CN111393399A

CN111393399A - Prodrug of mycophenolic acid and preparation method thereof

Info

Publication number: CN111393399A
Application number: CN202010419146.5A
Authority: CN
Inventors: 王延东; 刘国强; 刘伟
Original assignee: Shanghai Innofucheng Biotechnology Co ltd
Current assignee: Guangzhou Ocusun Ophthalmic Biotechnology Co Ltd
Priority date: 2019-08-20
Filing date: 2020-05-18
Publication date: 2020-07-10
Anticipated expiration: 2040-05-18
Also published as: CN111393399B; WO2021031999A1; CN110526885A

Abstract

The invention belongs to the field of medicines, and particularly relates to a prodrug of mycophenolic acid and a preparation method thereof. The structural general formula is shown as formula I: compared with mycophenolic acid, the compound shown in the formula I has better aqueous permeability, stability of aqueous solution and better water solubility, and can be hydrolyzed into mycophenolate mofetil in aqueous humor, so that the medicine concentration of mycophenolic acid in aqueous humor is obviously improved compared with that of mycophenolate mofetil, and the medicine concentration is observed to be 2 times or more higher than that of mycophenolate mofetil in experiments, and the compound is more suitable for preparing eye liquidAnd (4) preparing the preparation.

Description

Prodrug of mycophenolic acid and preparation method thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to a prodrug of mycophenolic acid and a preparation method thereof.

Background

Mycophenolate Mofetil (MMF) is a potent, novel immunosuppressive drug which is the 2-morpholino ethylation product of mycophenolic acid (mycophenolic acid, MPA). Inosine monophosphate dehydrogenase is an enzyme required in the synthesis of guanine nucleotide by lymphocytes and is thus essential for DNA synthesis and cell proliferation. MMF is a noncompetitive, reversible inhibitor of this enzyme. Inhibition of hypoxanthine monophosphate dehydrogenase by MPA results in cell retention in the G1 phase of the cell cycle. In T cells, MPA inhibits the activity of Cyclin Dependent Kinases (CDKs), blocking the clearance of cDK inhibitor p27 (Kipl). In addition to its effect on lymphocyte proliferation, MMF can also inhibit glycosylation of glycoproteins in lymphocytes and monocytes that are associated with adhesion to endothelial cells. Meclofenol sodium enteric-coated tablets, suitable for use in combination with cyclosporin and a corticosteroid, for the prevention of acute rejection in adult patients receiving allogeneic kidney transplants.

The mycophenolate mofetil is clinically used in two types of tablets and injections, and because of poor solubility, polysorbate 80 is required to be added and used as a cosolvent in the injections. The marketed dosage forms of mecofenphen sodium are capsules and tablets. The aqueous solution of mycophenolate mofetil and mycophenolate sodium is unstable, and is hydrolyzed into mycophenolic acid in the aqueous solution, and phenolic hydroxyl is easily oxidized into unknown impurities. The above factors make mycophenolate mofetil and mycophenolate sodium unable to be directly made into liquid preparation (injection, oral solution, eye drops, etc.).

Disclosure of Invention

In order to solve the technical problems, the invention provides a compound.

The structural general formula of the compound provided by the invention is shown as formula I:

in the above formula I, R₁、R₂Can be independently selected from: -NH₂H, amino/hydroxy/halogen/aryl/carboxy substituted C₁-C₁₀Alkyl (specifically amino/hydroxy/halogen/aryl/carboxy substituted C)₁-C₅Alkyl radicals, e.g. C₆H₅CH₂-、-CH₂CH₂COOH), amino/hydroxy/halogen substituted C₃-C₆Cycloalkyl radical, C₁-C₁₀Alkyl (which may be C in particular)₁-C₅Alkyl radicals, e.g. -CH₂CH₂CH₂CH₃、-CH₃)、C₃-C₆Cycloalkyl, -N (CH)₂CH₃)₂、-N(CH₃)₂Any one of the above;

R₃is H or-CH₃，

R₃When is H, R₁、R₂Not both can be-H; r₃When is H, R₁、R₂Not one being H and the other being-NH₂；

R₄、R₅Can be independently selected from: -H, C₁-C₅Alkyl (which may be C in particular)₁-C₃Alkyl radicals, e.g. -CH₃、-CH₂CH₃) Any one of the above;

x represents O or N;

n is an integer of 1 to 8, specifically 1 to 5 or 1 to 3, more specifically 1 or 2.

In some embodiments, the compounds of the present invention may be exemplified by, but are not limited to, the structures shown below:

the compound shown in the formula I is prepared by a method comprising the following steps:

carrying out condensation reaction on a compound shown in a formula II and a compound shown in a formula III to obtain a compound shown in a formula I;

in the formula II R₄、R₅X is the same as R in formula I₄、R₅And X, n is an integer of 1-8, and can be 1 specifically;

in the formula III, R₁、R₂、R₃In the same formula I as R₁、R₂、R₃The definition of (1);

in the above method, the condensation reaction is carried out in the presence of an acid-binding agent;

the acid-binding agent can be DIPEA, triethylamine, pyridine and the like, and specifically can be DIPEA;

the mol ratio of the compound shown in the formula II to the compound shown in the formula III and the acid-binding agent can be: 1:1-2:2-4, specifically 1:1.5: 3;

the condensation reaction is carried out in an organic solvent, and the organic solvent can be DCM;

the reaction temperature can be room temperature, and the reaction time can be 8-14 h.

It is a further object of the present invention to provide the use of a compound of formula I as described above, or a pharmaceutically acceptable salt, ester, solvate or isomer (including stereoisomers, enantiomers, tautomers or mixtures thereof).

The salt can be hydrochloride or tartaric acid.

The application provided by the invention comprises the following aspects:

the use of a compound of formula I or a pharmaceutically acceptable salt, ester, solvate or isomer (including stereoisomers, enantiomers, tautomers or mixtures thereof) thereof as a prodrug of mycophenolic acid;

the use of a compound of formula I or a pharmaceutically acceptable salt, ester, solvate or isomer (including stereoisomers, enantiomers, tautomers or mixtures thereof) thereof in the preparation of an immunosuppressant;

specifically, the application is the application of the compound shown in the formula I or pharmaceutically acceptable salts, esters, solvates or isomers (including stereoisomers, enantiomers, tautomers or mixtures thereof) thereof in preparing a medicament for preventing or treating transplant rejection and immune-mediated and/or inflammatory diseases.

The invention also provides a pharmaceutical formulation comprising a compound of formula I, or a pharmaceutically acceptable salt, ester, solvate or isomer thereof, and one or more of: excipient, diluent, pH stabilizer.

The pharmaceutical preparation can be prepared into various dosage forms including liquid preparations.

The liquid preparation can be at least one of injection, oral solution and eye liquid preparation.

The liquid formulation does not add a co-solvent including polysorbate 80.

The invention also provides an eye liquid preparation, which comprises the compound shown in the formula I or pharmaceutically acceptable salt, ester, solvate or isomer thereof, a solvent and one or more physiologically acceptable auxiliary materials.

In the ophthalmic liquid preparation, the solvent is water.

The ophthalmic liquid preparation does not add a cosolvent including polysorbate 80.

Compared with mycophenolic acid, mycophenolate sodium and mycophenolate mofetil, the compound shown by the general formula of the invention has the following advantages: 1. the water solubility is better; 2. compared with sodium mycophenolate and mycophenolate mofetil, the mycophenolate mofetil is stable in a system containing water components and hardly has degradation; 3. has good permeability, can permeate cornea and enter aqueous humor, has higher aqueous humor drug concentration which is 2 to 5 times of mycophenolic acid; 4. the aqueous humor can be hydrolyzed into mycophenolic acid to exert the drug effect.

The compound shown in the formula I has better aqueous humor permeability than mycophenolic acid, so that the medicinal concentration of the mycophenolic acid in the aqueous humor is obviously improved compared with that of mycophenolate sodium, and experiments show that the medicinal concentration of the compound shown in the examples 1-11 in the aqueous humor of the New Zealand rabbit after administration is 2 times or more higher than that of the mycophenolate sodium, so that the compound is more suitable for preparing an eye liquid preparation.

Drawings

FIG. 1 is a scheme for preparing a target compound in example 1 of the present invention.

FIG. 2 is a scheme for preparing a target compound in example 2 of the present invention.

FIG. 3 is a scheme showing the preparation of a target compound in example 3 of the present invention.

FIG. 4 is a scheme showing the preparation of the objective compound in example 4 of the present invention.

FIG. 5 is a scheme showing the preparation of the objective compound in example 5 of the present invention.

FIG. 6 is a scheme showing the preparation of a target compound in example 6 of the present invention.

FIG. 7 is a scheme showing the preparation of a target compound in example 7 of the present invention.

FIG. 8 is a scheme showing the preparation of a target compound in example 8 of the present invention.

FIG. 9 is a scheme showing the preparation of a target compound in example 9 of the present invention.

FIG. 10 is a scheme showing the preparation of a target compound in example 11 of the present invention.

FIG. 11 is a scheme showing the preparation of a target compound in example 12 of the present invention.

Detailed Description

The present invention will be described below with reference to specific examples, but the present invention is not limited thereto.

The experimental methods used in the following examples are all conventional methods unless otherwise specified; reagents, materials and the like used in the following examples are commercially available unless otherwise specified.

If the sodium mycophenolate or the derivative of mycophenolate needs to be prepared into a stable eye liquid preparation, three conditions need to be met: 1. can be decomposed into mycophenolic acid in aqueous humor; 2. can be kept stable in aqueous solution; 3. has good water solubility, and can be prepared into aqueous solution preparation.

A series of compounds are designed from mycophenolate sodium, and an ocular pharmacokinetic experiment and a stability experiment show that the compounds with the following general formula can meet the requirements, can be prepared into an ocular liquid preparation to be used as a prodrug of mycophenolic acid, and can exert the drug effect of mycophenolic acid locally. R₁、R₂、R₃The selection of (A) plays a critical role in the stability and ability to hydrolyze in aqueous humor to mycophenolic acid, as observed in experiments when R is₁Is H, R₂Is H, R₃When the compound is H, the compound shown in the formula I cannot be stably kept in an aqueous solution and is easy to hydrolyze, and the phenolic hydroxyl group after hydrolysis generates oxidation reaction, R₁Is H, R₂Is NH₂,R₃A similar situation occurs for H. The compound shown in the formula I has better aqueous humor permeability than the mycophenolic acid, so that the medicine concentration of the mycophenolic acid in the aqueous humor is obviously improved compared with the mycophenolate sodium, the medicine concentration is observed to be 3 times or more than that of the mycophenolate sodium in the experiment, the compound is more suitable for preparing the eye liquid preparation,

in the above formula I, R₁、R₂Can be independently selected from: -NH₂H, amino/hydroxy/halogen/aryl/carboxy-substituted C₁-C₁₀Alkyl (specifically amino/hydroxy/halogen/aryl/carboxy substituted C)₁-C₅Alkyl radicals, e.g. C₆H₅CH₂-、-CH₂CH₂COOH), amino/hydroxy/halogen substituted C₃-C₆Cycloalkyl radical, C₁-C₁₀Alkyl (which may be C in particular)₁-C₅Alkyl radicals, e.g. -CH₂CH₂CH₂CH₃、-CH₃)、C₃-C₆Cycloalkyl, -N (CH)₂CH₃)₂、-N(CH₃)₂Any one of the above;

R₃is H or-CH₃，

x represents O or N;

n is an integer of 1 to 8, and specifically may be 1.

Examples 1,

Preparation of (SZY1805-1H) Compound

The above compounds were prepared according to the synthetic scheme shown in figure 1:

adding SZY-1805-2H (10.0g, 31.22mmol, 1.0eq, MW320.34) into a 250ml round bottom flask, adding 100ml of DMF for dissolution, adding imidazole (13.8g, 202.9mmol, 6.5eq, MW68.08) for dissolution, then adding TBSCl (18.82g, 124.87mmol, 4.0eq, MW150.7) in portions, stirring at room temperature overnight, detecting the completion of the reaction by T L C, adding 150ml of EA and 200ml of water layer into the solution, extracting the water layer three times by 50ml of EA, combining the organic phase, washing the water layer by 1% hydrochloric acid solution, washing by pure water 3-5 times (removing DMF), drying the organic phase by anhydrous sodium sulfate, evaporating to obtain an oily substance, adding THF50 oily ml, water 50ml, acetic acid 50ml, stirring the reaction by normal temperature, after 2H, monitoring the water layer by T L C, adding 150ml of EA and 200ml of water into the reaction solution, adding 150ml of rotary extraction solution, drying the white solid (adding water layer), extracting by evaporation, adding white solid, drying by suction filtration, obtaining white solid, mixing the white solid, obtaining white solid, adding 464 g, adding water, drying, adding 464 g and 4688 g of water, adding water, after filtration to obtain white solid;

to a 250ml round bottom flask was added SZY-1805-4H (5.0g, 11.5mol, 1.0eq, MW434.6) and dissolved by the addition of 50ml dichloromethane. Add BocNHC sequentially₂H₄OH(2.22g，13.8mmol，1.2eq，

MW161.2), DMAP (150mg, 1.2mmol, 0.1eq, MW122), EDCI (2.6g, 13.8mmol, 1.2eq, MW191.7), reaction 7 h.T L C was stirred at room temperature to monitor completion of the reaction, DCM was removed by rotary evaporation under reduced pressure, EA150ml was added, water was separated into layers 150ml, the organic phase was washed 2 times with 100ml of water, 2 times with 100ml of saturated brine, dried over anhydrous sodium sulfate, filtered and rotary evaporated to give 6.05g of oil in 91% yield (SZY-1805-5H).

To a 250ml round bottom flask containing SZY-1805-5H (6.05g, 10.47mmol, 1.0eq, MW577.8) was added 50ml thf to dissolve, followed by TBAF (3.96g, 12.56mmol, 1.2eq, MW315.5) and the reaction stirred at room temperature, TBAF was added and the solution turned wine red and then orange yellow, after 3H T L C monitored, the reaction was completed, water and EA were added each 100ml, the layers were separated, the organic phase was washed with water, dried over anhydrous sodium sulfate, rotary evaporated, and purified by column chromatography to give 5.33g (containing solvent) as an oil in 110% yield with a small amount of solvent (SZY-1805-6H).

SZY-1805-6H (5.33g, 11.5mmol, 1.0eq, MW463.52) is dissolved in 50ml DCM, DIPEA (4.46g, 34.5mmol, 3eq, MW129.25) is added to the DCM, stirring is carried out for reaction, propionyl chloride (1.60g, 17.5mmol, 1.5eq, MW92.52) is added in portions at 0 ℃, about half an hour is finished, after the end of the valence-lowering reaction, the reaction solution is heated to room temperature and stirred overnight, T L C monitors the end of the reaction on the next day, the reaction solution is washed 2 times with 100ml of water, 100ml of saturated saline solution is washed 2 times, the organic phase is dried with anhydrous sodium sulfate, rotary evaporation is carried out, column chromatography is carried out, and 5.25g of oily liquid is obtained, and the yield is 88% (SZY-1805-7H).

And (2) dissolving SZY-1805-7H (5g) in 20ml of EA, adding 4M HCl 50ml of EA into the solution, stirring the solution at normal temperature for reaction, monitoring the reaction at intervals of half an hour by T L C, removing the solvent by rotary evaporation after the reaction is finished to obtain an oily substance, adding a sodium bicarbonate aqueous solution to neutralize hydrochloric acid, extracting the hydrochloric acid by ethyl acetate, drying the anhydrous sodium sulfate, and evaporating the solvent to obtain 3g of a product (SZY 1805-1H).

The structure identification data is as follows:

1H NMR:SZY1805-1H(400MHz CDCl3)8.36(s，2H)，5.16(s，2H)，5.08-5.06(t，1H)，4.32(s，2H)，3.78(s，3H)，3.33-3.22(m，4H)，2.73-2.68(m，2H)，2.49-2.46(m，2H)，2.30-2.26(m，2H)，2.22(s，3H)，1.76(s，3H)，1.32-1.24(m，3H)。MS：[M+H]+＝420.0。

examples 2,

Preparation of (SZY1805-1a) Compound

The above compounds were prepared according to the synthetic scheme shown in figure 2:

charging SZY-1805-2a (10.0g, 31.22mmol, 1.0eq, MW320.34) into a 250ml round bottom flask, charging 100ml DMF for dissolution, charging imidazole (13.8g, 202.9mmol, 6.5eq, MW68.08) for dissolution, then charging TBSCl (18.82g, 124.87mmol, 4.0eq, MW150.7) in portions, stirring overnight at room temperature, detecting completion of reaction by T L C, charging 150ml EA and 200ml water layer into the solution, extracting the water phase three times with 50ml EA, combining the organic phase, washing the water phase with 1% hydrochloric acid solution, washing with pure water 3-5 times (removing DMF), drying the organic phase with anhydrous sodium sulfate, evaporating to obtain rotary, charging THF50 oily ml, water 50ml, acetic acid 50ml, stirring for reaction, monitoring reaction water content after 2h, charging 150ml EA and 200ml EA into the reaction solution, charging 150ml EA and 200ml water layer into the reaction solution, mixing the white organic phase with yellow water layer, drying, adding yellow silica, extracting solvent, filtering to obtain white oily ml, mixing the white oily 3970 g, filtering to obtain white solid, filtering to obtain filtrate, filtering to obtain white solid, filtering to obtain filtrate, filtering.

To a 250ml round bottom flask was added SZY-1805-3a (5.0g, 11.5mol, 1.0eq, MW434.6) and dissolved by the addition of 50ml dichloromethane. SZY-1805-4a (1.5g, 13.8mmol, 1.2eq,

MW117.2), DMAP (150mg, 1.2mmol, 0.1eq, MW122), EDCI (2.6g, 13.8mmol, 1.2eq, MW191.7), reaction 7 h.T L C was stirred at room temperature to monitor completion of the reaction, DCM was removed by rotary evaporation under reduced pressure, EA150ml was added, water was separated into layers, the organic phase was washed 2 times with 100ml of water, 2 times with 100ml of saturated brine, dried over anhydrous sodium sulfate, and rotary evaporated by filtration to give 5.8g of oil in 99% yield (SZY-1805-5 a).

To a 250ml round bottom flask containing SZY-1805-5a (5.8g, 11.5mmol, 1.0eq, MW533.8) was added 50ml THF to dissolve, then TBAF (4.4g, 13.8mmol, 1.2eq, MW315.5) was added and the reaction stirred at room temperature, the solution turned wine red after TBAF addition and then turned orange yellow, after 3h T L C monitor, reaction was completed, 100ml each of water and EA was added thereto, the organic phase was washed with water, dried over anhydrous sodium sulfate, rotary evaporated, and column chromatography purification gave 4.0g of oil (containing solvent) with 108% yield (SZY-1805-6 a).

SZY-1805-6a (4.0g, 10.49mmol, 1.0eq, MW419.5) is dissolved in 50ml DCM, DIPEA (4.07g, 31.47mmol, 3eq, MW129.25) is added to the solution, the reaction is stirred, propionyl chloride (1.45g, 15.73mmol, 1.5eq, MW92.52) is added in portions at 0 ℃, about half an hour is finished, after the end of the reaction, the reaction is stirred overnight at room temperature, the reaction solution is monitored by T L C for the end of the reaction, the reaction solution is washed 2 times with 100ml of water, the saturated saline solution is washed 2 times with 100ml, the organic phase is dried with anhydrous sodium sulfate, rotary evaporated, and separated by column chromatography, and 3.0g of oily liquid is obtained with the yield of 65% (SZY1805-1 a).

The structure identification data is as follows:

1H NMR:SZY1805-1a(400MHz CDCl3)5.17-5.15(d，J＝9.6Hz，2H)，5.08-5.05(t，1H)，4.52-4.49(m，2H)，3.80(s，3H)，3.23-3.12(m，2H)，3.12-3.11(m，2H)，3.05-2.86(m，6H)，2.74-2.68(m，2H)，2.46-2.40(m，2H)，2.30-2.26(m，2H)，2.24(s，3H)，1.77(s，3H)，1.40-1.36(m，3H)。MS：[M+H]+＝448.5。

examples 3,

Preparation of (SZY1805-1f) compound the above compound was prepared according to the synthetic scheme shown in figure 3:

charging SZY-1805-2f (10.0g, 31.22mmol, 1.0eq, MW320.34) into a 250ml round bottom flask, charging 100ml DMF for dissolution, charging imidazole (13.8g, 202.9mmol, 6.5eq, MW68.08) for dissolution, then charging TBSCl (18.82g, 124.87mmol, 4.0eq, MW150.7) in portions, stirring overnight at room temperature, detecting completion of reaction by T L C, charging 150ml EA and 200ml water layer into the solution, extracting the water phase three times with 50ml EA, combining the organic phase, washing the water phase with 1% hydrochloric acid solution, washing with pure water 3-5 times (DMF removal), drying the organic phase with anhydrous sodium sulfate, evaporating to obtain an oily substance, charging THF50 oily substance, water 50ml, acetic acid 50ml, stirring for reaction, monitoring reaction water content after 2h, charging 150ml EA and 200ml EA into the reaction solution, charging 150ml EA into the reaction solution, charging 200ml water layer, evaporating to obtain a white oily substance, drying (PE) to obtain a white oily substance, adding water, filtering to obtain a white oily substance, drying after 2h, adding 150ml, stirring reaction, filtering to obtain a white oily substance, filtering to obtain a white solid, and filtering to obtain a filtrate.

To a 250ml round bottom flask was added SZY-1805-3f (5.0g, 11.5mol, 1.0eq, MW434.6) and dissolved by addition of 50ml dichloromethane SZY1805-4f (1.6g, 13.8mmol, 1.2eq, MW117.2), DMAP (150mg, 1.2mmol, 0.1eq, MW122), EDCI (2.6g, 13.8mmol, 1.2eq, MW191.7) were added in sequence, reaction 7 h.T L C was stirred at room temperature to monitor completion of the reaction, DCM was removed by reduced pressure rotary evaporation, EA150ml was added, water was layered 150ml, organic phase water was washed 2 times with 100ml brine saturated for 2 times, dried over anhydrous sodium sulfate, filtered rotary evaporated to give 6.1g of oil in 99% yield (SZY-1805-5 f).

To a 250ml round bottom flask containing SZY1805-5f (6.1g, 11.5mmol, 1.0eq, MW533.8) was added 50ml thf to dissolve, followed by TBAF (4.4g, 13.8mmol, 1.2eq, MW315.5) and the reaction stirred at room temperature, TBAF was added to give a reddish-wine color and then a yellow-orange color, T L C was monitored after 3h, and after completion of the reaction, water and EA were added each 100ml, the organic phase was washed with water, dried over anhydrous sodium sulfate, rotary evaporated, and purified by column chromatography to give 4.4g (containing solvent) of oil in 108% yield (SZY-1805-6 f).

SZY-1805-6f (4.4g, 10.49mmol, 1.0eq, MW419.5) was dissolved in 50ml DCM, DIPEA (4.07g, 31.47mmol, 3eq, MW129.25) was added to it, the reaction was stirred, propionyl chloride (1.45g, 15.73mmol, 1.5eq, MW92.52) was added in portions at 0 deg.C, half an hour or so was completed, after completion of the lower valence, the reaction was stirred overnight at room temperature, the reaction was monitored for completion by T L C the next day, the reaction was washed 2 times with 100ml of water, 100ml of saturated saline was washed 2 times, the organic phase was dried over anhydrous sodium sulfate, rotary evaporated, column chromatographed, 3.22g of oily liquid was obtained, yield was 65% (SZY1805-1 f).

The structure identification data is as follows:

1H NMR:SZY1805-1f(400MHz CDCl3)5.17-5.15(d，J＝9.6Hz，2H)，5.08-5.05(t，1H)，4.52-4.49(m，2H)，3.80(s，3H)，3.23-3.12(m，4H)，3.12-3.11(m，4H)，2.74-2.68(m，2H)，2.46-2.40(m，2H)，2.30-2.26(m，2H)，2.24(s，3H)，1.77(s，3H)，1.46-1.44(m，6H)，1.40-1.36(m，3H)。MS：[M+H]+＝476.0。

examples 4,

Preparation of (SZY1805-1b) Compound

The above compounds were prepared according to the synthetic scheme shown in figure 4:

charging SZY-1805-2b (10.0g, 31.22mmol, 1.0eq, MW320.34) into a 250ml round bottom flask, charging 100ml DMF for dissolution, charging imidazole (13.8g, 202.9mmol, 6.5eq, MW68.08) for dissolution, then charging TBSCl (18.82g, 124.87mmol, 4.0eq, MW150.7) in portions, stirring overnight at room temperature, detecting completion of reaction by T L C, charging 150ml EA and 200ml water layer into the solution, extracting the water phase three times with 50ml EA, combining the organic phase, washing the water phase with 1% hydrochloric acid solution, washing with pure water 3-5 times (DMF removal), drying the organic phase with anhydrous sodium sulfate, evaporating to obtain an oil, charging THF50 oily ml, water 50ml, acetic acid 50ml, stirring for reaction, monitoring reaction water content after 2h, charging 150ml EA and 200ml EA into the reaction solution, charging 150ml EA into the reaction solution, charging 200ml water layer, evaporating to obtain white oil phase, drying (PE 5g, adding yellow, yellow wine, filtering to obtain white oil, drying, 2h, stirring to obtain white oil, filtering to obtain white solid, filtering.

To a 250ml round bottom flask was added SZY-1805-3b (5.0g, 11.5mol, 1.0eq, MW434.6) and dissolved by the addition of 50ml dichloromethane. SZY-1805-4b (1.6g, 13.8mmol, 1.2eq,

MW117.2), DMAP (150mg, 1.2mmol, 0.1eq, MW122), EDCI (2.6g, 13.8mmol, 1.2eq, MW191.7), reaction 7 h.T L C with stirring at room temperature, monitoring the completion of the reaction, removing DCM by rotary evaporation under reduced pressure, adding EA150ml, layering with 150ml of water, washing 2 times with 100ml of organic phase water, washing 2 times with 100ml of saturated brine, drying over anhydrous sodium sulfate, filtering and rotary evaporating to give 6.1g of oil in 99% yield (SZY-1805-5 b).

To a 250ml round bottom flask containing SZY1805-5b (6.1g, 11.5mmol, 1.0eq, MW533.8) was added 50ml THF to dissolve, then TBAF (4.4g, 13.8mmol, 1.2eq, MW315.5) was added and the reaction stirred at room temperature, the solution turned wine red after TBAF addition and then turned orange yellow, T L C was monitored after 3h, 100ml each of EA was added, the layers were separated, the organic phase was washed with water, dried over anhydrous sodium sulfate, rotary evaporated, and purified by column chromatography to give 4.4g (containing solvent) of oil with 108% yield (SZY-1805-6 b).

SZY-1805-6b (4.4g, 10.49mmol, 1.0eq, MW419.5) is dissolved in 50ml DCM, DIPEA (4.07g, 31.47mmol, 3eq, MW129.25) is added to the DCM, the reaction is stirred, dimethylaminochloride (1.6g, 15.73mmol, 1.5eq, MW92.52) is added in portions at 0 ℃, half an hour is finished, after the end of the reaction, the reaction is stirred overnight at room temperature, the reaction is monitored by T L C on the next day, the reaction solution is washed 2 times with 100ml of water, the saturated saline water is washed 2 times with 100ml, the organic phase is dried over anhydrous sodium sulfate, rotary evaporated, and separated by column chromatography, to obtain 3.5g of oily liquid with a yield of 70% (SZY1805-1 b).

The structure identification data is as follows:

1H NMR:SZY1805-1b(400MHz CDCl3)5.17-5.15(d，J＝9.6Hz，2H)，5.08-5.05(t，1H)，4.32-4.29(m，2H)，3.80(s，3H)，3.30-3.26(m，2H)，3.23-3.12(m，4H)，3.12-3.11(m，2H)，2.74-2.68(m，2H)，2.46-2.40(m，2H)，2.26(s，3H)，2.24(s，3H)，3.30-3.26(m，2H)，2.24(s，3H)，1.77(s，3H)，1.46-1.44(m，6H)。MS：[M+H]+＝405.6。

examples 5,

(SZY1805-1c) Preparation of the Compounds

The above compounds were prepared according to the synthetic scheme shown in figure 5:

SZY-1805-6(10.0) was added to a 250ml round bottom flask, and dissolved in 50ml of dichloromethane, SZY-1805-7C (5g), DMAP (0.2g) and EDCI (5g) were added in this order, the reaction was monitored at room temperature with stirring at 7 h.T L C, DCM was removed by rotary evaporation under reduced pressure, EA150ml was added, water was added and the reaction was separated into layers, the organic phase was washed 2 times with 100ml of water, 2 times with 100ml of saturated brine, dried over anhydrous sodium sulfate and rotary evaporated by filtration to give 8.0g of oil (SZY-1805-8C).

And (2) dissolving SZY-1805-8C (8.0g) in 40ml of EA, adding 4M HCl 40ml of EA, stirring at normal temperature for reaction, monitoring every half hour by T L C, removing the solvent by rotary evaporation after the reaction is finished to obtain an oily substance, adding a sodium bicarbonate aqueous solution to neutralize hydrochloric acid, extracting with ethyl acetate, drying with anhydrous sodium sulfate, and evaporating the solvent to obtain 6.2g of a product (SZY 1805-1C).

The structure identification data is as follows:

1H NMR:SZY1805-1c(400MHz CDCl3)5.17-5.15(d，J＝9.6Hz，2H)，5.08-5.05(t，1H)，4.52-4.49(m，2H)，3.80(s，3H)，3.61-3.57(m，1H)3.23-3.12(m，4H)，3.12-3.11(m，4H)，2.74-2.68(m，2H)，2.46-2.40(m，2H)，2.30-2.26(m，2H)，2.24(s，3H)，1.77(s，3H)，1.46-1.44(m，4H)，1.30-1.26(m，3H)。MS：[M+H]+＝491.6。

examples 6,

Preparation of (SZY1805-1d) Compound

The above compounds were prepared according to the synthetic scheme shown in figure 6:

SZY-1805-6(10g) is dissolved in 50ml DCM, DIPEA (4.07g, 31.47mmol, 3eq, MW129.25) is added into the DCM, stirring reaction is carried out, tert-valeryl chloride (2.0g) is added in batches at 0 ℃, the addition is completed by about half an hour, after the completion of the low price, the reaction is stirred overnight after the room temperature is raised to room temperature, the reaction is monitored to be completed on the next day by T L C, the reaction solution is washed for 2 times by 100ml of water, washed for 2 times by 100ml of saturated saline solution, an organic phase is dried by anhydrous sodium sulfate, rotary evaporation and column chromatography separation are carried out, and 8.0g of oily liquid is obtained with the yield of 80% (SZY1805-1 d).

The structure identification data is as follows:

1H NMR:SZY1805-1d(400MHz CDCl3)5.18(s，2H)，5.08-5.05(t，1H)，4.52-4.49(m，2H)，3.80(s，3H)，3.23-3.12(m，4H)，3.12-3.11(m，4H)，2.74-2.68(m，2H)，2.36-2.29(m，2H)，2.24(s，3H)，1.77(s，3H)，1.46-1.44(m，3H)，1.40-1.36(m，3H)1.2(s，9H)。MS：[M+H]+＝504.6。

example 7,

Preparation of (SZY1805-1e) Compound

The above compounds were prepared according to the synthetic scheme shown in figure 7:

SZY-1805-6(10.0) was added to a 250ml round bottom flask, and dissolved in 50ml of dichloromethane, SZY-1805-7e (6g), DMAP (0.2g), EDCI (5g) were added in this order, the reaction was stirred at room temperature to 12 h.T L C to monitor completion of the reaction, DCM was removed by rotary evaporation under reduced pressure, EA150ml was added, water 150ml was added and the reaction was separated into layers, the organic phase was washed 2 times with 100ml of water, 2 times with 100ml of saturated brine, dried over anhydrous sodium sulfate, and rotary evaporation was filtered to obtain 9.0g of an oil (SZY-1805-8 e).

And (3) dissolving SZY-1805-8e (9.0g) in 50ml of EA, adding 4M HCl 50ml of EA, stirring at normal temperature for reaction, monitoring every half hour by T L C, removing the solvent by rotary evaporation after the reaction is finished to obtain an oily substance, adding a sodium bicarbonate aqueous solution to neutralize hydrochloric acid, extracting with ethyl acetate, drying with anhydrous sodium sulfate, and evaporating the solvent to obtain 6.0g of a product (SZY1805-1 e).

The structure identification data is as follows:

1H NMR:SZY1805-1e(400MHz CDCl3)7.23-7.21(m，2H)，7.15-7.12(m，2H)，7.08-7.05(m，1H)，5.17-5.15(d，J＝9.6Hz，2H)，5.08-5.05(t，1H)，4.52-4.49(m，2H)，3.90-3.85(m，1H)，3.80(s，3H)，3.23-3.12(m，4H)，3.16-2.91(m，2H)，2.88-2.86(m，4H)，2.74-2.68(m，2H)，2.46-2.40(m，2H)，2.24(s，3H)，1.77(s，3H)，1.46-1.44(m，3H)，1.40-1.36(m，3H)。MS：[M+H]+＝567.7。

example 8,

Preparation of (SZY1805-1k) Compound

The above compounds were prepared according to the synthetic scheme shown in figure 8:

SZY-1805-6(10g) is dissolved in 50ml DCM, DIPEA (6g, 31.47mmol, 3eq, MW129.25) is added into the DCM, stirring reaction is carried out, hexanoyl chloride (5.0g) is added in batches at 0 ℃, the hexanoyl chloride is added in half an hour or so, after the end of low price, the reaction is stirred overnight after the room temperature is raised to room temperature, the reaction is monitored to be finished by T L C on the next day, the reaction solution is washed for 2 times by 100ml of water, and is washed for 2 times by 100ml of saturated saline solution, an organic phase is dried by anhydrous sodium sulfate, rotary evaporation and column chromatography separation are carried out, so 8.5g of oily liquid is obtained, and the yield is 81% (SZY1805-1 k).

The structure identification data is as follows:

1H NMR:SZY1805-1k(400MHz CDCl3)5.19(s，2H)，5.08-5.05(t，1H)，4.52-4.49(m，2H)，3.80(s，3H)，3.23-3.12(m，4H)，3.12-3.11(m，4H)，2.74-2.68(m，2H)，2.46-2.40(m，2H)，2.30-2.26(m，2H)，2.24(s，3H)，1.77(s，3H)，1.46-1.44(m，6H)，1.56-1.36(m，6H)，0.96-0.94(t，3H)。MS：[M+H]+＝518.6。

examples 9,

Preparation of (SZY1805-1M) Compound

The above compounds were prepared according to the synthetic scheme shown in figure 9:

SZY-1805-6(10g) was dissolved in 50ml DCM, DIPEA (6g, 31.47mmol, 3eq, MW129.25) was added to it, the reaction was stirred, succinic anhydride (5.0g) was added at 0 ℃ and the reaction was stirred overnight with warming to room temperature, the reaction was monitored for completion the next day by T L C, the reaction solution was washed 2 times with 100ml of water, 2 times with 100ml of saturated brine, the organic phase was dried over anhydrous sodium sulfate, rotary evaporated, and separated by column chromatography to give 7.0g of oily liquid in 68% yield (SZY-1805-1M).

The structure identification data is as follows:

1H NMR:SZY1805-1M(400MHz CDCl3)5.16(s，2H)，5.08-5.05(t，1H)，4.52-4.49(m，2H)，3.80(s，3H)，3.23-3.12(m，4H)，3.12-3.11(m，4H)，2.74-2.68(m，2H)，2.46-2.40(m，2H)，2.30-2.26(m，4H)，2.24(s，3H)，1.77(s，3H)，1.46-1.44(m，3H)，1.40-1.36(m，3H)。MS：[M+H]+＝520.5。

examples 10,

Preparation of (SZY 1805-1L) Compound

SZY-1805-6H (5.33g, 11.5mmol, 1.0eq, MW463.52) is dissolved in 50ml DCM, DIPEA (4.46g, 34.5mmol, 3eq, MW129.25) is added to the DCM, stirring is carried out for reaction, acetyl chloride (1.50g, 17.5mmol, 1.5eq, MW92.52) is added in portions at 0 ℃, the addition is completed by about half an hour, after the completion of the reaction, the reaction is stirred overnight at room temperature, T L C monitors the completion of the reaction on the next day, the reaction liquid is washed 2 times with 100ml of water, the saturated saline water is washed 2 times with 100ml, the organic phase is dried with anhydrous sodium sulfate, rotary evaporation is carried out, column chromatography is carried out, and 5.00g of oily liquid is obtained, and the yield is 80% (SZY 1805-7L).

And (2) dissolving SZY-1805-7L (5g) in 20ml of EA, adding 4M HCl 50ml of EA, stirring at normal temperature for reaction, monitoring every half hour by T L C, wherein the reaction is completed within 1.5h-2h and no obvious impurity is shown, removing the solvent by rotary evaporation after the reaction is completed to obtain an oily substance, adding a sodium bicarbonate aqueous solution to neutralize hydrochloric acid, extracting with ethyl acetate, drying with anhydrous sodium sulfate, and evaporating the solvent to obtain 2.8g of a product (SZY 1805-1L).

The structure identification data is as follows:

1H NMR:SZY1805-1L(400MHz CDCl3)8.25(s，2H)，5.17(s，2H)，5.09-5.04(t，1H)，4.35(s，2H)，3.80(s，3H)，3.33-3.21(m，4H)，2.75-2.69(m，2H)，2.50-2.46(m，2H)，2.20-2.16(m，2H)，2.10(s，3H)，1.76(s，3H)。MS：[M+H]+＝406.4

examples 11,

Preparation of (SZY1805-1p) Compound

The compound SZY1805-1p HCl was prepared according to the scheme shown in FIG. 10.

The specific operation is as follows:

1: SZY1805-3p Synthesis

To a 1000ml round bottom flask was added SZY1805-2p (100.0g, 312.2mmol, 1.0eq), dissolved by 600ml DMF, added imidazole (95.65g, 1.45mmol, 4.5eq) and stirred for 0.5h, then added TBSCl (117.61g, 780.4mmol, 2.5eq) in portions and stirred for reaction at room temperature overnight.

The reaction was followed by detection of T L C (n-heptane: ethyl acetate: 1:2, Rf)₁＝0.8，Rf₂0.3), adding 3.0L water to the solution, adding 300ml 4 of EA to extract, combining and concentrating the organic phases, adding 3.0L water to the organic phases, adding 300ml 4 of EA to extract, combining and concentrating the organic phases, adding 3.0L water to the organic phases, adding 300ml 4 of EA to extract, combining and removing most of the organic phases by rotary evaporation and then directly carrying out the next step.

2: SZY1805-4p Synthesis

Adding 400ml of THF, 400ml of water and 400ml of acetic acid into the product obtained in the last step, stirring and reacting for about 2 hours, detecting that the reaction is finished by T L C (n-heptane: ethyl acetate: 1:2, Rf: 0.3), adding 3L water into the reaction liquid, adding DCM400ml to extract, merging and concentrating organic phases, adding 3L water into the organic phase, adding DCM400ml to extract, merging and concentrating the organic phases, adding 3L water into the organic phase, adding DCM400ml to extract, merging and spin-drying the organic phases to obtain white solid powder, washing the white solid powder with 500ml of n-heptane, carrying out suction filtration and drying to obtain 129g of white solid powder with the yield of 95%.

3: SZY1805-5p Synthesis

SZY1805-4p (36.82g, 84.72mmol, 1.0eq) was dissolved in 360ml dcm, Boc aminopropanol, EDCI, DMAP were added to the reaction flask in sequence, and the reaction was stirred for 5h, followed by T L C detection of the completion of the reaction (n-heptane: ethyl acetate 1:2, Rf 0.8), and the reaction was rotary evaporated to remove part of the organic solvent, and then directly subjected to the next reaction.

4: SZY1805-6p Synthesis

To the reaction in the previous step was added 500ml THF, TBAF.3H₂O(29.4g，93.19mmol1.1eq), stirring for 1h, detecting by T L C that the reaction is complete, adding 400ml of 1% aqueous HCl to the reaction mixture, extracting with 100ml of 4 DCM, combining the organic phases, washing the organic phase with 400ml of 1% aqueous HCl, extracting the aqueous phase with 100ml of 4 DCM, combining the concentrated organic phases, washing the organic phase with 400ml of 1% aqueous HCl, extracting the aqueous phase with 100ml of 4 DCM, combining the dried organic phases to give about 40.46g of a colorless oil, with a 100% yield.

5: SZY1805-7p Synthesis

SZY1805-6p (40.46g, 84.72mmol, 1.0eq) is dissolved in 400ml DCM, DIPEA (32.85g, 254.16mmol, 3.0eq) is added under ice-water bath conditions, stirring reaction is carried out for 0.5h, propionyl chloride (11.76g, 127.08mmol, 1.5eq) is added in portions, stirring reaction is carried out overnight, the reaction is detected to be finished by T L C the next day, 400ml water is added into the reaction solution, 100ml DCM is used for 4 times of extraction, the organic phase is washed by 400ml water, the aqueous phase is extracted by DCM100ml times 4 times, the organic phase is concentrated, the organic phase is successively washed by 400ml water, 100ml DCM is used for 100ml 4 times of extraction, combined spin drying is carried out to obtain about 45g of red brown oil, the yield is 96% (HP L C monitoring, if the purity is lower, column chromatography purification is carried out)

6: SZY1805-1p HCl Synthesis

Dissolving SZY1805-7p (45g) in 400ml 2 mol/L EA. HCl, stirring at normal temperature for reaction for 1h, precipitating a large amount of white solid powder, filtering, collecting a filter cake, washing the filter cake with 200ml EA slurry for 1h, performing suction filtration to collect the filter cake, performing reverse EA slurry washing for three times, filtering and drying to obtain 25g of white solid powder, wherein the yield is 71.4%, and the purity of HP L C is 98.9%.

The structure identification data is as follows:

1H NMR:SZY1805-1 p(400MHz CDCl3)8.29(s，2H)，5.16(s，2H)，5.08-5.05(t，1H)，4.13(s，2H)，3.79(s，3H)，3.34-3.33(d，J＝6.8Hz,2H)，3.06(s,2H),2.74-2.68(m，2H)，2.41-2.37(m，2H)，2.28-2.24(m，2H)，2.22(s，3H)，2.08(s,2H),1.77(s，3H),1.30-1.26(t,3H)。MS：[M+H]⁺＝433.7.

examples 12,

Preparation of (SZY1805-1q) CompoundPrepare for

The above compounds were prepared according to the synthetic scheme shown in figure 11:

the specific operation is as follows:

1: SZY1805-3q Synthesis

To a 1000ml round bottom flask was added SZY1805-2q (100.0g, 312.2mmol, 1.0eq), dissolved by 600ml DMF, added imidazole (95.65g, 1.45mmol, 4.5eq) and stirred for 0.5h, then added TBSCl (117.61g, 780.4mmol, 2.5eq) in portions and stirred for reaction at room temperature overnight.

2: SZY1805-4q Synthesis

Adding 400ml of THF, 400ml of water and 400ml of acetic acid into the product obtained in the last step, stirring and reacting for about 2 hours, detecting that the reaction is finished by T L C (n-heptane: ethyl acetate: 1:2, Rf: 0.3), adding 3L water into the reaction liquid, adding DCM400ml to extract, merging and concentrating organic phases, adding 3L water into the organic phase, adding DCM400ml to extract, merging and concentrating the organic phases, adding 3L water into the organic phase, adding DCM400ml to extract, merging and spin-drying the organic phases to obtain white solid powder, washing the white solid powder with 500ml of n-heptane, carrying out suction filtration and drying to obtain 125g of white solid powder with the yield of 97%.

3: SZY1805-5q Synthesis

SZY1805-4q (36.82g, 84.72mmol, 1.0eq) was dissolved in 360ml dcm, Boc aminoethylamine, EDCI, DMAP were added to the reaction flask in sequence, and the reaction was stirred for 5h, followed by T L C detection of the completion of the reaction (n-heptane: ethyl acetate 1:2, Rf 0.8), and the reaction was rotary evaporated to remove part of the organic solvent, and then the next reaction was carried out directly.

4: SZY1805-6q Synthesis

To the reaction in the previous step was added 500ml THF, TBAF.3H₂O(294g, 93.19mmol, 1.1eq), stirring for 1h, detecting completion of the reaction by T L C, adding 400ml of 1% aqueous HCl to the reaction mixture, extracting with 100ml of 4 DCM, combining the organic phases, washing the organic phase with 400ml of 1% aqueous HCl, extracting the aqueous phase with 100ml of 4 DCM, combining the concentrated organic phases, washing the organic phase with 400ml of 1% aqueous HCl, extracting the aqueous phase with 100ml of 4 DCM, combining the dried organic phases to give about 39.02g of a colorless oil with 98% yield.

5: SZY1805-7q Synthesis

SZY1805-6q (39.02g, 84.72mmol, 1.0eq) was dissolved in 400ml DCM, DIPEA (32.85g, 254.16mmol, 3.0eq) was added under ice-water bath conditions and stirred for 0.5h, then propionyl chloride (11.76g, 127.08mmol, 1.5eq) was added in portions and stirred overnight, the reaction was checked for completion by T L C the next day, 400ml water was added to the reaction solution, 100ml DCM was used for 4 extractions, the organic phase was combined, 400ml water was used for washing the aqueous phase, 100ml DCM was used for 4 extractions, the organic phase was combined with DCM, 400ml water was used for washing the organic phase successively, 100ml water was used for extraction, 100ml water was used for 4 extractions, combined with spin-drying to give a red-brown oil, yield was 92% (. HP L C monitoring, column chromatography purification if purity is lower)

6: SZY1805-1q HCl Synthesis

Dissolving SZY1805-7q (42g) in 400ml 2 mol/L EA. HCl, stirring at normal temperature for reaction for 1h, precipitating a large amount of white solid powder, filtering, collecting filter cake, washing the filter cake with 200ml EA slurry for 1h, filtering, collecting the filter cake, washing with EA slurry for three times, filtering and drying to obtain 28g of white solid powder, wherein the yield is 73%, and the purity of HP L C is 99.2%.

1H NMR:SZY1805-1q(400MHz CDCl3)8.28(s，2H)，5.18(s，2H)，5.08-5.04(t，1H)，4.15(s，2H)，3.81(s，3H)，3.34-3.31(d，J＝6.8Hz,2H)，3.08(s,2H),2.74-2.66(m，2H)，2.43-2.37(m，2H)，2.26-2.22(m，2H)，2.18(s，3H)，2.05(s,2H),1.78(s，3H),1.30-1.26(t,3H)。MS：[M+H]⁺＝419.5。

The related hydrochloride of each compound can be prepared in ethyl acetate hydrochloride, and the related tartrate can be prepared by adding tartaric acid into acetone solution.

Example 13 stability test

Tartrate salt of the compound prepared in example 3, having the formula

Stability test: SZY1805-1f-JSS stability test statistical table under different solutions

Solutions of	0h(％)	24h(％)	72h(％)	5d(％)	Remarks for note
						Deionized water	99.25	99.15	98.78	98.72
Physiological saline	99.25	99.17	98.85	98.49
						pH6.3 phosphate buffer	97.76	92.17	76.62
pH6.5 phosphate buffer	98.85	96.37	90.33		With an overlap
						pH6.8 phosphate buffer	93.98	77.17	42.90
pH7.0 phosphate buffer	90.97	68.93	28.96

Note that the purity is the relative purity of the liquid phase (HP L C), and the product peak at 254nm, pH6.5 has an overlap with the impurity peak, and the non-separation results in a wrong trend.

The hydrochloride salt of the compound prepared in example 1, having the formula:

SZY1805-1H stability test statistical table under different solutions:

solutions of	0h(％)	24h(％)	72h(％)	5d(％)	Remarks for note
						Deionized water	98.99	98.70	97.67	97.06
Physiological saline	99.01	98.92	98.70	98.05
						pH6.3 phosphate buffer	97.77	86.38	60.80
pH6.5 phosphate buffer	93.89	74.21	44.13
						pH6.8 phosphate buffer	92.17	59.71	19.52
pH7.0 phosphate buffer	93.79	49.60	8.81

Note that the purity is the relative purity of the liquid phase (HP L C), and the wavelength is 254 nm.

Liquid phase conditions:

phase A: 0.05% phosphoric acid aqueous solution

Phase B: acetonitrile

Time (min)	A％	B％
			0	90	10
20	10	90
			25	10	90
26	90	10
			35	90	10

Column temperature: 30 deg.C

Flow rate: 1ml/min

Sample size 10 mu L

Sample concentration 1mg/m L

Detection wavelength: 254nm

A chromatographic column: agilent Zorbax SB C18

The stability of the compounds prepared in the examples in physiological saline solution is shown in the following table (PH 6, storage temperature 25 ℃): (the following compounds except for mycophenolate sodium are hydrochloride salts of corresponding compounds)

Compound (I)	0h(％)	24h(％)	72h(％)	5d(％)
					SZY1805-1a	99.01	99.00	98.99	98.85
SZY1805-1b	98.86	98.20	97.99	97.32
					SZY1805-1c	98.99	97.11	96.99	95.69
SZY1805-1d	99.23	99.16	98.99	98.87
					SZY1805-1e	99.56	97.02	96.99	96.55
SZY1805-1k	99.21	99.00	98.79	98.65
					SZY1805-1M	98.97	98.85	98.55	98.46
SZY1805-1p	99.02	99.01	98.86	98.78
					SZY1805-1q	99.56	99.53	99.49	99.40
Mycophenolate sodium	98.97	93.00	94.32	91.33
					SZY1805-1H	99.02	99.01	98.86	98.78
SZY1805-1f	99.56	99.53	99.49	99.40
					SZY1805-1L	98.97	98.50	98.32	98.33

(the following table shows the stability test results of sodium mycophenolate, SZY1805-1f, SZY 1805-1L in water:

stability Studies (Water) of sodium mycophenolate (about 0.25mg in 1 ml) solution

Stability test data for SZY1805-1f (hydrochloride): (approximately 0.25mg in 1 ml) stability study (Water)

Stability test data of SZY 1805-1L (about 0.25mg in 1 ml) stability test (Water)

The compounds in the general formula have different physicochemical properties according to different structures, and the stability in water, physiological saline and buffer salt is different, so that the compounds in the general formula are stable on the whole.

Example 14 pharmacokinetic experiments

Test article mycophenolate sodium (mycophenolate sodium): white powdery solid, molecular weight: 342.32, batch number: BSP-20171123, purity: 99.0%, preservation conditions: sealing and storing at normal temperature, and the validity period is 2019.11.22.

Test article SZY1805-1f JSS: light yellow viscous liquid, molecular weight: 625.66, purity: is greater than 98 percent.

Test article SZY1805-1H HCl: white solid, molecular weight: 455.93, purity: 98.8 percent.

Detection compound: mycophenolate sodium

Eye drops are applied to eyes, and each time of application helps the eyes to slightly close the double eyelids for about 10 seconds.

Administration vehicle: sodium chloride injection

Equimolar dosing: 0.16 μ M/new zealand rabbit/time; that is, the mycophenolate sodium group is administered with mycophenolate sodium 0.055 mg/New Zealand rabbit, the SZY1805-1f group is selected to be administered with mycophenolate sodium 0.1 mg/New Zealand rabbit, the SZY1805-1H group is selected to be administered with mycophenolate sodium 0.073 mg/New Zealand rabbit

100u L/New Zealand Rabbit

SZY1805-1f JSS is 1mg/m L1805-1H HCl 0.73mg/m L, mycophenolate sodium 0.55mg/m L

The lower limit of the detection quantification is 10ng/m L

As a result: 1) after SZY1805-1f JSS and SZY1805-1H HCl are respectively administrated to eyes in equimolar (0.16 mu M/new Zealand rabbit dose), the active metabolite mycophenolate sodium can be detected in the new Zealand rabbit aqueous humor; 2) in the rabbit aqueous humor after 1H of administration, the contents of mycophenolate sodium in groups SZY1805-1f and SZY1805-1H are close, namely 724.4 +/-313.6 ng/ml and 730.6 +/-434.0 ng/ml, which are respectively higher than 180.0 +/-72.2 ng/ml of mycophenolate sodium; 3) no irritation was produced to New Zealand rabbit eyes at this SZY1805-1f JSS, SZY1805-1H HCl dosing concentration.

New Zealand rabbit eye part was given 0.16. mu.M/mouse of SZY1805-1f JSS, SZY1805-1H HCl, and mycophenolate sodium for 1H, followed by mycophenolate sodium concentration (ng/ml) in aqueous humor

The following compounds were subjected to local pharmacokinetic experiments in the manner described above, and the results were as follows:

concentration of mycophenolate sodium in aqueous humor (ng/ml) after 0.16. mu.M/rabbit eye administration of New Zealand eyes

Note: all the above compounds are corresponding hydrochloride

EXAMPLE 15 eye drops of the Compounds of the examples Rabbit eye irritation test

The purpose is as follows: the eye drops prepared from the compounds of the above examples in different concentrations were observed to produce irritation to rabbit eyes.

The method comprises the following steps: 165 male New Zealand rabbits with no secretion in both eyes, no redness of conjunctiva, no abnormality of cornea and iris were divided into 55 groups and 3 rabbits/group, and 0.025%, 0.05%, 0.1%, 0.2%, 0.4% of the example compound (SZY1805-1a, SZY1805-1b, SZY1805-1c, SZY1805-1d, SZY1805-1e, SZY1805-1k, SZY1805-1M, SZY1805-1f, SZY1805-1H, SZY1805-1p, SZY1805-1q) eye drops were administered. Using the same-body left-right-side self-contrast method, the eye drops of the compound of the example at the corresponding concentration were administered to the left eye of each group of animals, and the same amount of physiological saline, 50. mu.l/eye, was administered to the right eye. D1 was administered 1 time followed 3 times daily (about 4 hours apart) for 14 consecutive days (d1-d 14). Observing the eye irritation condition of the rabbits at 1, 2, 4 and 24 hours after the first administration, 1, 2 hours after each administration and 1, 2, 4, 24, 48 and 72 hours after the last administration, recording the score, and finally scoring according to the eye irritation evaluation standard. At the end of the test, 3 animal eyeballs and other abnormal animal eyeballs in the eye drop group of the compound of the example with the concentration of 0.4% are picked up for histopathological examination.

As a result: the administration is carried out for 14 days continuously, and the visual observation result shows that the secretion of rabbit eyes at the administration side of each group is increased compared with that at the control side during the administration period, conjunctiva has no hyperemia and edema, cornea is transparent and has no turbidity, iris texture is clear, and the rabbit eyes have no hyperemia and swelling, and the eye irritation score of all animals is 0-2. Each group of each observation point was evaluated for irritability of less than 3 points, and was non-irritating (evaluation criteria for eye irritability: 0-3 points: no irritability).

And (4) conclusion: the eye drops of the compound of the embodiment with the concentration of 0.025%, 0.05%, 0.1%, 0.2% and 0.4% have no obvious irritation to normal rabbit eyes after being continuously administrated for 14 days.

Note: the animal experiment was not completed and 14 days of dosing was complete and in the observation period. In view of detailed observation, after 72 hours of observation after the last administration, 3 animal eyeballs and other abnormal animal eyeballs of the eye drop group of the compound of the example with the concentration of 0.4% were picked up and sent to the pathology for histopathological examination.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, ester, solvate or isomer thereof:

in the formula I, R₁、R₂Independently selected from: -NH₂H, amino/hydroxy/halogen/aryl/carboxy-substituted C₁-C₁₀Alkyl, amino/hydroxy/halogen substituted C₃-C₆Cycloalkyl radical, C₁-C₁₀Alkyl radical, C₃-C₆Cycloalkyl, -N (CH)₂CH₃)₂、-N(CH₃)₂Any one of the above;

R₃is H or-CH₃，

R₄、R₅Independently selected from: -H, C₁-C₅Any one of alkyl groups;

x represents O or N;

n is an integer of 1 to 8.

2. The compound of claim 1, wherein: the compound shown in the formula I is any one of the following compounds:

3. a process for the preparation of a compound according to claim 1 or 2, comprising the steps of:

in the formula II R₄、R₅X is as defined in claim 1 wherein R is₄、R₅X, n is an integer of 1 to 8;

in the formula III, R₁、R₂、R₃The same claim as in claim 1, wherein R₁、R₂、R₃The definition of (1).

4. The method of claim 3, wherein: the condensation reaction is carried out in the presence of an acid-binding agent;

the acid-binding agent is at least one of DIPEA, triethylamine and pyridine;

the mol ratio of the compound shown in the formula II to the compound shown in the formula III and the acid-binding agent is as follows: 1:1-2: 2-4;

the reaction temperature is room temperature, and the reaction time is 8-14 h.

5. Use of a compound of formula I as claimed in claim 1 or a pharmaceutically acceptable salt, ester, solvate or isomer thereof as a prodrug of mycophenolic acid.

6. Use of a compound of formula I as defined in claim 1, or a pharmaceutically acceptable salt, ester, solvate or isomer thereof, for the preparation of an immunosuppressant; in particular, the application is the application in the preparation of medicines for preventing or treating transplant rejection and immune-mediated and/or inflammatory diseases.

7. A pharmaceutical formulation comprising a compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt, ester, solvate or isomer thereof, and one or more of: excipient, diluent, pH stabilizer.

8. The pharmaceutical formulation of claim 7, wherein: the pharmaceutical preparation can be prepared into various dosage forms including liquid preparations; specifically, the liquid preparation is at least one of injection, oral solution and eye liquid preparation.

9. The pharmaceutical formulation of claim 8, wherein: the liquid preparation is not added with a cosolvent, and specifically the cosolvent is polysorbate 80.

10. An ophthalmic liquid preparation comprising a compound of formula I as shown in claim 1 or a pharmaceutically acceptable salt, ester, solvate or isomer thereof, a solvent, one or more physiologically acceptable excipients; specifically, the solvent is water; more specifically, the ophthalmic liquid formulation does not add a co-solvent.