CN111393399B - Prodrugs of mycophenolic acid and method for preparing same - Google Patents

Prodrugs of mycophenolic acid and method for preparing same Download PDF

Info

Publication number
CN111393399B
CN111393399B CN202010419146.5A CN202010419146A CN111393399B CN 111393399 B CN111393399 B CN 111393399B CN 202010419146 A CN202010419146 A CN 202010419146A CN 111393399 B CN111393399 B CN 111393399B
Authority
CN
China
Prior art keywords
added
reaction
szy1805
water
szy
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010419146.5A
Other languages
Chinese (zh)
Other versions
CN111393399A (en
Inventor
王延东
刘国强
刘伟
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Guangzhou Ocusun Ophthalmic Biotechnology Co Ltd
Original Assignee
Guangzhou Ocusun Ophthalmic Biotechnology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Guangzhou Ocusun Ophthalmic Biotechnology Co Ltd filed Critical Guangzhou Ocusun Ophthalmic Biotechnology Co Ltd
Publication of CN111393399A publication Critical patent/CN111393399A/en
Application granted granted Critical
Publication of CN111393399B publication Critical patent/CN111393399B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
    • C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D307/87—Benzo [c] furans; Hydrogenated benzo [c] furans
    • C07D307/88—Benzo [c] furans; Hydrogenated benzo [c] furans with one oxygen atom directly attached in position 1 or 3
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0048—Eye, e.g. artificial tears
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection

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 in formula I: compared with mycophenolic acid, the compound shown in the formula I has better aqueous permeability, better aqueous solution stability and better water solubility, and can be hydrolyzed into mycophenolic acid ester in aqueous humor, so that the drug concentration of the mycophenolic acid in aqueous humor is obviously improved compared with that of mycophenolate sodium, and the concentration of the mycophenolic acid in aqueous humor is 2 times or higher than that of the mycophenolate sodium in experiments, and the compound is more suitable for preparing eye liquid preparations.

Description

Prodrugs of mycophenolic acid and method for preparing same
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 (Mycophenolate mofetil, MMF) is a potent, novel immunosuppressant that is a 2-morpholinoethyl product of mycophenolic acid (mycophenolic acid, mycophenolic acid, MPA). Inosine monophosphate dehydrogenase is an enzyme required for the synthesis of guanosine by lymphocytes and is thus necessary for DNA synthesis and cell proliferation. MMF is a non-competitive, reversible inhibitor of this enzyme. Inhibition of hypoxanthine monophosphate dehydrogenase by MPA can cause cells to stay 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 effects on lymphocyte proliferation, MMF can inhibit glycosylation of glycoproteins in lymphocytes and monocytes that are involved in adhesion to endothelial cells. The mycophenolate sodium enteric coated tablet is suitable for being used together with cyclosporin and corticosteroid, and is used for preventing acute rejection of adult patients receiving allogeneic kidney transplantation.
The existing commercially available forms of mycophenolate mofetil, which are clinically used at present, comprise tablets and injections, and polysorbate 80 is required to be added into the injections as a cosolvent due to poor solubility. The commercially available forms of sodium mycofenate are capsules and tablets. The aqueous solutions of mycophenolate mofetil and sodium mycophenolate are both unstable and hydrolyze to mycophenolic acid in aqueous solution, and the phenolic hydroxyl groups are easily oxidized to unknown impurities. The above factors make it impossible to directly prepare mycophenolate mofetil and sodium mycophenolate into liquid preparations (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:
Figure BDA0002496194080000021
in the above formula I, R 1 、R 2 Can be independently selected from: -NH 2 -H, amino/hydroxy/halogen/aryl/carboxyl substituted C 1 -C 10 Alkyl (specifically, amino/hydroxy/halogen/aryl/carboxyl substituted C 1 -C 5 Alkyl radicals, e.g. C 6 H 5 CH 2 -、-CH 2 CH 2 COOH), amino/hydroxy/halogen substituted C 3 -C 6 Cycloalkyl, C 1 -C 10 Alkyl (which may be C in particular) 1 -C 5 Alkyl radicals, e.g. -CH 2 CH 2 CH 2 CH 3 、-CH 3 )、C 3 -C 6 Cycloalkyl, -N (CH) 2 CH 3 ) 2 、-N(CH 3 ) 2 Any one of them;
R 3 is H or-CH 3 ，
R 3 When H is the same, R 1 、R 2 Not both-H; r is R 3 When H is the same, R 1 、R 2 Not being one of H and the other of-NH 2 ；
R 4 、R 5 Can be independently selected from: h, C A-5, 1 -C 5 alkyl (which may be C in particular) 1 -C 3 Alkyl radicals, e.g. -CH 3 、-CH 2 CH 3 ) Any one of them;
x represents O or N;
n is an integer of 1 to 8, specifically 1 to 5 or 1 to 3, and more specifically 1 or 2.
In some of these embodiments, the compounds of the present invention may be exemplified by, but are not limited to, the structures shown below:
Figure BDA0002496194080000022
/>
Figure BDA0002496194080000031
/>
Figure BDA0002496194080000041
the compound shown in the formula I is prepared by a method comprising the following steps:
enabling the compound shown in the formula II and the compound shown in the formula III to undergo condensation reaction to obtain the compound shown in the formula I;
Figure BDA0002496194080000042
r in formula II 4 、R 5 X is as in formula I 4 、R 5 X is defined, n is an integer from 1 to 8, and can be specifically 1;
r in formula III 1 、R 2 、R 3 R in the same manner as in I 1 、R 2 、R 3 Is defined as follows;
in the above method, the condensation reaction is performed in the presence of an acid-binding agent;
the acid binding agent can be DIPEA, triethylamine, pyridine and the like, and can be specifically DIPEA;
the molar ratio of the compound shown in the formula II to the compound shown in the formula III and the acid binding agent can be as follows: 1:1-2:2-4, and can be specifically 1:1.5:3;
the condensation reaction is carried out in an organic solvent, which may specifically be DCM;
the temperature of the reaction may be room temperature and the time may be 8-14 hours.
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 thereof (including stereoisomers, enantiomers, tautomers or mixtures thereof).
The salt may be in particular hydrochloride or tartaric acid, etc.
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 thereof (including stereoisomers, enantiomers, tautomers or mixtures thereof) as a prodrug of mycophenolic acid;
the use of a compound of formula I or a pharmaceutically acceptable salt, ester, solvate or isomer thereof (including stereoisomers, enantiomers, tautomers or mixtures thereof) in the preparation of an immunosuppressant;
specifically, the application is the application of a compound shown in the formula I or pharmaceutically acceptable salt, ester, solvate or isomer (including stereoisomer, enantiomer, tautomer or mixture thereof) thereof in preparing medicines for preventing or treating transplant rejection and immune-mediated and/or inflammatory diseases.
The present 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 the following: excipient, diluent and pH stabilizer.
The pharmaceutical formulation can be formulated into various dosage forms including liquid formulations.
The liquid preparation can be at least one of injection, oral solution and eye liquid preparation.
The liquid formulation was free of co-solvents including polysorbate 80.
The invention also provides an eye liquid preparation, which comprises a compound shown in a formula I or pharmaceutically acceptable salt, ester, solvate or isomer thereof, a solvent and one or more physiologically acceptable auxiliary materials.
In the eye liquid preparation, the solvent is water.
The ocular liquid formulation is free of the addition of a co-solvent including polysorbate 80.
Compared with mycophenolic acid, mycophenolate sodium and mycophenolate ester, the compound shown in the general formula has the following advantages: 1. has better water solubility; 2. the mycophenolate mofetil is more stable than the mycophenolate sodium and the mycophenolate mofetil in a system containing water components, and almost has no degradation; 3. the medical composition has better permeability, can penetrate cornea, enter aqueous humor, has higher aqueous humor drug concentration which is 2-5 times of mycophenolic acid; 4. the aqueous humor can be hydrolyzed into mycophenolic acid to exert the drug effect.
Compared with mycophenolic acid, the compound shown in the formula I has better aqueous permeability, so that the drug concentration of the mycophenolic acid in aqueous humor is obviously improved compared with that of mycophenolic acid sodium, and experiments show that the drug concentration of the compound in the examples 1-11 in the aqueous humor of New Zealand rabbit after administration is 2 times or more than that of the mycophenolic acid sodium, and the compound is more suitable for preparing eye liquid preparations.
Drawings
FIG. 1 is a flow chart showing the preparation of the objective compound in example 1 of the present invention.
FIG. 2 is a flow chart showing the preparation of the objective compound in example 2 of the present invention.
FIG. 3 is a flow chart showing the preparation of the objective compound in example 3 of the present invention.
FIG. 4 is a flow chart of the preparation of the target compound in example 4 of the present invention.
FIG. 5 is a flow chart showing the preparation of the objective compound in example 5 of the present invention.
FIG. 6 is a flow chart showing the preparation of the objective compound in example 6 of the present invention.
FIG. 7 is a flow chart showing the preparation of the target compound in example 7 of the present invention.
FIG. 8 is a flow chart showing the preparation of the objective compound in example 8 of the present invention.
FIG. 9 is a flow chart showing the preparation of the objective compound in example 9 of the present invention.
FIG. 10 is a flow chart showing the preparation of the objective compound in example 11 of the present invention.
FIG. 11 is a flow chart showing the preparation of the objective compound in example 12 of the present invention.
Detailed Description
The present invention will be described with reference to the following 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; the reagents, materials, etc. used in the examples described below are commercially available unless otherwise specified.
For preparing a stable ophthalmic liquid preparation from a mycophenolate sodium or a mycophenolate mofetil derivative, three conditions are required: 1. can be decomposed into mycophenolic acid in aqueous humor; 2. can be kept stable in aqueous solutions; 3. has good water solubility, and can be prepared into aqueous solution preparation.
The mycophenolic acid sodium is designed into a series of compounds, and through an eye pharmacokinetics experiment and a stability experiment, the compounds with the following general formula can meet the requirements, and can be prepared into an eye liquid preparation to be used as a prodrug of the mycophenolic acid so as to locally exert the drug effect of the mycophenolic acid. R is R 1 、R 2 、R 3 The choice of (c) plays a critical role in whether or not it is stable and whether it is capable of hydrolysing to mycophenolic acid in aqueous humor, as observed in experiments, when R 1 Is H, R 2 Is H, R 3 In the case of H, the compounds of formula I are not stable in aqueous solution and are subject to hydrolysis, but waterThe decomposed phenolic hydroxyl groups generate oxidation reaction, R 1 Is H, R 2 Is NH 2 ,R 3 In the case of H, the same applies. The compound shown in the formula I has better aqueous permeability than the mycophenolic acid, so that the drug concentration of the mycophenolic acid in aqueous humor is obviously improved compared with that of the mycofenac sodium, 3 times or more of the mycophenolic acid sodium is observed in experiments, the preparation method is more suitable for preparing eye liquid preparations,
Figure BDA0002496194080000061
in the above formula I, R 1 、R 2 Can be independently selected from: -NH 2 -H, amino/hydroxy/halogen/aryl/carboxy substituted C 1 -C 10 Alkyl (specifically, amino/hydroxy/halogen/aryl/carboxyl substituted C 1 -C 5 Alkyl radicals, e.g. C 6 H 5 CH 2 -、-CH 2 CH 2 COOH), amino/hydroxy/halogen substituted C 3 -C 6 Cycloalkyl, C 1 -C 10 Alkyl (which may be C in particular) 1 -C 5 Alkyl radicals, e.g. -CH 2 CH 2 CH 2 CH 3 、-CH 3 )、C 3 -C 6 Cycloalkyl, -N (CH) 2 CH 3 ) 2 、-N(CH 3 ) 2 Any one of them;
R 3 is H or-CH 3 ，
R 3 When H is the same, R 1 、R 2 Not both-H; r is R 3 When H is the same, R 1 、R 2 Not being one of H and the other of-NH 2 ；
R 4 、R 5 Can be independently selected from: h, C A-5, 1 -C 5 alkyl (which may be C in particular) 1 -C 3 Alkyl radicals, e.g. -CH 3 、-CH 2 CH 3 ) Any one of them;
x represents O or N;
n is an integer of 1 to 8, and may be specifically 1.
Example 1,
Figure BDA0002496194080000071
Preparation of (SZY 1805-1H) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 1:
to a 250ml round bottom flask was added SZY-1805-2H (10.0 g,31.22mmol,1.0eq, MW320.34), 100ml DMF was added to dissolve, imidazole (13.8 g,202.9mmol,6.5eq, MW68.08) was added to dissolve, then TBSCl (18.82 g,124.87mmol,4.0eq, MW150.7) was added in portions and the reaction was stirred at room temperature overnight. After completion of the TLC detection reaction, 150ml of EA and 200ml of water were added to the solution for delamination. The aqueous phase was extracted three times with 50ml ea and the organic phases were combined. The organic phase was washed with 1% hydrochloric acid solution and 3-5 times with pure water (removal of DMF). The organic phase was dried over anhydrous sodium sulfate and distilled off to give an oil. To the oil was added 50ml of THF, 50ml of water, 50ml of acetic acid, and the reaction was stirred at room temperature and monitored by TLC after 2h. To the reaction solution was added 150ml of EA and 200ml of water were separated, the aqueous phase was extracted 3 times with 50ml of EA, and the organic phases were combined. After washing the organic phase with water 5 times, it was dried over anhydrous sodium sulfate and distilled off to give a white solid (containing a small amount of solvent). Adding 50ml of PE, pulping, and suction filtering to obtain 12.0g of white solid with a yield of 88.4% (SZY-1805-4H);
to a 250ml round bottom flask was added SZY-1805-4H (5.0 g,11.5mol,1.0eq, MW434.6) and dissolved by adding 50ml dichloromethane. Sequentially adding BocNHC 2 H 4 OH(2.22g，13.8mmol，1.2eq，
MW 161.2), DMAP (150 mg,1.2mmol,0.1eq, MW122), EDCI (2.6 g,13.8mmol,1.2eq, MW191.7), and stirring the reaction at room temperature for 7h. After the completion of the TLC, DCM was removed by rotary evaporation under reduced pressure, and 150ml of EA was added and 150ml of water was separated. The organic phase was washed with 100ml of water 2 times, with 100ml of saturated brine 2 times, dried over anhydrous sodium sulfate, and filtered and distilled to give 6.05g of an oil with a yield of 91% (SZY-1805-5H).
To a 250ml round bottom flask containing SZY-1805-5H (6.05 g,10.47mmol,1.0eq, MW577.8) was added 50ml THF to dissolve, then TBAF (3.96 g,12.56mmol,1.2eq, MW315.5) was added and the reaction was stirred at room temperature. The solution became wine red after the addition of TBAF and then orange yellow. After 3h, TLC was monitored and the reaction was complete. To this was added 100ml of water and EA, and the layers were separated. The organic phase was washed with water, dried over anhydrous sodium sulfate and evaporated in vacuo. Column chromatography purification gave 5.33g (containing solvent) of oil in 110% yield with a small amount of solvent (SZY-1805-6H).
SZY-1805-6H (5.33 g,11.5mmol,1.0eq, MW463.52) was dissolved in 50ml DCM, to which DIPEA (4.46 g,34.5mmol,3eq, MW129.25) was added and the reaction stirred. Propionyl chloride (1.60 g,17.5mmol,1.5eq, MW92.52) was added in portions at 0℃over a period of about half an hour. After the completion of the low price, the reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 5.25g of oily liquid are obtained in 88% yield (SZY-1805-7H).
SZY-1805-7H (5 g) was dissolved in 20ml EA, and 50ml of 4M HCl.EA was added thereto, followed by stirring at room temperature. Every half hour of TLC monitoring, 1.5h-2h reaction was completed and no obvious impurity was shown. After the reaction, the solvent was removed by rotary evaporation to give an oily substance, which was neutralized with aqueous sodium hydrogencarbonate, dried over anhydrous sodium sulfate after extraction with ethyl acetate, and the solvent was distilled off to give 3g of a product (SZY 1805-1H).
The structural identification data are 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。
EXAMPLE 2,
Figure BDA0002496194080000081
Preparation of (SZY 1805-1 a) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 2:
to a 250ml round bottom flask was added SZY-1805-2a (10.0 g,31.22mmol,1.0eq, MW320.34), 100ml DMF was added to dissolve, imidazole (13.8 g,202.9mmol,6.5eq, MW68.08) was added to dissolve, then TBSCl (18.82 g,124.87mmol,4.0eq, MW150.7) was added in portions and the reaction was stirred at room temperature overnight. After completion of the TLC detection reaction, 150ml of EA and 200ml of water were added to the solution for delamination. The aqueous phase was extracted three times with 50ml ea and the organic phases were combined. The organic phase was washed with 1% hydrochloric acid solution and 3-5 times with pure water (removal of DMF). The organic phase was dried over anhydrous sodium sulfate and distilled off to give an oil. To the oil was added 50ml of THF, 50ml of water, 50ml of acetic acid, and the reaction was stirred at room temperature and monitored by TLC after 2h. To the reaction solution was added 150ml of EA and 200ml of water were separated, the aqueous phase was extracted 3 times with 50ml of EA, and the organic phases were combined. After washing the organic phase with water 5 times, it was dried over anhydrous sodium sulfate and distilled off to give a white solid (containing a small amount of solvent). PE (50 ml) was added to the mixture, followed by beating and suction filtration to obtain 12.0g of a white solid (SZY-1805-3 a) in a yield of 88.4%.
To a 250ml round bottom flask was added SZY-1805-3a (5.0 g,11.5mol,1.0eq, MW434.6) and dissolved by adding 50ml dichloromethane. SZY-1805-4a (1.5 g,13.8mmol,1.2eq,
MW 117.2), DMAP (150 mg,1.2mmol,0.1eq, MW122), EDCI (2.6 g,13.8mmol,1.2eq, MW191.7), and stirring at room temperature for 7h. After the completion of the TLC, DCM was removed by rotary evaporation under reduced pressure, and 150ml of EA was added and 150ml of water was separated. The organic phase was washed with 100ml of water 2 times, and with 100ml of saturated brine 2 times, dried over anhydrous sodium sulfate, and filtered and distilled to give 5.8g of an oil with a yield of 99% (SZY-1805-5 a).
To a 250ml round bottom flask containing SZY-1805-5a (5.8 g,11.5mmol,1.0eq, MW533.8) was added 50ml THF to dissolve, then TBAF (4.4 g,13.8mmol,1.2eq, MW315.5) was added and the reaction was stirred at room temperature. The solution became wine red after the addition of TBAF and then orange yellow. After 3h, TLC was monitored and the reaction was complete. To this was added 100ml of water and EA, and the layers were separated. The organic phase was washed with water, dried over anhydrous sodium sulfate and evaporated in vacuo. Column chromatography purification gave 4.0g (containing solvent) of oil in 108% yield (SZY-1805-6 a).
SZY-1805-6a (4.0 g,10.49mmol,1.0eq, MW419.5) was dissolved in 50ml DCM, to which DIPEA (4.07 g,31.47mmol,3eq, MW129.25) was added and the reaction stirred. Propionyl chloride (1.45 g,15.73mmol,1.5eq, MW92.52) was added in portions at 0℃over half an hour. After the completion of the low price, the reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 3.0g of oily liquid are obtained in 65% yield (SZY 1805-1 a).
The structural identification data are 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。
EXAMPLE 3,
Figure BDA0002496194080000091
Preparation of (SZY 1805-1 f) compounds the above compounds were prepared according to the synthetic scheme shown in fig. 3:
to a 250ml round bottom flask was added SZY-1805-2f (10.0 g,31.22mmol,1.0eq, MW320.34), 100ml DMF was added to dissolve, imidazole (13.8 g,202.9mmol,6.5eq, MW68.08) was added to dissolve, then TBSCl (18.82 g,124.87mmol,4.0eq, MW150.7) was added in portions and the reaction was stirred at room temperature overnight. After completion of the TLC detection reaction, 150ml of EA and 200ml of water were added to the solution for delamination. The aqueous phase was extracted three times with 50ml ea and the organic phases were combined. The organic phase was washed with 1% hydrochloric acid solution and 3-5 times with pure water (removal of DMF). The organic phase was dried over anhydrous sodium sulfate and distilled off to give an oil. To the oil was added 50ml of THF, 50ml of water, 50ml of acetic acid, and the reaction was stirred at room temperature and monitored by TLC after 2h. To the reaction solution was added 150ml of EA and 200ml of water were separated, the aqueous phase was extracted 3 times with 50ml of EA, and the organic phases were combined. After washing the organic phase with water 5 times, it was dried over anhydrous sodium sulfate and distilled off to give a white solid (containing a small amount of solvent). PE50ml was added to the mixture, and the mixture was beaten and filtered to obtain 12.0g of a white solid with a yield of 88.4% (SZY-1805-3 f).
To a 250ml round bottom flask was added SZY-1805-3f (5.0 g,11.5mol,1.0eq, MW434.6) and dissolved by adding 50ml dichloromethane. SZY1805-4f (1.6 g,13.8mmol,1.2eq, MW117.2), DMAP (150 mg,1.2mmol,0.1eq, MW122), EDCI (2.6 g,13.8mmol,1.2eq, MW191.7) were added sequentially and the reaction was stirred at room temperature for 7h. After the completion of the TLC, DCM was removed by rotary evaporation under reduced pressure, and 150ml of EA was added and 150ml of water was separated. The organic phase was washed with 100ml of water 2 times, with 100ml of saturated brine 2 times, dried over anhydrous sodium sulfate, and filtered and distilled to give 6.1g of an oil with a yield of 99% (SZY-1805-5 f).
To a 250ml round bottom flask containing SZY1805-5f (6.1 g,11.5mmol,1.0eq, MW533.8) was added 50ml THF to dissolve, then TBAF (4.4 g,13.8mmol,1.2eq, MW315.5) was added and the reaction was stirred at room temperature. The solution became wine red after the addition of TBAF and then orange yellow. After 3h, TLC was monitored and the reaction was complete. To this was added 100ml of water and EA, and the layers were separated. The organic phase was washed with water, dried over anhydrous sodium sulfate and evaporated in vacuo. Column chromatography purification gave 4.4g (containing solvent) of oil in 108% yield (SZY-1805-6 f).
SZY-1805-6f (4.4 g,10.49mmol,1.0eq, MW419.5) was dissolved in 50ml DCM, to which DIPEA (4.07 g,31.47mmol,3eq, MW129.25) was added and the reaction stirred. Propionyl chloride (1.45 g,15.73mmol,1.5eq, MW92.52) was added in portions at 0℃over half an hour. After the completion of the low price, the reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 3.22g of oily liquid are obtained in 65% yield (SZY 1805-1 f).
The structural identification data are 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。
EXAMPLE 4,
Figure BDA0002496194080000111
Preparation of (SZY 1805-1 b) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 4:
to a 250ml round bottom flask was added SZY-1805-2b (10.0 g,31.22mmol,1.0eq, MW320.34), 100ml DMF was added to dissolve, imidazole (13.8 g,202.9mmol,6.5eq, MW68.08) was added to dissolve, then TBSCl (18.82 g,124.87mmol,4.0eq, MW150.7) was added in portions and the reaction was stirred at room temperature overnight. After completion of the TLC detection reaction, 150ml of EA and 200ml of water were added to the solution for delamination. The aqueous phase was extracted three times with 50ml ea and the organic phases were combined. The organic phase was washed with 1% hydrochloric acid solution and 3-5 times with pure water (removal of DMF). The organic phase was dried over anhydrous sodium sulfate and distilled off to give an oil. To the oil was added 50ml of THF, 50ml of water, 50ml of acetic acid, and the reaction was stirred at room temperature and monitored by TLC after 2h. To the reaction solution was added 150ml of EA and 200ml of water were separated, the aqueous phase was extracted 3 times with 50ml of EA, and the organic phases were combined. After washing the organic phase with water 5 times, it was dried over anhydrous sodium sulfate and distilled off to give a white solid (containing a small amount of solvent). PE50ml was added to the mixture, and the mixture was beaten and filtered to obtain 12.0g of a white solid with a yield of 88.4% (SZY-1805-3 b).
To a 250ml round bottom flask was added SZY-1805-3b (5.0 g,11.5mol,1.0eq, MW434.6) and dissolved by adding 50ml dichloromethane. SZY-1805-4b (1.6 g,13.8mmol,1.2eq,
MW 117.2), DMAP (150 mg,1.2mmol,0.1eq, MW122), EDCI (2.6 g,13.8mmol,1.2eq, MW191.7), and stirring at room temperature for 7h. After the completion of the TLC, DCM was removed by rotary evaporation under reduced pressure, and 150ml of EA was added and 150ml of water was separated. The organic phase was washed with 100ml of water 2 times, with 100ml of saturated brine 2 times, dried over anhydrous sodium sulfate, and filtered and distilled to give 6.1g of an oil with a yield of 99% (SZY-1805-5 b).
To a 250ml round bottom flask containing SZY1805-5b (6.1 g,11.5mmol,1.0eq, MW533.8) was added 50ml THF to dissolve, then TBAF (4.4 g,13.8mmol,1.2eq, MW315.5) was added and the reaction was stirred at room temperature. The solution became wine red after the addition of TBAF and then orange yellow. After 3h, TLC was monitored and the reaction was complete. To this was added 100ml of water and EA, and the layers were separated. The organic phase was washed with water, dried over anhydrous sodium sulfate and evaporated in vacuo. Column chromatography purification gave 4.4g (containing solvent) of oil in 108% yield (SZY-1805-6 b).
SZY-1805-6b (4.4 g,10.49mmol,1.0eq, MW419.5) was dissolved in 50ml DCM, to which DIPEA (4.07 g,31.47mmol,3eq, MW129.25) was added and the reaction stirred. Dimethylaminoacyl chloride (1.6 g,15.73mmol,1.5eq, MW92.52) was added in portions at 0deg.C, over half an hour. After the completion of the low price, the reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 3.5g of oily liquid are obtained in 70% yield (SZY 1805-1 b).
The structural identification data are 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。
EXAMPLE 5,
Figure BDA0002496194080000121
Preparation of (SZY 1805-1 c) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 5:
to a 250ml round bottom flask was added SZY-1805-6 (10.0) and dissolved by adding 50ml dichloromethane. SZY-1805-7c (5 g), DMAP (0.2 g), EDCI (5 g) were added in this order, and the reaction was stirred at room temperature for 7 hours. After the completion of the TLC, DCM was removed by rotary evaporation under reduced pressure, and 150ml of EA was added and 150ml of water was separated. The organic phase was washed with 100ml of water 2 times, and with 100ml of saturated brine 2 times, dried over anhydrous sodium sulfate, and filtered and distilled to dryness to give 8.0g of an oil (SZY-1805-8 c).
SZY-1805-8c (8.0 g) was dissolved in 40ml of EA, 40ml of 4M HCl.EA was added thereto, and the reaction was stirred at room temperature. Every half hour of TLC monitoring, 1.5h-2h reaction was completed and no obvious impurity was shown. After the reaction, the solvent was removed by rotary evaporation to give an oily substance, which was neutralized with aqueous sodium hydrogencarbonate, dried over anhydrous sodium sulfate after extraction with ethyl acetate, and the solvent was distilled off to give 6.2g of a product (SZY 1805-1 c).
The structural identification data are 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。
EXAMPLE 6,
Figure BDA0002496194080000131
/>
Preparation of (SZY 1805-1 d) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 6:
SZY-1805-6 (10 g) was dissolved in 50ml of DCM, to which DIPEA (4.07 g,31.47mmol,3eq, MW129.25) was added and the reaction stirred. Tertiary pentanoyl chloride (2.0 g) was added in portions at 0℃over about half an hour. After the completion of the low price, the reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 8.0g of oily liquid are obtained in 80% yield (SZY 1805-1 d).
The structural identification data are 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,
Figure BDA0002496194080000132
Preparation of (SZY 1805-1 e) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 7:
to a 250ml round bottom flask was added SZY-1805-6 (10.0) and dissolved by adding 50ml dichloromethane. SZY-1805-7e (6 g), DMAP (0.2 g), EDCI (5 g) were added in this order, and the reaction was stirred at room temperature for 12 hours. After the completion of the TLC, DCM was removed by rotary evaporation under reduced pressure, and 150ml of EA was added and 150ml of water was separated. The organic phase was washed with 100ml of water 2 times, and with 100ml of saturated brine 2 times, dried over anhydrous sodium sulfate, and filtered and distilled to dryness to give 9.0g of an oil (SZY-1805-8 e).
SZY-1805-8e (9.0 g) was dissolved in 50ml of EA, and 50ml of 4M HCl.EA was added thereto, followed by stirring at room temperature. Every half hour of TLC monitoring, 1.5h-2h reaction was completed and no obvious impurity was shown. After the reaction, the solvent was removed by rotary evaporation to give an oily substance, which was neutralized with aqueous sodium hydrogencarbonate, dried over anhydrous sodium sulfate after extraction with ethyl acetate, and the solvent was distilled off to give 6.0g of a product (SZY 1805-1 e).
The structural identification data are 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,
Figure BDA0002496194080000141
Preparation of (SZY 1805-1 k) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 8:
SZY-1805-6 (10 g) was dissolved in 50ml of DCM, to which DIPEA (6 g,31.47mmol,3eq, MW129.25) was added and the reaction stirred. N-hexanoyl chloride (5.0 g) was added in portions at 0℃over about half an hour. After the completion of the low price, the reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 8.5g of oily liquid are obtained in 81% yield (SZY 1805-1 k).
The structural identification data are 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。
EXAMPLE 9,
Figure BDA0002496194080000151
Preparation of (SZY 1805-1M) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 9:
SZY-1805-6 (10 g) was dissolved in 50ml of DCM, to which DIPEA (6 g,31.47mmol,3eq, MW129.25) was added and the reaction stirred. Succinic anhydride (5.0 g) was added at 0deg.C. The reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 7.0g of oily liquid are obtained in 68% yield (SZY-1805-1M).
The structural identification data are 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。
EXAMPLE 10,
Figure BDA0002496194080000152
/>
Preparation of (SZY 1805-1L) Compounds
SZY-1805-6H (5.33 g,11.5mmol,1.0eq, MW463.52) was dissolved in 50ml DCM, to which DIPEA (4.46 g,34.5mmol,3eq, MW129.25) was added and the reaction stirred. Acetyl chloride (1.50 g,17.5mmol,1.5eq, MW92.52) was added in portions at 0℃over a half hour period. After the completion of the low price, the reaction was stirred at room temperature overnight. The next day TLC monitored the reaction was complete. The reaction mixture was washed with 100ml of water 2 times and 100ml of saturated brine 2 times. The organic phase was dried over anhydrous sodium sulfate, distilled off, and separated by column chromatography. 5.00g of oily liquid are obtained in 80% yield (SZY 1805-7L).
SZY-1805-7L (5 g) was dissolved in 20ml EA, and 50ml of 4M HCl.EA was added thereto, followed by stirring at room temperature. Every half hour of TLC monitoring, 1.5h-2h reaction was completed and no obvious impurity was shown. After the reaction, the solvent was removed by rotary evaporation to give an oily substance, which was neutralized with aqueous sodium hydrogencarbonate, dried over anhydrous sodium sulfate after extraction with ethyl acetate, and the solvent was distilled off to give 2.8g (SZY 1805-1L) of a product.
The structural identification data are 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
EXAMPLE 11,
Figure BDA0002496194080000161
Preparation of (SZY 1805-1 p) Compounds
Compound SZY1805-1p HCl is 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 (100.0 g,312.2mmol,1.0 eq), 600mL DMF was added to dissolve, imidazole (95.65 g,1.45mmol,4.5 eq) was added and the reaction was stirred for 0.5h, then TBSCl (117.61 g,780.4mmol,2.5 eq) was added in portions and stirred at room temperature overnight.
The next day TLC detected completion of reaction (n-heptane: ethyl acetate=1:2, rf 1 ＝0.8，Rf 2 =0.3), adding 3.0L of water to the solution, adding EA 300ml of x 4 extraction, combining concentrated organic phases; 3.0L of water was added to the organic phase, extracted with 300ml of EA, and the concentrated organic phases were combined; 3.0L of water was added to the organic phase, extracted with 300ml of EA, and the combined spin-on was used to remove the majority of the organic phase and then carried on directly to the next step.
2: SZY1805-4p synthesis
400ml of THF,400ml of water and 400ml of acetic acid were added to the product of the previous step, and the reaction was stirred for about 2 hours, and the reaction was completed by TLC (n-heptane: ethyl acetate=1:2, rf=0.3). Adding 3L of water into the reaction solution, adding 400ml of DCM (data center) for extraction, and combining concentrated organic phases; 3L of water was added to the organic phase, 400ml of DCM 3 was added for extraction, and the concentrated organic phases were combined; 3L of water was added to the organic phase, 400ml of DCM 3 was added for extraction, and the organic phases were combined and dried to give a white solid powder. The white solid powder was slurry-washed with 500ml of n-heptane, suction-filtered and dried to obtain 129g of white solid powder in 95% yield.
3: SZY1805-5p synthesis
SZY1805-4p (36.82 g,84.72mmol,1.0 eq) was dissolved in 360ml DCM and Boc aminopropanol, EDCI, DMAP were added sequentially to the reaction flask and the reaction was stirred. After 5h of reaction, TLC detection was complete (n-heptane: ethyl acetate=1:2, rf=0.8). The reaction is directly carried out for the next reaction after removing part of the organic solvent by rotary evaporation.
4: SZY1805-6p synthesis
500ml of THF was added to the previous reaction, and TBAF.3H was added 2 O (29.4 g,93.19mmol,1.1 eq) was stirred for 1h and the reaction was completed by TLC. 400ml of 1% aqueous HCl was added to the reaction, extracted with 100ml of x 4 DCM and the organic phases combined; the organic phase was washed with 400ml of 1% aqueous hcl, the aqueous phase was extracted with 100ml x 4 DCM and the concentrated organic phases were combined; the organic phase was washed with 400ml of 1% aqueous hcl, the aqueous phase was extracted with 100ml x 4 DCM and the dried organic phases were combined to give about 40.46g of a colorless oil in 100% yield.
5: SZY1805-7p Synthesis
SZY1805-6p (40.46 g,84.72mmol,1.0 eq) was dissolved in 400ml DCM and DIPEA (32.85 g,254.16mmol,3.0 eq) was added under ice-water bath and stirred for 0.5h. Propionyl chloride (11.76 g,127.08mmol,1.5 eq) was then added in portions and the reaction stirred overnight. After the completion of the reaction by TLC the next day 400ml of water was added to the reaction and extracted with DCM100ml x 4 times and the organic phases combined. The organic phase was washed with 400ml water and the aqueous phase extracted with 100ml x 4 times DCM and the concentrated organic phases combined; the organic phase was washed successively with 400ml of water, the aqueous phase extracted 100ml x 4 times with DCM and the spin-dried phases were combined to give about 45g of a reddish brown oil in 96% yield. (HPLC monitoring, column chromatography purification if the purity is low)
6: SZY1805-1p HCl synthesis
SZY1805-7p (45 g) was dissolved in 400ml of 2mol/L EA.HCl and reacted for 1 hour under stirring at normal temperature, a large amount of white solid powder was precipitated, and the filter cake was collected by filtration. The filter cake was washed with 200ml EA slurry for 1h and collected by suction filtration. Repeated washing with EA slurry for three times, filtering and baking the material to obtain 25g of white solid powder, with the yield of 71.4% and the HPLC purity of 98.9%.
The structural identification data are 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.
EXAMPLE 12,
Figure BDA0002496194080000171
Preparation of (SZY 1805-1 q) Compounds
The above compounds were prepared according to the synthetic scheme shown in fig. 11:
the specific operation is as follows:
1: SZY1805-3q Synthesis
To a 1000mL round bottom flask was added SZY1805-2q (100.0 g,312.2mmol,1.0 eq), 600mL DMF was added to dissolve, imidazole (95.65 g,1.45mmol,4.5 eq) was added and the reaction was stirred for 0.5h, then TBSCl (117.61 g,780.4mmol,2.5 eq) was added in portions and stirred at room temperature overnight.
The next day TLC detected completion of reaction (n-heptane: ethyl acetate=1:2, rf 1 ＝0.8，Rf 2 =0.3), adding 3.0L of water to the solution, adding EA 300ml of x 4 extraction, combining concentrated organic phases; 3.0L of water was added to the organic phase, extracted with 300ml of EA, and the concentrated organic phases were combined; 3.0L of water was added to the organic phase, extracted with 300ml of EA, and the combined spin-on was used to remove the majority of the organic phase and then carried on directly to the next step.
2: SZY1805-4q synthesis
400ml of THF,400ml of water and 400ml of acetic acid were added to the product of the previous step, and the reaction was stirred for about 2 hours, and the reaction was completed by TLC (n-heptane: ethyl acetate=1:2, rf=0.3). Adding 3L of water into the reaction solution, adding 400ml of DCM (data center) for extraction, and combining concentrated organic phases; 3L of water was added to the organic phase, 400ml of DCM 3 was added for extraction, and the concentrated organic phases were combined; 3L of water was added to the organic phase, 400ml of DCM 3 was added for extraction, and the organic phases were combined and dried to give a white solid powder. The white solid powder was slurry-washed with 500ml of n-heptane, suction-filtered and dried to obtain 125g of white solid powder in 97% yield.
3: SZY1805-5q Synthesis
SZY1805-4q (36.82 g,84.72mmol,1.0 eq) was dissolved in 360ml DCM and Boc aminoethylamine, EDCI, DMAP were added sequentially to the reaction flask and the reaction was stirred. After 5h of reaction, TLC detection was complete (n-heptane: ethyl acetate=1:2, rf=0.8). The reaction is directly carried out for the next reaction after removing part of the organic solvent by rotary evaporation.
4: SZY1805-6q Synthesis
500ml of THF was added to the previous reaction, and TBAF.3H was added 2 O (29.4 g,93.19mmol,1.1 eq) was stirred for 1h and the reaction was completed by TLC. 400ml of 1% aqueous HCl was added to the reaction, extracted with 100ml of x 4 DCM and the organic phases combined; the organic phase was washed with 400ml of 1% aqueous hcl, the aqueous phase was extracted with 100ml x 4 DCM and the concentrated organic phases were combined; the organic phase was washed with 400ml of 1% aqueous hcl, the aqueous phase was extracted with 100ml x 4 DCM and the dried organic phases were combined to give about 39.02g of a colorless oil in 98% yield.
5: SZY1805-7q Synthesis
SZY1805-6q (39.02 g,84.72mmol,1.0 eq) was dissolved in 400ml DCM and DIPEA (32.85 g,254.16mmol,3.0 eq) was added under ice-water bath and stirred for 0.5h. Propionyl chloride (11.76 g,127.08mmol,1.5 eq) was then added in portions and the reaction stirred overnight. After the completion of the reaction by TLC the next day 400ml of water was added to the reaction and extracted with DCM100ml x 4 times and the organic phases combined. The organic phase was washed with 400ml water and the aqueous phase extracted with 100ml x 4 times DCM and the concentrated organic phases combined; the organic phase was washed successively with 400ml of water, the aqueous phase extracted 100ml x 4 times with DCM and the spin-dried phases were combined to give about 42g of a reddish brown oil in 92% yield. (HPLC monitoring, column chromatography purification if the purity is low)
6: SZY1805-1q HCl synthesis
SZY1805-7q (42 g) was dissolved in 400ml 2mol/L EA.HCl and reacted for 1 hour under stirring at normal temperature, a large amount of white solid powder was precipitated, and the filter cake was collected by filtration. The filter cake was washed with 200ml EA slurry for 1h and collected by suction filtration. Repeated washing with EA slurry for three times, filtering and baking the material to obtain 28g of white solid powder, with the yield of 73% and the HPLC purity of 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 can be prepared from each compound in ethyl acetate hydrochloride, and the related tartrate can be prepared by adding tartaric acid into acetone solution.
EXAMPLE 13 stability investigation
Tartrate salt of the compound prepared in example 3, the structural formula is as follows
Figure BDA0002496194080000191
Stability experiment: sZY1805-1f-JSS stability test statistical table under different solutions
Solution 0h(％) 24h(％) 72h(％) 5d(％) Remarks
Deionized water 99.25 99.15 98.78 98.72
Physiological saline 99.25 99.17 98.85 98.49
phosphate buffer at pH6.3 97.76 92.17 76.62
phosphate buffer at pH6.5 98.85 96.37 90.33 With overlap of
phosphate buffer at pH6.8 93.98 77.17 42.90
phosphate buffer pH7.0 90.97 68.93 28.96
Note that: purity was liquid phase relative purity (HPLC) at 254nm. The pH6.5 product peak overlaps with the impurity peak, and the undivided product peak causes a tendency to be false.
The hydrochloride salt of the compound prepared in example 1 has the following structural formula:
Figure BDA0002496194080000201
SZY1805-1H stability test statistics table for different solutions:
solution 0h(％) 24h(％) 72h(％) 5d(％) Remarks
Deionized water 98.99 98.70 97.67 97.06
Physiological saline 99.01 98.92 98.70 98.05
phosphate buffer at pH6.3 97.77 86.38 60.80
phosphate buffer at pH6.5 93.89 74.21 44.13
phosphate buffer at pH6.8 92.17 59.71 19.52
phosphate buffer pH7.0 93.79 49.60 8.81
Note that: purity was liquid phase relative purity (HPLC) at 254nm.
Liquid phase conditions:
phase A: 0.05% phosphoric acid aqueous solution
And B phase: 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 injection amount: 10 mu L
Sample concentration: 1mg/mL
Detection wavelength: 254nm
Chromatographic column: agilent Zorbax SB C18 and 5218
The stability of the compounds prepared in the examples in aqueous physiological saline is shown in the following table (ph=6, storage temperature=25 ℃): (the following compounds are hydrochloride salts of the corresponding compounds except for sodium mycophenolate)
Chemical combinationArticle (B) 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
Mycophenolic acid sodium salt 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 tables show the results of stability experiments of mycophenolate sodium, SZY1805-1f and SZY1805-1L in water:
stability test of Meicosanate sodium (about 0.25mg in 1 ml)
Figure BDA0002496194080000211
SZY1805-1f (hydrochloride) stability experimental data: (about 0.25mg in 1 ml) stability test (Water)
Figure BDA0002496194080000212
SZY1805-1L stability experimental data: (about 0.25mg in 1 ml) stability test (Water)
Figure BDA0002496194080000221
The compound in the general formula has different physicochemical properties according to different structures, and the stability in water, physiological saline and buffer salt is different, so that the compound in the general formula is stable in general.
Example 14 pharmacokinetic experiments
Test sample mycophenolate sodium (mycophenolate sodium): white powdery solid, molecular weight: 342.32, lot number: BSP-20171123, purity: 99.0% and preservation conditions: sealing and storing at normal temperature, and the effective period is 2019.11.22.
Sample SZY1805-1f JSS: pale yellow viscous liquid, molecular weight: 625.66, purity: > 98%.
Sample SZY1805-1H HCl: white solid, molecular weight: 455.93, purity: 98.8%.
Detection of compounds: mycophenolic acid sodium salt
The eye drops were administered ocularly, and after each administration they helped gently occlude the double eyelid for about 10 seconds.
Dosing vehicle: sodium chloride injection
Equimolar dosing: 0.16. Mu.M/New Zealand rabbit/time; namely, the dosage of mycophenolate sodium group administered with mycophenolate sodium is 0.055 mg/New Zealand rabbit, the dosage of SZY1805-1f group administered with mycophenolate sodium is 0.1 mg/New Zealand rabbit, and the dosage of SZY1805-1H group administered with mycophenolate sodium is 0.073 mg/New Zealand rabbit
100 uL/New Zealand rabbit
SZY1805-1f JSS is 1mg/mL, SZY1805-1H HCl 0.73mg/mL, mycophenolate sodium is 0.55mg/mL
Lower limit of detection quantification: 10ng/mL
Results: 1) After equimolar (0.16 mu M/dose of New Zealand rabbits) ocular administration of SZY1805-1f JSS and SZY1805-1H HCl, respectively, active metabolite mycophenolate sodium can be detected in aqueous humor of the New Zealand rabbits; 2) In rabbit aqueous humor after 1H administration, the contents of the sodium mycophenolate in the SZY1805-1f group and the sodium mycophenolate in the SZY1805-1H group are close to each other and are 724.4 +/-313.6 ng/ml and 730.6 +/-434.0 ng/ml respectively, and are higher than 180.0+/-72.2 ng/ml of the sodium mycophenolate group; 3) At this concentration of SZY1805-1f JSS, SZY1805-1H HCl, no irritation was produced to eyes of New Zealand rabbits.
After eye administration of 0.16 mu M/SZY 1805-1f JSS, SZY1805-1H HCl and mycophenolate sodium in New Zealand rabbits for 1H, the concentration of mycophenolate sodium in aqueous humor (ng/ml)
Figure BDA0002496194080000222
Figure BDA0002496194080000231
The following compound local pharmacokinetic experiments were performed according to the above method, and the experimental results are as follows:
after eye administration of New Zealand rabbits at 0.16. Mu.M/dose, the concentration of mycophenolate sodium in aqueous humor (ng/ml)
Figure BDA0002496194080000232
After eye administration of New Zealand rabbits at 0.16. Mu.M/dose, the concentration of mycophenolate sodium in aqueous humor (ng/ml)
Figure BDA0002496194080000233
Note that: the above compounds are all corresponding hydrochloride
Figure BDA0002496194080000234
Figure BDA0002496194080000241
Example 15, example Compound eye drops Rabbit eye irritation test
The purpose is as follows: eye drops prepared from the above example compounds at different concentrations were observed for irritation to rabbit eyes.
The method comprises the following steps: 165 male New Zealand rabbits, with no secretion in both eyes, no redness in conjunctiva, and no abnormality in cornea and iris, were divided into 55 groups, and 3 animals/group, and 0.025%, 0.05%, 0.1%, 0.2%, and 0.4% of the compound of the examples (SZY 1805-1a, SZY1805-1b, SZY1805-1c, SZY1805-1d, SZY1805-1e, SZY1805-1k, SZY1805-1M, SZY1805-1f, SZY1805-1H, SZY1805-1p, and SZY1805-1 q) were respectively administered as eye drops. Using the autologous left and right side comparison method, the eyes of each group of animals were given eye drops of the compound of the example at the corresponding concentrations, and the eyes were given an equivalent amount of physiological saline, 50. Mu.l/eye. Dosing d1 was 1 time, followed by 3 times daily dosing (about 4 hours apart) for 14 consecutive days (d 1-d 14). 1, 2, 4, 24h after the first administration, 1, 2h before each administration, 1, 2, 4, 24, 48, and 72h after the last administration were observed for rabbit eye irritation and scores were recorded, and finally scoring was performed according to the eye irritation evaluation criteria. At the end of the test, 3 animal eyeballs of the compound eye drop group of example at a concentration of 0.4% and other animal eyeballs having abnormalities were taken for histopathological examination.
Results: the administration is continued for 14 days, and visual observation results show that the secretion of rabbit eyes is increased in each group of administration sides compared with the control sides during the administration period, conjunctiva is free from congestion and edema, cornea is transparent and free from turbidity, iris texture is clear, congestion and swelling are avoided, and eye irritation scores of all animals are 0-2 minutes. Each group of observations was rated less than 3 points for no irritation (eye irritation rating 0-3 points: no irritation).
Conclusion: the eye drops of the compound of the examples with the concentration of 0.025%, 0.05%, 0.1%, 0.2% and 0.4% have no obvious irritation to normal rabbit eyes after continuous administration for 14 days.
Note that: animal experiments were not completed, administration was completed for 14 days, and the period was observed. In view of the detailed observation, 3 animal eyeballs of the compound eye drop group of example with a concentration of 0.4% and other animal eyeballs with abnormalities were extracted after the end of the observation of 72 hours after the last administration, and pathology was sent to conduct histopathological examination.

Claims (2)

1. A compound selected from:
Figure QLYQS_1
/>
Figure QLYQS_2
2. an ophthalmic liquid formulation comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof, a solvent, one or more physiologically acceptable excipients;
the solvent is water;
the ocular liquid formulation is free of added co-solvents.
CN202010419146.5A 2019-08-20 2020-05-18 Prodrugs of mycophenolic acid and method for preparing same Active CN111393399B (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN2019107672977 2019-08-20
CN201910767297.7A CN110526885A (en) 2019-08-20 2019-08-20 Pro-drug of Mycophenolic Acid and preparation method thereof

Publications (2)

Publication Number Publication Date
CN111393399A CN111393399A (en) 2020-07-10
CN111393399B true CN111393399B (en) 2023-05-30

Family

ID=68662471

Family Applications (2)

Application Number Title Priority Date Filing Date
CN201910767297.7A Pending CN110526885A (en) 2019-08-20 2019-08-20 Pro-drug of Mycophenolic Acid and preparation method thereof
CN202010419146.5A Active CN111393399B (en) 2019-08-20 2020-05-18 Prodrugs of mycophenolic acid and method for preparing same

Family Applications Before (1)

Application Number Title Priority Date Filing Date
CN201910767297.7A Pending CN110526885A (en) 2019-08-20 2019-08-20 Pro-drug of Mycophenolic Acid and preparation method thereof

Country Status (2)

Country Link
CN (2) CN110526885A (en)
WO (1) WO2021031999A1 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110526885A (en) * 2019-08-20 2019-12-03 上海英诺富成生物科技有限公司 Pro-drug of Mycophenolic Acid and preparation method thereof
CN115057837A (en) * 2022-07-01 2022-09-16 上海英诺富成生物科技有限公司 Benzofuran compound and preparation method and application thereof

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4727069A (en) * 1987-01-30 1988-02-23 Syntex (U.S.A.) Inc. Heterocyclic aminoalkyl esters of mycophenolic acid, derivatives thereof and pharmaceutical compositions

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4725622A (en) * 1986-01-23 1988-02-16 Syntex (U.S.A.) Inc. Mycophenolic acid derivatives in the treatment of rheumatoid arthritis
US4753935A (en) * 1987-01-30 1988-06-28 Syntex (U.S.A.) Inc. Morpholinoethylesters of mycophenolic acid and pharmaceutical compositions
JPH0930966A (en) * 1995-07-24 1997-02-04 Gakuzo Tamura New pharmaceutical preparation for eye
CN102952105A (en) * 2011-08-26 2013-03-06 中国科学院上海药物研究所 Mycophenolic acid derivative, preparation method thereof and application thereof
CN110526885A (en) * 2019-08-20 2019-12-03 上海英诺富成生物科技有限公司 Pro-drug of Mycophenolic Acid and preparation method thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4727069A (en) * 1987-01-30 1988-02-23 Syntex (U.S.A.) Inc. Heterocyclic aminoalkyl esters of mycophenolic acid, derivatives thereof and pharmaceutical compositions

Also Published As

Publication number Publication date
WO2021031999A1 (en) 2021-02-25
CN110526885A (en) 2019-12-03
CN111393399A (en) 2020-07-10

Similar Documents

Publication Publication Date Title
EP0846685B1 (en) 3-Hydroxypyridine-2-carboxylic acid amide esters, their preparation and their use as medicaments
AU2016349080A1 (en) 7-(thiazol-5-yl) pyrrolopyrimidine compound as TLR7 agonist
CN111393399B (en) Prodrugs of mycophenolic acid and method for preparing same
US20150166530A1 (en) Crystalline Salts of a Potent HCV Inhibitor
FR2518088A1 (en) NOVEL AMINOACID DERIVATIVES AND THEIR THERAPEUTIC APPLICATION
WO2013186384A1 (en) N-substituted second generation derivatives of antifungal antibiotic amphotericin b and methods of their preparation and application
RU2736216C1 (en) Pharmaceutically acceptable salt of alkylcarbamoylnaphthalenyloxyoxyctenoyl hydroxy amide or derivative thereof and method for production thereof
KR20160067840A (en) Peptide-oligourea chimeric compounds and methods of their use
CN112047993A (en) Alpha-glucosidase inhibitor and application thereof
EP1453803A2 (en) Tetrahydrocarbozole derivatives as ligands for g-protein coupled receptors (gpcr)
DE69923460T2 (en) METHOD FOR PRODUCING RING-MODIFIED CYCLIC PEPTIDE ANALOGS
EP0314275A2 (en) 2-Pyrrolidone-5-carboxylic acid compounds
CN104974221B (en) Dipeptide and tripeptide proteasome inhibitors and preparation method and pharmaceutical application thereof
DE60206376T2 (en) FACTOR XA INHIBITOR
EP2917204B1 (en) Derivatives of 1h-indole-3-carboxamide and their use as p2y12 antagonists
CN101671367A (en) Pleuromutilin phosphate compounds, medicinal composition thereof, preparation method thereof and application thereof
CN112218878A (en) NTCP inhibitors
CN114685519B (en) Pyranocarbazole alkaloid derivatives and application thereof in treating Alzheimer's disease
DE1958383C3 (en) Leupeptins, processes for their preparation and pharmaceuticals containing them
CN115960080B (en) Polycyclic compounds and their use as ghrelin receptor agonists
JP5271904B2 (en) Aminoisoquinoline thrombin inhibitors with improved bioavailability
CN113234013B (en) Compound for inhibiting collagen synthesis and deposition and application thereof
EP0684257A1 (en) Pseudopeptide derivatives of neurokinines, process for their preparation and pharmaceutical compositions comprising them
CN107903245B (en) A kind of compound and its application in preparing treatment medicine for treating rheumatoid arthritis
JP4493891B2 (en) New derivatives of echinocandin, their production process and their use as antibacterial agents

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
TA01 Transfer of patent application right

Effective date of registration: 20230307

Address after: 511400 Room 402, No. 223, Huanshi Avenue West, Nansha District, Guangzhou, Guangdong

Applicant after: Guangzhou runer eye Biotechnology Co.,Ltd.

Address before: 201516 building 7, 228 jingle Road, Langxia Town, Jinshan District, Shanghai (Langxia economic community)

Applicant before: Shanghai innofucheng Biotechnology Co.,Ltd.

TA01 Transfer of patent application right
GR01 Patent grant
GR01 Patent grant