(2S)-2-[[[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]-[[4-(4-hydroxy-3,5-diisopropyl-phenyl)-2,6-diisopropyl-phenoxy]methoxy]phosphoryl]amino]propanoate

The first step is as follows:

4- [4- (chloromethoxy) -3, 5-diisopropyl-phenyl ] -2, 6-diisopropyl-phenol (32B)

4-[4-(chloromethoxy)-3,5-diisopropyl-phenyl]-2,6-diisopropyl-phenol

Compound 32A (2g,5.65mmol) was dissolved in tetrahydrofuran (20mL), sodium hydride (0.13g,5.33mmol) was added slowly at 0 deg.C, and stirring was continued for 20min while naturally warming to room temperature. Bromochloromethane (2.20g,17.00mmol) was added, the temperature was raised to 50 ℃ and the reaction was stirred for 4 h. The solvent was removed under reduced pressure, extracted with ethyl acetate (20mL), and washed with brine (20 mL. times.1). After the organic layer was concentrated, the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)400/1 to 100/1) to obtain compound 32B as a yellow oil (500mg, yield 21.96%).

¹H NMR(400MHz,DMSO-d₆)δ8.14(s,1H),7.25(s,2H),7.13(s,2H),5.94(s,2H),3.40–3.33(m,4H),1.26–1.18(m,24H).

The second step is that:

ethyl (2S) -2- [ [ benzyloxy- [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] phosphoryl ] amino ] propionate (32D)

ethyl(2S)-2-[[benzyloxy-[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate

Benzyl alcohol (5g,46.24mmol) was dissolved in dichloromethane (250mL) and, under nitrogen, phosphorus oxychloride (7.1g,46.41mmol) was added at-78 deg.C, triethylamine (4.70g,46.47mmol) was slowly added dropwise at 78 deg.C and stirring was continued for 20 min. L-alanine ethyl ester hydrochloride (14.28g,92.96mmol) was added, triethylamine (18.8g,185.88mmol) was slowly added dropwise at 78 deg.C, and the mixture was warmed to room temperature and stirred for 30 minutes. The reaction mixture was washed successively with saturated solution of disodium hydrogenphosphate (200 mL. times.1) and saturated solution of sodium chloride (200 mL. times.1). The organic layer was dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)5/1 to 1/1) to give compound 32D as a pale yellow oil (10g, yield: 55.97%).

¹H NMR(400MHz,CDCl₃)δ7.43–7.25(m,5H),5.01(d,2H),4.25–4.04(m,4H),4.04–3.82(m,2H),3.28(t,2H),1.43–1.29(m,6H),1.28–1.15(m,6H).

³¹P NMR(162MHz,CDCl₃)δ13.49

LC-MS m/z＝387.3[M+1]。

The third step:

bis [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] phosphinic acid (32E)

bis[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]phosphinic acid

Compound 32D (1g,2.58mmol) was dissolved in methanol (20mL), triethylamine (258mg,2.55mmol) and palladium on carbon (1g,10 wt.%) were added sequentially, stirred under a hydrogen balloon for 2h, filtered, and the filtrate was concentrated to give compound 32E as a pale yellow oil (1.1g) which was used directly in the next reaction.

LC-MS m/z＝297.1[M+1]

The fourth step:

Compound 32B (500mg,1.24mmol) and compound 32E (550mg,1.86mmol) were dissolved in N, N-dimethylformamide (10mL), potassium carbonate (256mg,1.86mmol) was added, and the mixture was stirred at 50 ℃ for 4 hours. Ethyl acetate (50mL) was added for extraction and the mixture was washed with saturated brine (50 mL. times.5). The organic layer was dried with anhydrous sulfuric acid and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)5/1 to 1/1) to obtain compound 32 as a yellow oil (0.35g, 42.59% yield).

¹H NMR(400MHz,CDCl₃)δ7.25–7.21(m,2H),7.18(s,2H),5.52–5.36(m,2H),4.20–4.08(m,4H),3.96(dd,2H),3.45–3.36(m,2H),3.22(dd,2H),1.39–1.22(m,36H).

³¹P NMR(162MHz,CDCl₃)δ12.19.

LC-MS m/z＝661.4[M-1]

Example 33

(2S) -ethyl 2- [ [ [4- (4-hydroxy-3, 5-diisopropylphenyl) -2, 6-diisopropyl-phenoxy ] methoxy- (methoxymethyl) phosphoryl ] amino ] propionate (compound 33)

ethyl(2S)-2-[[[4-(4-hydroxy-3,5-diisopropyl-phenyl)-2,6-diisopropyl-phenoxy]methoxy-(methoxymethyl)phosphoryl]amino]propanoate

Compound 32B (1.8g,4.4mmol) and N- [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] - (methoxymethyl) phosphoramidic acid (1.0g,4.4mmol) were dissolved in N, N-dimethylformamide (20mL), potassium carbonate (1.2g,8.8mmol) was added, and the mixture was stirred at 50 ℃ for 4 hours. Ethyl acetate (50mL) was added for extraction and the mixture was washed with saturated brine (50 mL. times.5). The organic layer was dried with anhydrous sulfuric acid and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate (v/v)5/1 to 1/1) to obtain compound 33 as a yellow oil (0.5g, yield 19.2%).

¹H NMR(400MHz,CDCl₃)δ7.22(s,2H),7.18(d,2H),5.62–5.52(m,1H),5.54–5.43(m,1H),4.26–4.02(m,3H),3.76-3.72(m,2H),3.46–3.16(m,8H),1.41–1.30(m,15H),1.29–1.24(m,15H).

31P NMR(162MHz,CDCl3)δ25.88,25.08。

Example 34

Compound 3: ethyl (2S) -2- [ [ [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] - [4- (4-hydroxy-3, 5-diisopropyl-phenyl) -2, 6-diisopropyl-phenoxy ] phosphoryl ] amino ] propanoate (compound 34)

ethyl(2S)-2-[[[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]-[4-(4-hydroxy-3,5-diisopropyl-phenyl)-2,6-diisopropyl-phenoxy]phosphoryl]amino]propanoate

Dissolving phosphorus oxychloride (1.54g, 10mmol) in 100ml dichloromethane, cooling to-30 ℃ under the protection of nitrogen, dropwise adding a mixed dichloromethane solution of 4- (4-hydroxy-3, 5-diisopropylphenyl) -2, 6-diisopropylphenol (3.56g, 10mmol) and triethylamine (2.03g, 20mmol) in 30ml, continuing stirring for 30min after dropwise adding, adding L-alanine ethyl ester hydrochloride (3.86g, 25.1mmol), naturally raising the temperature to room temperature and reacting for 2 h. After the reaction was completed, the reaction mixture was washed once with 150ml of a saturated aqueous solution of sodium dihydrogenphosphate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography, ethyl acetate/petroleum ether (v/v) ═ 0: 1-1: 1 as eluent, compound 34 was obtained as a colorless oil (1.0g, 16% yield).

1H NMR(400MHz,CDCl3)δ7.22(s,2H),7.17(s,2H),4.15(dddd,6H),3.61–3.44(m,3H),3.36(t,1H),3.28–3.16(m,2H),1.43(d,3H),1.36–1.23(m,33H).

³¹P NMR(162MHz,CDCl₃)δ8.34

LC-MS m/z＝633.4[M+1]

Examples 35 and 36

(2S) -ethyl 2- [ [ [2- [ (1R) -1-cyclopropylethyl ] -4- [3- [ (1R) -1-cyclopropylethyl ] -4-hydroxy-5-isopropyl-phenyl ] -6-isopropyl-phenoxy ] - (methoxymethyl) phosphoryl ] amino ] alanine (Compound 35)

ethyl(2S)-2-[[[2-[(1R)-1-cyclopropylethyl]-4-[3-[(1R)-1-cyclopropylethyl]-4-hydroxy-5-isopropyl-phenyl]-6-isopropyl-phenoxy]-(methoxymethyl)phosphoryl]amino]propanoate

(2S) -ethyl 2- [ [ [2- [ (1R) -1-cyclopropylethyl ] -4- [3- [ (1R) -1-cyclopropylethyl ] -4- [ [ [ (1S) -2-ethoxy-1-methyl-2-oxo-ethyl ] amino ] - (methoxymethyl) phosphoryl ] oxo-5-isopropyl-phenoxy ] -6-isopropyl-phenoxy ] - (methoxymethyl) phosphoryl ] amino ] alanine (Compound 36)

Ethyl(2S)-2-[[[2-[(1R)-1-cyclopropylethyl]-4-[3-[(1R)-1-cyclopropylethyl]-4-[[[(1S)-2-ethoxy-1-methyl-2-oxo-ethyl]amino]-(methoxymethyl)phosphoryl]oxy-5-isopropyl-phenyl]-6-isopropyl-phenoxy]-(methoxymethyl)phosphoryl]amino]propanoate

Diphosphoryl chloride (1.80g, 11.05mmol) was dissolved in 20mL dry dichloromethane and reacted at-30 ℃ with triethylamine (3.00g, 29.65mmol) and a solution of L-alanine ethyl ester (1.70g, 11.07mmol) in dichloromethane (10mL) under nitrogen, for 30min after dropping, 35A (3.00g, 7.38mmol) was added and reacted at room temperature for 2 h. Washing with 20mL of saturated aqueous sodium dihydrogen phosphate solution once, washing with 20mL of saturated brine, drying over anhydrous sodium sulfate, concentrating under reduced pressure, and column chromatography using ethyl acetate/petroleum ether (v/v) ═ 1/1-1/0 as an eluent, yielded compound 35, a yellow solid (0.80g, yield 17.67%) and compound 36, a yellow solid (0.5g, yield 11.80%).

Compound 35:

LC-MS m/z＝612.4[M-1].

¹H NMR(400MHz,CDCl₃)δ7.40-7.37(dd,1H),7.28-7.25(m,2H),7.22-7.20(dd,1H),5.09(s,1H),4.14-4.06(m,3H),3.89-3.76(m,2H),3.58-3.51(m,1H),3.47(dd,3H),3.25-3.16(m,1H),2.82-2.76(m,1H),2.57-2.47(m,1H),1.38-1.24(m,22H),1.20(t,3H),1.16-1.08(m,1H),1.02-0.95(m,1H),0.59-0.48(m,3H),0.42-0.35(m,1H),0.32-0.21(m,4H).

compound 36:

LC-MS m/z＝843.5[M+23].

¹H NMR(400MHz,d₆-DMSO)δ7.40-7.39(m,2H),7.26-7.25(m,2H),5.54-5.43(m,2H),4.13-4.01(m,4H),3.92-3.78(m,6H),3.59-3.50(m,2H),3.41-3.39(m,6H),2.79-2.72(m,2H),1.19-1.13(m,30H),1.06-1.04(m,2H),0.51-0.50(m,2H),0.30-0.19(m,6H).

example 37

Isopropyl (2S) -2- [ [ [4- [3- [ (1R) -1-cyclopropylethyl ] -4-hydroxy-5-isopropyl-phenyl ] -2, 6-diisopropyl-phenoxy ] - (methoxymethyl) phosphoryl ] amino ] propionate (compound 37)

Isopropyl(2S)-2-[[[4-[3-[(1R)-1-cyclopropylethyl]-4-hydroxy-5-isopropyl-phenyl]-2,6-diisopropyl-phenoxy]-(methoxymethyl)phosphoryl]amino]propanoate

(methoxymethyl) phosphonyl dichloride (0.64g,3.94mmol) is dissolved in 10mL of dry dichloromethane, a mixture of triethylamine (1.60g, 15.76mmol) and L-isopropyl alanine (0.66g, 3.94mmol) is added dropwise at-30 ℃ under the protection of nitrogen, reaction is carried out for 30min after dropwise addition, 37A (1.00g, 2.63mmol) is added, and reaction is carried out for 5h at room temperature. Washing with 20mL of saturated aqueous sodium dihydrogen phosphate solution and 20mL of saturated brine, drying over anhydrous sodium sulfate, concentrating under reduced pressure, performing column chromatography, eluting with ethyl acetate/petroleum ether (v/v) ═ 1/1-1/0 to give compound 37 as a yellow solid (0.2g, yield 13.0%)

LC-MS m/z＝600.5[M-1].

¹H NMR(400MHz,CDCl₃)δ7.25-7.18(m,4H),5.05-4.93(m,2H),4.15-4.09(m,1H),3.93-3.86(m,2H),3.57-3.48(m,5H),3.23-3.18(m,1H),2.56-2.53(m,1H),1.39-1.17(m,30H),1.15-1.09(m,1H),0.63-0.50(m,2H),0.29-0.21(m,2H).

Example 38

(2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS) -6,12 b-bis (acetoxy) -12- (benzoyloxy) -2a,3,4,4a,5,6,9,10,11,12,12a,12 b-dodecahydro-4, 11-dihydroxy-4 a,8,13, 13-tetramethyl-5-oxo-7, 11-methano 1 hydro-cyclodecan [3,4] benzo [1,2-b ] oxetan-9-yl (α R, β S) - β - (benzoylamino) - α -isopropyl (2S) -2- [ [ methoxymethyl- [4- [ oxycarbonyloxymethyl ] phenoxy ] phosphoryl ] amino ] propionate-phenylpropionate (Compound 38).

(2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-6,12b-bis(acetyloxy)-12-(benzoyloxy)-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-4,11-dihydroxy-4a,8,13,13-tetramethyl-5-oxo-7,11-methano-1H-cyclodeca[3,4]benz[1,2-b]oxet-9-yl(αR,βS)-β-(benzoylamino)-α-isopropyl(2S)-2-[[methoxymethyl-[4-[oxycarbonyloxymethyl]phenoxy]phosphoryl]amino]propanoate-Benzenepropionate

Intermediate 2(0.202g, 0.586mmol) was dissolved in tetrahydrofuran (5ml), cooled to 0 ℃ under nitrogen, triphosgene (0.087g, 0.293mmol) was added, diisopropylethylamine (0.114g, 0.878mmol) was added and stirring continued for 30min after addition was complete. Paclitaxel (38A) (0.500g, 0.586mmol) was dissolved in dry dichloromethane (5ml), 4-dimethylaminopyridine (0.107g, 0.879mmol) was added, and the reaction mixture was slowly added in ice bath, and then naturally warmed to room temperature for reaction for 2 hours. Dichloromethane (20ml) and saturated sodium dihydrogen phosphate (30ml) were added, the mixture was separated, the organic layer was washed with saturated sodium chloride (20ml), dried over anhydrous sodium sulfate, and the crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) ═ 4: 1to 1:4) to give compound 38 as a white solid (0.200g, yield 27.9%).

1H NMR(400MHz,DMSO-d6)δ9.27(d,1H),8.03–7.96(m,2H),7.87–7.81(m,2H),7.73(t,1H),7.68-7.62(m,2H),7.58-7.54(m,1H),7.50-7.42(m,6H),7.38-7.34(m,2H),7.22-7.14(m,3H),6.32(s,1H),5.84(t,1H),5.72-5.62(m,1H),5.56(t,1H),5.44(d,1H),5.38(d,1H),5.16(s,2H),4.94-4.88(m,2H),4.86-4.80(m,1H),4.64(s,1H),4.18-4.10(m,1H),4.08–3.97(m,2H),3.93–3.82(m,1H),3.80-3.72(m,2H),3.61(d,1H),3.37(dd,3H),2.40–2.29(m,1H),2.27(s,3H),2.12(s,3H),1.89–1.78(m,4H),1.66(t,1H),1.60–1.47(m,4H),1.20-1.12(m,9H),1.03(d,6H).

³¹P NMR(162MHz,DMSO-d6)δ24.99,24.15.

Example 39

(2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS) -6,12 b-bis (acetoxy) -12- (benzoyloxy) -2a,3,4,4a,5,6,9,10,11,12,12a,12 b-dodecahydro-11-hydroxy-4 a,8,13, 13-tetramethyl-5-oxo-7, 11-methano-1-hydro-cyclodecano [3,4] benzo [1,2-b ] oxetan-9-yl (. alpha.R,. beta.S) -beta- (benzoylamino) -alpha, 4-bis [ isopropyl (2S) -2- [ [ methoxymethyl- [4- [ oxycarbonyloxymethyl ] phenoxy ] phosphoryl ] amino ] propanoate ] -phenylpropionate (compound 39).

(2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-6,12b-bis(acetyloxy)-12-(benzoyloxy)-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-11-hydroxy-4a,8,13,13-tetramethyl-5-oxo-7,11-methano-1H-cyclodeca[3,4]benz[1,2-b]oxet-9-yl(αR,βS)-β-(benzoylamino)-α,4-bis[isopropyl(2S)-2-[[methoxymethyl-[4-[oxycarbonyloxymethyl]phenoxy]phosphoryl]amino]propanoate]-Benzenepropionate

Intermediate 2(0.505g, 1.46mmol) was dissolved in tetrahydrofuran (10ml), cooled to 0 ℃ under nitrogen, triphosgene (0.217g, 0.732mmol) was added, diisopropylethylamine (0.227g, 1.76mmol) was added and stirring was continued for 30 minutes after the addition was complete. Paclitaxel (0.500g, 0.586mmol) was dissolved in dry dichloromethane (5ml), 4-dimethylaminopyridine (0.107g, 0.879mmol) was added, and the reaction mixture was slowly added in ice bath, and then naturally warmed to room temperature to react for 2 hours. Dichloromethane (20ml) and saturated sodium dihydrogen phosphate (30ml) were added, the mixture was separated, the organic layer was washed with saturated sodium chloride solution (20ml), dried over anhydrous sodium sulfate, and the crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (petroleum ether: ethyl acetate (v/v) ═ 4: 1to 1:4) to give compound 39 as a white solid (0.300g, yield 32.1%).

¹H NMR(400MHz,DMSO-d6)δ9.30(d,1H),8.02–7.95(m,2H),7.84(d,2H),7.75(t,1H),7.67(t,2H),7.57(t,1H),7.53–7.42(m,6H),7.42–7.34(m,4H),7.24–7.14(m,5H),6.27(s,1H),5.84(t,1H),5.74-5.62(m,2H),5.51(t,1H),5.46–5.34(m,3H),5.21(s,2H),5.13–5.02(m,2H),4.98(d,1H),4.91–4.78(m,2H),4.73(s,1H),4.10-4.00(m,2H),3.92–3.81(m,2H),3.81–3.67(m,5H),3.42–3.32(m,6H),2.50-2.42(m,1H),2.28(s,3H),2.14(s,3H),1.83-1.70(m,5H),1.63(s,3H),1.56–1.45(m,1H),1.21–1.09(m,18H),1.02(d,6H).

³¹P NMR(162MHz,DMSO-d6)δ24.95,24.77,24.12,23.95.

LC-MS(M/Z)＝799.0[M/2+1]

Example 40

(2AR,4S,4AS,6R,9S,11S,12S,12AR,12bS) -6,12b bis (acetoxy) -12- (benzoyloxy) -2a,3,4,4a,5,6,9,10,11,12,12a,12 b-dodeca-4, 11-dihydroxy-4 a,8,13, 13-tetramethyl-5-oxo-7, 11-methanol-1H-cyclodeca [3,4] benzo [1,2-b ] oxetan-9-yl (alpha R, beta S) -beta- (benzoylamino) -alpha-isopropyl (2S) -2- [ [ [4- (carboxyoxymethyl) phenoxy ] - [ [ (1S) -2-isopropyl-1-methyl-2-oxo-ethyl ] amino ] phosphoryl Yl ] amino ] propionate-phenylpropionate (Compound 40)

(2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-6,12b-bis(acetyloxy)-12-(benzoyloxy)-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-4,11-dihydroxy-4a,8,13,13-tetramethyl-5-oxo-7,11-methano-1H-cyclodeca[3,4]benz[1,2-b]oxet-9-yl(αR,βS)-β-(benzoylamino)-α-isopropyl(2S)-2-[[[4-(carboxyoxymethyl)phenoxy]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate-Benzenepropionate

Dissolving the intermediate 1(0.43g,1mmol) in 5ml tetrahydrofuran, adding triphosgene (0.148g, 0.5mmol) at 0 ℃ under the protection of nitrogen, adding N, N-diisopropylethylamine (0.194g, 1.5mmol) dropwise, and reacting for 30min after dropwise addition. To obtain reaction solution 1, paclitaxel (38A) (0.854g, 1mmol) was dissolved in 20ml dichloromethane, N-diisopropylethylamine (0.323g, 2.5mmol) was added, the temperature was reduced to 0 ℃, reaction solution 1 was dropped, and the reaction was continued for 2 hours. Washing the precipitate once with 30ml of saturated sodium dihydrogen phosphate aqueous solution, drying the precipitate with anhydrous sodium sulfate, concentrating the precipitate under reduced pressure, and carrying out column chromatography by using ethyl acetate/petroleum ether (v/v) 0/1-1/1 as an eluent. Compound 40 was obtained as a white solid (0.15g, 11% yield).

¹H NMR(400MHz,DMSO-d6)δ9.26(d,1H),8.06–7.95(m,2H),7.89–7.79(m,2H),7.73(t,1H),7.64(t,2H),7.60–7.53(m,1H),7.53–7.38(m,6H),7.34(d,2H),7.27–7.10(m,3H),6.31(s,1H),5.83(t,1H),5.54(t,1H),5.43(d,1H),5.36(d,1H),5.24-5.16(m,4H),4.95–4.79(m,4H),4.64(s,1H),4.16-4.10(m,1H),4.07–3.94(m,2H),3.86-3.77(m,2H),3.60(d,1H),2.41–2.29(m,1H),2.27(s,3H),2.12(s,3H),1.99(s,2H),1.81(s,3H),1.65(t,1H),1.51(s,3H),1.25(dd,6H),1.18–1.14(m,12H),1.02(d,6H).

³¹P NMR(162MHz,DMSO-d6)δ10.74.

EXAMPLE 41

(2AR,4S,4AS,6R,9S,11S,12S,12AR,12bS) -12b- (acetoxy) -12- (benzoyloxy) -2a,3,4,4a,5,6,9,10,11,12,12a,12 b-dodecahydro-4, 6, 11-trihydroxy-4 a,8,13, 13-tetramethyl-5-oxo-7, 11-methanol-1H-cyclododecene [3,4] benzo [1,2-b ] oxetan-9-yl (α R, β S) - β - [ [ ((1, 1-dimethylethoxy) carbonyl ] amino ] - α -isopropyl (2S) -2- [ [ [4- (carboxyoxymethyl) phenoxy ] [ (1S) -2-isopropoxy-1-methyl-) 2-oxo-ethyl ] amino ] phosphoryl ] amino ] propionate-phenylpropionate (Compound 41)

(2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-12b-(acetyloxy)-12-(benzoyloxy)-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-4,6,11-trihydroxy-4a,8,13,13-tetramethyl-5-oxo-7,11-methano-1H-cyclodeca[3,4]benz[1,2-b]oxet-9-yl(αR,βS)-β-[[(1,1-dimethylethoxy)carbonyl]amino]-α-isopropyl(2S)-2-[[[4-(carboxyoxymethyl)phenoxy]-[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphoryl]amino]propanoate-Benzenepropionate

Intermediate 1(0.43g,1mmol) was dissolved in 5ml tetrahydrofuran, triphosgene (0.148g, 0.5mmol) was added at 0 ℃ under nitrogen protection, and N, N-diisopropylethylamine (0.194g, 1.5mmol) was added dropwise and reacted for 30min after the addition. To obtain reaction solution 1, docetaxel (41A) (0.808g, 1mmol) was dissolved in 20ml dichloromethane, N-diisopropylethylamine (0.323g, 2.5mmol) was added, the temperature was reduced to 0 ℃, reaction solution 1 was dropped, and the reaction was continued for 2 hours. Washing the precipitate once with 30ml of saturated sodium dihydrogen phosphate aqueous solution, drying the precipitate with anhydrous sodium sulfate, concentrating the precipitate under reduced pressure, and performing column chromatography by using ethyl acetate/petroleum ether (v/v) of 0: 1-1: 1 as an eluent. Compound 41 was obtained as a white solid (0.1g, 8% yield).

¹H NMR(400MHz,CDCl₃)δ8.12–8.03(m,2H),7.60(d,1H),7.49(t,2H),7.45–7.35(m,4H),7.32(d,3H),7.22–7.15(m,2H),5.98(s,1H),5.63(d,1H),5.36(d,2H),5.33–5.20(m,3H),5.15–4.92(m,4H),4.30-4.16(m,3H),4.04–3.84(m,3H),3.84–3.57(m,3H),2.56-2.52(m,1H),2.36(s,3H),2.16(d,2H),1.98–1.85(m,2H),1.73(s,3H),1.41–1.35(m,15H),1.30–1.22(m,12H),1.13(s,3H),1.07(s,3H).

³¹P NMR(162MHz,CDCl₃)δ9.71.

Example 42

(2AR,4S,4AS,6R,9S,11S,12S,12AR,12bS) -12b- (acetoxy) -12- (benzoyloxy) -2a,3,4,4a,5,6,9,10,11,12,12a,12 b-dodecahydro-4, 6, 11-trihydroxy-4 a,8,13, 13-tetramethyl-5-oxo-7, 11-methylene-1H-cyclododecene [3,4] benzo [1,2-b ] oxetan-9-yl ((alpha R, beta S) -beta- [ [ (1, 1-dimethylethoxy) carbonyl ] amino ] -alpha-isopropyl (2S) -2- [ [ [ (1S) -2-isopropoxy-1-methyl-2-oxo-ethyl ] amino ] Phosphonylamino ] propionate-phenylpropionate (Compound 42)

(2aR,4S,4aS,6R,9S,11S,12S,12aR,12bS)-12b-(acetyloxy)-12-(benzoyloxy)-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-4,6,11-trihydroxy-4a,8,13,13-tetramethyl-5-oxo-7,11-methano-1H-cyclodeca[3,4]benz[1,2-b]oxet-9-yl(αR,βS)-β-[[(1,1-dimethylethoxy)carbonyl]amino]-α-isopropyl(2S)-2-[[[(1S)-2-isopropoxy-1-methyl-2-oxo-ethyl]amino]phosphonoylamino]propanoate-Benzenepropionate

Dissolving phosphorus oxychloride (0.154g, 1mmol) in 30ml dichloromethane, adding a mixed solution of triethylamine (0.203g, 2.01mmol) and docetaxel (4A) (0.808g, 1mmol) in dichloromethane dropwise under the protection of nitrogen at-30 ℃, continuing stirring for 30min after dropwise addition, adding L-alanine isopropyl ester hydrochloride (0.354g, 2.11mmol), adding triethylamine (0.407g, 4.02mmol), naturally raising the temperature to room temperature after addition, and stirring for 2 h. Washed once with 30ml of a saturated aqueous solution of sodium dihydrogenphosphate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by separation with a liquid phase preparative column to give compound 42 as a white solid (0.06g, yield 5%).

Liquid phase preparation conditions: the instrument comprises the following steps: gilson GX-281; column:

AD-H, 20 × 250mmi.d.,5 μm.; mobile phase: a for n-hexane and B for isoproyl alcohol; isocratic elution: a80 percent; flow rate: 12mL/min, back pressure: 1000 PSI; column temperature: 30 ℃; wavelength: 210 nm; and (3) period: 35 min; sample preparation: compound DXTS was dissolved in 12mL isopropanol; and (3) injection: 0.9 mL/needle.

¹H NMR(400MHz,CDCl₃)δ8.10(d,2H),7.58(d,1H),7.49(t,2H),7.40(t,2H),7.34(d,2H),7.27(d,1H),6.23(s,1H),5.66(d,1H),5.57(d,1H),5.40(s,1H),5.27(s,1H),5.20(s,1H),5.09-4.98(m,2H),4.94(d,1H),4.31-4.27(m,2H),4.19(d,1H),4.00(dd,1H),3.92(d,1H),3.73(t,1H),3.56(d,1H),2.96(s,1H),2.63-2.55(m,1H),2.43(s,3H),2.27(s,1H),1.98(d,4H),1.93–1.83(m,1H),1.73(s,4H),1.36(d,3H),1.32(s,9H),1.28-1.22(m,15H),1.12(s,3H),1.04(t,3H).

³¹P NMR(162MHz,CDCl₃)δ14.68.

Example 43

(2aR,4S,4aS,9S,11S,12S,12aR) -12 b-acetoxy-9- (((2R,3S) -3- ((tert-butyloxycarbonyl) amino) -2- ((((4- (((((((((S) -1-isopropyl-1-oxopropan-2-yl) amino) (methoxymethyl) phosphoryl) oxy) benzoyl) oxy) carbonyl) oxy) -3-phenylpropionyl) oxy) -4,6, 11-trihydroxy-4 a,8,13, 13-tetramethyl-5-oxo-2 a,3,4,4a,5,6,9,10,11,12,12a,12 b-dodecahydro-1H-7, 11-methanocyclododecyl [3,4] benzo [1,2-b ] oxetane-12-benzoate (Compound 43)

(2aR,4S,4aS,9S,11S,12S,12aR)-12b-acetoxy-9-(((2R,3S)-3-((tert-butoxycarbonyl)amino)-2-((((4-(((((S)-1-isopropoxy-1-oxopropan-2-yl)amino)(methoxymethyl)phosphoryl)oxy)benzyl)oxy)carbonyl)oxy)-3-phenylpropanoyl)oxy)-4,6,11-trihydroxy-4a,8,13,13-tetramethyl-5-oxo-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-1H-7,11-methanocyclodeca[3,4]benzo[1,2-b]oxet-12-yl benzoate

Intermediate 2(2.03g, 5.88mmol) was dissolved in 20mL dry tetrahydrofuran, cooled at-30 deg.C, triphosgene (0.88g,3mmol) and N, N-diisopropylethylamine (0.8g,6.19mmol) were added and stirred for 1h to give solution 1; docetaxel (5g, 6.19mmol) was dissolved in 30mL dry THF, cooled at-30 deg.C, and 1, 4-dimethylaminopyridine (0.72g,5.90mmol) and N, N-diisopropylethylamine (0.8g,6.19mmol) were added dropwise, slowly warmed to room temperature for 5h, and the reaction was stopped. Concentration and silica gel column chromatography (petroleum ether/ethyl acetate (v/v) ═ 2/3) gave compound 43 as a white solid, and compound 6 was further separated by liquid phase preparative column to give compound 43-a (1.7g) (peak 1, white solid, c-HPLC: 100%) and compound 43-b (3.2g) (peak 2, white solid, c-HPLC: 98.20%), with compounds 6-a and 6-b being diastereomers and the peak 1 showing a peak off time longer than peak 2.

The liquid phase preparation conditions were as follows: the instrument is Gilson GX-281; xbridge C18,150X 30mm I.D.,5 μm.; mobile phase A, acetonitrile, phase B: water (0.1% ammonium acetate); gradient, phase A is 45-55-15 min; the flow rate is 30 mL/min; the back pressure is 1200 PSI; the column temperature is 30 ℃; the wavelength is 210 nm; the period is 18 min.

Compound 43-a:

¹HNMR(400MHz,CDCl₃):δ8.80(d,2H),8.71(d,1H),8.53(t,1H),8.45(t,2H),8.24-8.15(m,6H),8.01-7.97(m,3H),6.60(t,1H),6.21(d,1H),5.99(t,2H),5.91-5.90(m,2H),5.85(d,1H),5.80(d,1H),5.72-5.64(m,3H),5.23(s,1H),4.90-4.80(m,4H),4.72-4.65(m,1H),4.60(d,2H),4.45(d,1H),4.20(s,3H),3.10-3.05(m,4H),2.68-2.62(m,1H),2.52-2.43(m,1H),2.38-2.33(m,4H),2.15(s,9H),2.07-2.03(m,1H),1.99-1.96(m,12H),1.80(s,6H).

³¹P NMR(162MHz,CDCl₃)δ25.83.

compound 43-b:

1HNMR(400MHz,CDCl3):δ8.80(d,2H),8.71(d,1H),8.53(t,1H),8.45(t,2H),8.24-8.15(m,6H),8.03(d,2H),7.98(t,1H),6.60(t,1H),6.21(d,1H),5.99(t,2H),5.92-5.90(m,2H),5.85(d,1H),5.80(d,1H),5.72-5.64(m,3H),5.23(s,1H),4.90-4.80(m,4H),4.71-4.64(m,1H),4.57(d,2H),4.45(d,1H),4.17(s,3H),3.10-3.03(m,4H),2.68-2.62(m,1H),2.53-2.43(m,1H),2.38-2.33(m,4H),2.15(s,9H),2.07-2.03(m,1H),2.00-1.96(m,12H),1.80(s,6H).

³¹P NMR(162MHz,CDCl₃)δ24.95.

biological test example

Mouse pharmacokinetic testing

Test animals: male ICR mice, about 25g, 6-8 weeks old. Purchased from Woods laboratories Inc. Animals were kept in an SPF environment at 20-22 ℃, relative humidity: 51-55%, 12h/12h light and dark illumination, drinking water freely, and starting the test after 3 days of adaptability observation.

Preparing the medicine: accurately weighing a certain amount of tested compound, dissolving the compound in DMSO, adding solutol HS-15 for solubilization, adding physiological saline, and mixing by vortex. The final concentration of DMSO was 5%, and the final concentration of solutol HS-15 was 5%. All compounds tested were prepared fresh just before use.

Administration and detection: on the day of the experiment, ICR mice were randomly grouped by body weight, 9 mice per group. The food is fasted for 12-14 h before administration for 1 day, and is fed for 4h after administration. Mice were given different test compounds orally, respectively, in a volume of 10 mL/kg. Before and after administration, isoflurane is anesthetized and blood is taken by 0.10ml through eye sockets, the blood taking time points are selected from 0min, 5min, 10min, 15min, 30min, 1h, 1.5h, 2h, 4h, 6h, 8h, 24h, 30h and 48h, heparin is anticoagulated, the mixture is centrifuged at 5000rpm for 10min at 4 ℃, and blood plasma is collected. All plasma samples were stored at-80 ℃ prior to analysis. The plasma sample neutralization proto-drug was tested by HPLC-MS/MS and the results are shown in Table 1 below:

TABLE 1

Pharmacokinetic testing of rats

Test animals: male SD rats, about 180-220 g, 6-8 weeks old, were purchased from Duoduoshuo laboratory animals Co., Ltd. Animals were kept in an SPF environment at 20-22 ℃, relative humidity: 51-55%, 12h/12h light and dark illumination, drinking water freely, and starting the test after 3 days of adaptability observation.

Administration and detection: on the day of the experiment, the SD rats were randomly divided into 4 groups of 3 rats by body weight. The food is fasted for 12-14 h before administration for 1 day, and is fed for 4h after administration. Rats were given different test compounds intravenously or orally, respectively, in a volume of 10 mL/kg. Before and after administration, isoflurane is anesthetized and blood is taken by 0.20ml through eye sockets, the blood taking time points are selected from 0min, 5min, 10min, 15min, 30min, 1h, 1.5h, 2h, 4h, 6h, 8h, 24h, 30h and 48h, heparin is used for anticoagulation, 6000rpm is adopted, centrifugation is carried out for 10min at 4 ℃, and blood plasma is collected. All plasma samples were stored at-80 ℃ prior to analysis. The results of the detection of the proto-drug in plasma samples by HPLC-MS/MS are shown in Table 2 below:

TABLE 2

N/A: no blood drug and no absorption can be detected by oral administration of docetaxel rats.

Beagle pharmacokinetic testing

Test animals: beagle dogs, 6-8 kg males, were purchased from Duoduoshu laboratory animals, Inc. Animals were kept in an SPF environment at 20-22 ℃, relative humidity: 51-55%, 12h/12h light and dark illumination, drinking water freely, and starting the test after 3 days of adaptability observation.

Administration and detection: on the day of the experiment, beagle dogs were randomly grouped by weight, 3 dogs each. The food is fasted for 12-14 h before administration for 1 day, and is fed for 4h after administration. Beagle dogs were given different test compounds orally in a volume of 5 mL/kg. Taking 1.5ml of blood through veins of limbs before and after administration, selecting certain time of 0min, 5min, 10min, 15min, 30min, 1h, 1.5h, 2h, 4h, 6h, 8h, 10h,12h,24h, 30h and 48h for blood taking time points, anticoagulating heparin, centrifuging at 3500rpm for 10min at 4 ℃, and collecting plasma. All plasma samples were stored at-80 ℃ prior to analysis. The prototype drug in the plasma sample is detected by HPLC-MS/MS, and the result shows that the test compound has good oral performance.

TABLE 3 Biggee pharmacokinetic parameters

Claims

1. A compound of formula (I) and stereoisomers or pharmaceutically acceptable salts thereof, wherein the compound is selected from one of the following structures:

2. a compound of formula (I) and stereoisomers or pharmaceutically acceptable salts thereof according to claim 1, wherein the compound is selected from one of the following structures:

3. a compound of formula (I) and stereoisomers or pharmaceutically acceptable salts thereof according to claim 1 or 2, wherein the compound is selected from one of the following structures:

4. a pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1to 3, or a stereoisomer or a pharmaceutically acceptable salt thereof, together with pharmaceutically acceptable carriers and excipients.

5. A compound according to any one of claims 1to 3, a stereoisomer or a pharmaceutically acceptable salt thereof, and a composition according to claim 4 for use in the preparation of a medicament for the prevention or treatment of diabetes, anti-tumor, hyperkinetic movement disorders, motor neuron diseases, cerebral protection, diseases related to the central nervous system, immunosuppression, rejection by organ tissue or cell xenosuppression, immunomodulation and/or inflammatory diseases, convulsions, epilepsy or stroke.

6. The use according to claim 5, wherein the diabetes is selected from type II diabetes, the hyperkinetic movement disorder is selected from chorea, Huntington's disease, Tourette's disease, tardive dyskinesia or unilateral tic movement, the neoplasm is selected from prostate cancer, pancreatic cancer, lung cancer, ovarian cancer, cervical cancer, liver cancer, breast cancer, esophageal cancer, stomach cancer, colorectal cancer, cholangiocarcinoma, nasopharyngeal cancer, testicular cancer, lymphatic disease, head and neck cancer, squamous cell carcinoma, colorectal cancer, leukemia, brain cancer, neck tumor and liver cancer, hematological cancer or bladder cancer, the motor neuron disease is selected from amyotrophic lateral sclerosis, and the central nervous system-related disease is selected from multiple sclerosis.