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CN118047811A - Anti-influenza virus phosphate compound and application thereof - Google Patents

Anti-influenza virus phosphate compound and application thereof Download PDF

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Publication number
CN118047811A
CN118047811A CN202211395789.6A CN202211395789A CN118047811A CN 118047811 A CN118047811 A CN 118047811A CN 202211395789 A CN202211395789 A CN 202211395789A CN 118047811 A CN118047811 A CN 118047811A
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compound
formula
atom
pharmaceutically acceptable
salt
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请求不公布姓名
侯雯
石江涛
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Shijiazhuang Dikaiwei Pharmaceutical Technology Co ltd
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Shijiazhuang Dikaiwei Pharmaceutical Technology Co ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
    • C07F9/65616—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings containing the ring system having three or more than three double bonds between ring members or between ring members and non-ring members, e.g. purine or analogs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6564—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
    • C07F9/6571—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms
    • C07F9/6574—Esters of oxyacids of phosphorus
    • C07F9/65742—Esters of oxyacids of phosphorus non-condensed with carbocyclic rings or heterocyclic rings or ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6564—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
    • C07F9/6571—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus and oxygen atoms as the only ring hetero atoms
    • C07F9/6574—Esters of oxyacids of phosphorus
    • C07F9/65744—Esters of oxyacids of phosphorus condensed with carbocyclic or heterocyclic rings or ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs

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  • Pulmonology (AREA)
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Abstract

The invention discloses an anti-influenza virus phosphate compound and application thereof; the phosphate compound is a compound shown in the following formula (I) or a hydrate, solvate, optical isomer, polymorph, isotope derivative and pharmaceutically acceptable salt thereof. The compound of the invention can be used for preparing anti-influenza virus medicines.

Description

Anti-influenza virus phosphate compound and application thereof
Technical Field
The invention relates to phosphate compounds with anti-influenza virus activity or hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof, a preparation method thereof and application thereof in anti-influenza virus.
Background
Influenza viruses mainly include four types of influenza a virus, influenza b virus, influenza c virus and influenza d virus. Among them, influenza a virus and influenza b virus are major human influenza viruses, influenza a virus is the strongest among them, the number of people who are infected in the seasons of influenza is the largest, and serious respiratory tract infection can be induced, resulting in death of more than 30 tens of thousands of people worldwide from influenza each year.
Currently, the major anti-influenza drugs on the market are: amantadine (AMANTADINE), the neuraminidase inhibitor oseltamivir (Oseltamivir) or Zanamivir (Zanamivir). RNA polymerase of influenza virus contains Cap-dependent endonuclease (Cap-DEPENDENT ENDONUCLEASE), and inhibition of the activity of Cap-dependent endonuclease can inhibit proliferation of virus, and various heterocyclic compounds have been used as Cap-dependent endonuclease inhibitors.
To date, baluo Sha Weizhi is the first drug to be marketed at this target, and there are also some other molecules that enter the clinical research stage. But including baluo Sha Weizhi, these molecules all exhibit low solubility and poor oral bioavailability, which makes them undesirable as influenza therapeutics. Particularly for severe influenza patients, intravenous administration may be required for rapid onset of action, but lower solubility limits their intravenous administration.
Thus, development of orally-injectable medicines for treating influenza is still urgent.
Disclosure of Invention
Unless specifically stated otherwise herein, the terms used herein are all the basic meanings commonly understood by those skilled in the art.
The invention provides a phosphate compound with an anti-influenza virus effect, which can realize oral administration and intravenous injection administration.
The compound has better solubility and stability, can realize intravenous injection administration, and has better advantage in the aspect of treatment of severe patients.
Unexpectedly, the compounds of the present invention have higher solubility and, at the same time, still have better oral bioavailability, making it possible to meet both injection and oral administration; in addition, the improvement of bioavailability is expected to reduce the dosage of oral administration, and further improve the safety of oral administration of the compound.
The invention provides a phosphate compound shown in the following formula (I) or hydrate, solvate, optical isomer, polymorph, isotope derivative and pharmaceutically acceptable salt thereof:
in formula (I), R a is selected from hydrogen or deuterium or methyl;
R b and R c are each independently selected from hydrogen, deuterium, or methyl, or R b and R c together with the attached carbon form cyclopropyl;
X 1 is an O atom or an S atom;
x 2 is a Se atom or an S atom;
n1 is 0, 1 or 2;
each R 1 or R 2 is independently selected from hydrogen or methyl;
R 3 and R 4 are each independently selected from the following groups: hydroxy, C1-C8 alkoxy, C3-C8 cycloalkoxy, C3-C8 heterocycloalkoxy, C6-C10 aryloxy, C7-C12 aralkyloxy, and R 3 and R 4 must not be both C1-C8 alkoxy when X 2 is S atom; or R 3 and R 4 together with the phosphorus atom to which they are attached, e.g A 5-7 membered ring of (2); wherein R 5、R6,R7、R8、R9、R10、R11、R12 and R 13 are each independently hydrogen or C1-C3 alkyl, or R 5 and R 6,R7 and R 8,R10 and R 11,R11 and R 12 each together with the carbon atom to which they are attached form an aromatic ring.
In some embodiments, the phosphate compounds provided by the invention or hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof are shown in formula (ii):
The definition of the substituent in the formula (II) is defined as the formula (I).
In some embodiments, the phosphate compounds provided by the invention or hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof are shown in formula (iii):
The definition of the substituent in the formula (III) is defined as in the formula (I).
In some embodiments, the phosphate compounds provided by the invention or hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof are shown in formula (iv):
The definition of the substituent in the formula (IV) is defined as the formula (I).
In embodiments of the present application, the solvate refers to a complex formed by the interaction of a compound with a pharmaceutically acceptable solvent, including aqueous methanol, ethanol, isopropanol, n-butanol, acetic acid, ethanolamine, ethyl acetate.
In embodiments of the present application, the C1-C3 alkyl refers to a saturated aliphatic hydrocarbon group containing 1 to 3 carbon atoms in the molecule, including but not limited to methyl, ethyl, propyl, isopropyl, cyclopropyl.
In embodiments of the present application, the C1-C8 alkoxy group refers to a group in which a saturated aliphatic hydrocarbon group having 1 to 8 carbon atoms in the molecule is interrupted by an oxygen atom at any reasonable position, and includes, but is not limited to, methoxy, ethoxy, propoxy, isopropoxy, isobutoxy, n-butoxy, 2-ethylethoxy, and the like.
In embodiments of the present application, the C3-C8 cycloalkoxy group refers to a monocyclic or fused polycyclic saturated or unsaturated cyclic hydrocarbyloxy group having 3 to 8 carbon atoms, including, but not limited to, cyclopropyloxy, cyclopentyloxy, bicyclo [3.1.0] hexyloxy, bicyclo [3.2.0] heptyloxy, and the like.
In an embodiment of the present application, the C3-C8 heterocycloalkyl group refers to a group in which a C3-C8 heterocycloalkyl group is attached to oxygen, and the C3-C8 heterocycloalkyl group refers to a saturated or unsaturated cyclic group having 3 to 8 carbon atoms and 1 to 4 hetero atoms in the molecule; the C3-C8 heterocycloalkyl group includes, but is not limited to, aziridinyl, tetrahydrothienyl, tetrahydropyrrolyl, piperidinyl, hexahydropyridazinyl, dihydropyridinyl, cyclopentylsulfanyl, morpholinyl, and the like.
In an embodiment of the present application, the aryloxy group of C6-C10 refers to a group having an aromatic ring composed of 6 to 10 carbon atoms bonded to an oxygen atom, including but not limited to phenoxy, naphthoxy.
In an embodiment of the present application, the C7-C12 aralkyloxy group refers to a group containing an aralkyl group of 7 to 12 carbon atoms attached to an oxygen atom, and includes, but is not limited to, benzyloxy, phenethyloxy and the like.
In some embodiments, R a is hydrogen; in some embodiments, R a is deuterium; in some embodiments, R a is methyl.
In some embodiments, R b and R c are both hydrogen; in some embodiments, R b and R c are both deuterium; in some embodiments, R b is hydrogen and R c is deuterium; in some embodiments, R b and R c are both methyl; in some embodiments, R b and R c together with the attached carbon form cyclopropyl; in some embodiments, R b is hydrogen and R c is methyl.
In some embodiments, n1 is 0; in some embodiments, n1 is 1; in some embodiments, n1 is 2.
In some embodiments, X 1 is an O atom; in some embodiments, X 1 is an S atom.
In some embodiments, X 1 is a Se atom; in some embodiments, X 1 is an S atom.
In an embodiment of the invention, each R 1 or R 2 is independently hydrogen or methyl.
In an embodiment of the invention, when X 2 is an S atom, R 3 and R 4 must not be both C1-C8 alkoxy groups;
In some embodiments, R 3 and R 4 are selected from the following groups: C1-C8 alkoxy, C3-C8 cycloalkoxy, C3-C8 heterocycloalkoxy; preferably, R 3 and R 4 are C1-C3 alkoxy groups;
In some embodiments, R 3 and R 4 are selected from the following groups: C6-C10 aryloxy, C7-C12 aralkyloxy; preferably, R 3 and R 4 are benzyloxy;
In some embodiments, R 3 is selected from the following groups: C1-C8 alkoxy, C3-C8 cycloalkoxy, C3-C8 heterocycloalkoxy, C6-C10 aryloxy, C7-C12 aralkyloxy, R 4 is hydroxy;
In some embodiments, R 3 and R 4 together with the phosphorus atom to which they are attached form, e.g 5-7 Membered ring of (2).
In some embodiments, R 5、R6,R7、R8、R9、R10、R11、R12 and R 13 are each independently hydrogen; in some embodiments, R 5、R6,R7、R8、R9、R10、R11、R12 and R 13 are each independently C1-C3 alkyl;
In some embodiments, R 5 and R 6,R7 and R 8,R10 and R 11,R11 and R 12 each together with the carbon atom to which they are attached form an aromatic ring, preferably a benzene ring;
in some specific embodiments, R 5 and R 6 together with the carbon atoms to which they are attached form a benzene ring;
In some specific embodiments, R 7 and R 8 together with the carbon atoms to which they are attached form a benzene ring;
In some specific embodiments, R 10 and R 11 together with the carbon atoms to which they are attached form a benzene ring;
In some specific embodiments, R 11 and R 12 together with the attached carbon atom form a benzene ring.
In some specific embodiments, in formulas (I) through (iii), where R 4 is hydroxy, the compound may or may not form a salt; in some specific embodiments, in formulas (I) through (iii), where R 4 is hydroxy, the compound may form a salt with an alkali metal, alkaline earth metal, zinc ion, organic amine, basic amino acid, preferably where R 4 is hydroxy, the compound may form a sodium salt, potassium salt, magnesium salt, zinc salt, amine salt, basic amino acid salt.
The salt-forming compound has better crystallinity and solubility.
In an embodiment of the present invention, formula (I) contains a chiral center (represented by carbon atom), and the optical isomers refer to optical isomers caused by different configurations of the represented carbon atoms, and the compound of the present invention or the intermediate thereof may be obtained into a single configuration compound by chiral separation.
In some specific embodiments, the compounds of the invention are racemates; in some specific embodiments, the compounds of the present invention are in the S-configuration.
In an embodiment of the invention, after absolute configuration determination of the S-configuration compound by electron circular dichroism, optical rotation direction is determined by optical rotation test.
In an embodiment of the invention, compounds of the S-configuration and R-configuration were subjected to optical rotation test (according to the optical rotation assay of China pharmacopoeia 2020 edition-four-0621, with methanol as solvent), and compounds of the S-configuration were the levorotatory form.
The compounds provided by the present invention include, but are not limited to, the following:
Or a hydrate, solvate, optical isomer, polymorph, isotopic derivative, pharmaceutically acceptable salt thereof.
In another aspect of the present invention, there is provided a pharmaceutical composition comprising the above compound or a hydrate, solvate, optical isomer, polymorph, isotopic derivative, pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers include one or a combination of several fillers, binders, diluents, lubricants, preservatives, taste masking agents or co-solvents. The pharmaceutical composition can be used for resisting influenza virus.
Further, the dosage forms of the pharmaceutical composition are tablets, capsules, powder, granules, pills, suspensions, syrups and injection.
In a third aspect of the invention, the present invention provides the use of the above compounds, including hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, for combating influenza virus.
The invention provides application of the compound, including hydrate, solvate, optical isomer, polymorph, isotope derivative, pharmaceutically acceptable salt or pharmaceutical composition thereof in preparing anti-influenza virus drugs.
The present invention provides a method for preventing or treating influenza virus infection comprising administering to a subject in need thereof a therapeutically effective amount of a compound as described above, including hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof.
The compound has better solubility and stability and possibility of injection administration, and has better advantage in the aspect of treatment of severe patients.
Surprisingly, it was found in the study that the compounds of the present invention have a better oral bioavailability. It is believed that phosphoric acid compounds are highly polar and have low stability, while the phosphate groups are more liposoluble after ester formation, resulting in lower solubility, which leads to lower oral bioavailability. But the compound of the invention has higher solubility and simultaneously reduces the polarity of the compound, so that the compound can simultaneously meet the requirements of injection and oral administration, and has extremely high development value.
In addition, the improvement of bioavailability is expected to reduce the dosage of oral administration, and further improve the safety of oral administration of the compound.
Detailed Description
The following examples will allow one skilled in the art to more fully understand the invention, but are not intended to limit the invention in any way, the structure of all compounds being determined by MS or 1 H-NMR, all optical isomers of the single configuration involved being configuration determined by optical rotation testing or electron circular dichroism.
In this example, unless otherwise specified, the solvents and reagents used were all commercially available. The starting materials were all commercially available.
Embodiment one: synthesis of M11
Synthesis of Compound 2:
Compound 1 (100 g,793mmol,1 eq) was dissolved in DMF (1.5L), potassium carbonate (219 g,1.59mol,2 eq) was added, the ice water bath was cooled to 0℃and benzyl bromide (203 g,1.19mol,1.5 eq) was added dropwise, and after the addition was completed, the reaction was carried out at 0℃for 30 minutes, and then transferred to an oil bath pot for 5 hours at 80 ℃. TLC (EA/PE=1/2, EA is ethyl acetate, PE is petroleum ether) detects that there is only a small amount of remaining raw material, after-treatment, the system is cooled to room temperature, poured into water (3L), extracted with ethyl acetate (300 ml x 3), the organic phases are combined and washed with water (100 ml x 3), saturated brine (200 ml x 1), dried over anhydrous sodium sulfate for 20 minutes, and the organic phase is filtered, concentrated and purified by column chromatography (EA/PE=1/5 to 1/1) to obtain 157g of product. Yellow oil, yield 91.8%.
Synthesis of Compound 3:
Compound 2 (100 g,115mmol,1 eq) was dissolved in bromobenzene (1L) at room temperature and reacted with SeO 2 (152 g,347mmol,3 eq) in an oil bath at 180deg.C for 16 hours, and the reaction was complete as determined by TLC (EA/PE=1/2). The system was cooled to room temperature, celite was added to a buchner funnel and filtered, washed with dichloromethane celite (100 ml x 3), the organic phase was combined and concentrated in dichloromethane with water pump and bromobenzene was concentrated in oil pump to give crude product (100 g, red oil).
Synthesis of Compound 4:
Compound 3 (100 g, 433 mmol,1 eq) was dissolved in DMSO (1.5L) at room temperature, cyclopropylecarbaldehyde (91 g,1.3mol,3 eq) was added, the temperature was reduced to 0℃in an ice-water bath, pyrrolidine (31 g,434mmol,1 eq) was added dropwise under nitrogen protection, the reaction was continued for 30 minutes at 15℃after the dropwise addition, and the mixture was transferred to an oil bath pot at 50℃for 5 hours. TLC (DCM/MeOH=20/1) checked complete reaction of starting material, cooled the system to room temperature, poured into water (3L), extracted with ethyl acetate (250 ml x 3), combined organic phases washed with water (100 ml x 3), saturated brine (100 ml x 1), dried over anhydrous sodium sulfate for 30min, and concentrated column chromatography purification of the organic phase (EA/PE=1/10-1/1) gave the product (32 g, red oil, yield 24.6％).ESI-MS(+):m/z＝301.1;1H NMR(CDCl3,400MHz)：δ8.85(s,1H),7.69-7.67(d,1H),7.28-7.43(m,5H),6.42-6.43(d,1H),5.28(s,2H),4.93-4.95(d,1H),2.79-2.81(d,1H),1.15-1.27(m,4H).
Synthesis of Compound 5:
Compound 4 (32 g,107mmol,1 eq) was dissolved in methanol (300 ml)/water (150 ml) under ice-water bath, trifluoroacetyl hydrazine (27.3 g,214mmol,2 eq) was added and reacted in ice-water bath for 30 min after the addition, oil bath 50℃for 4 hours, TLC (DCM/MeOH=20/1) detected complete reaction of starting material. The reaction system was cooled to room temperature, methanol in the system was concentrated, water (150 ml) was added, extraction was performed with ethyl acetate (100 ml x 3), saturated brine (500 ml x 1) was combined with the organic phase, dried over anhydrous sodium sulfate for 10 minutes, and the organic phase was filtered, concentrated, column-chromatographed (EA/pe=1/10 to 1/1) to give a product (14 g, yellow solid, yield 45％).ESI-MS(+):m/z＝297.1;1H NMR(DMSO-D6,400MHz)：δ7.90-7.92(d,1H),7.31-7.43(m,5H),7.14-7.15(s,1H),6.28-6.30(d,1H),5.78-5.79(d,1H),5.27-5.30(d,1H),5.03-5.06(d,1H),4.09-4.10(m,1H),1.19-1.28(m,2H),0.85-0.90(m,1H),0.61-0.64(m,1H).
Compound 6 synthesis:
Compound 5 (14 g,47.2mmol,1 eq) was dissolved in THF (150 ml) under an ice-water bath and triethylamine (9.6 g,94.5mmol,2 eq) and DMAP (1.73 g,14.2mmol,0.3 eq) were added. Acetic anhydride (9.6 g,94.5mmol,2 eq) was added dropwise at 0℃and after the addition was completed, the reaction was continued at 0℃for 30 minutes and then allowed to proceed to room temperature for 2 hours. TLC (EA/pe=1/1) checked the completion of the starting material reaction. The system was poured into water (150 ml), extracted with ethyl acetate (50 ml x 3), the combined organic phases were saturated brine (100 ml x 2), dried over anhydrous sodium sulfate for 10 min, and the organic phases were purified by filtration, concentration, column chromatography (EA/pe=1/10 to 1/1) to give product 6 (15 g, yellow solid, yield 94%).
Compound 7 synthesis:
Compound 6 (15 g,44.3mmol,1 eq) was dissolved in THF (150 ml) and methanol (50 ml) under ice-water bath, sodium borohydride (3.35 g,133mmol,3 eq) was added in portions at 0℃and the reaction was carried out for 30 minutes at 0℃after the addition was completed, and the reaction was allowed to proceed to room temperature for 2 hours. TLC (EA/PE=1/1) checked for disappearance of starting material, the system was poured into an aqueous solution of ammonium chloride (150 ml), extracted with ethyl acetate (50 ml x 3), the combined organic phases saturated brine (50 ml x 3), dried over anhydrous sodium sulfate for 10min, the organic phases filtered and concentrated to give the crude product (15 g, yellow solid) which was taken directly to the next step without purification.
Compound 8 synthesis:
Compound 7 (15 g,44mmol,1 eq) was dissolved in THF (150 ml), DMAP (0.54 g,4.4mmol,0.1 eq), triethylamine (8.92 g,88mmol,2 eq) were added dropwise at 0℃and Boc 2 O (19.2 g,88mmol,2 eq) was added and reacted for 30 minutes at 0℃after the addition was completed and then allowed to react for 2 hours at room temperature. TLC (DCM/MeOH=20/1) checked for disappearance of starting material, poured the system into water (150 ml), extracted with ethyl acetate (50 ml x 3), combined with saturated brine (50 ml x 3), dried over anhydrous sodium sulfate for 10min, and concentrated by filtration and column chromatography purification of the organic phase (EA/PE=1/10-1/1) gave the product (15 g, yellow solid, yield 78%).
Compound 9 synthesis:
Compound 8 (15 g,34mmol,1 eq) was dissolved in methanol (150 ml) under an ice-water bath, and potassium carbonate (4.7 g,34mmol,1 eq) was added in portions and reacted at 0℃for 30 minutes at room temperature for 6 hours. TLC (DCM/meoh=20/1) checked complete reaction of the starting materials, poured the system into water (150 ml), extracted with ethyl acetate (50 ml x 3), combined with saturated brine (30 ml x 3), dried over anhydrous sodium sulfate for 10min and the organic phase filtered and concentrated to give crude product (10 g, yellow solid).
Compound 10 synthesis:
Compound 9 (10 g,25mmol,1 eq) was dissolved in DCM (100 ml) under ice-water bath and Dess-Martin oxidant (16 g,37.6mmol,1.5 eq) was added in portions at 0deg.C and reacted at 0deg.C for 30 min at room temperature for 5 hours. TLC (DCM/meoh=15/1) checked complete reaction of starting material, the system was poured into aqueous sodium bicarbonate (100 ml), DCM extracted (30 ml x 3) and the organic phase was combined with saturated brine (30 ml x 1). Drying with anhydrous sodium sulfate for 10 min, filtering, concentrating, column chromatography purifying (EA/PE=1/10-1/0) the organic phase to obtain crude product 10g, pulping the crude product with ethyl acetate to obtain pure product 5.4g, concentrating the mother liquor to obtain crude product 4.6g.ESI-MS(+):m/z＝397.3;1H NMR(DMSO-D6,400MHz)：δ7.84-7.86(d,1H),7.28-7.46(m,5H),6.30-6.32(d,1H),5.07-5.20(m,2H),3.85-4.20(dd,2H),1.43(s,9H),1.35-1.37(m,1H),1.18-1.28(m,2H),1.08-1.12(m,1H).
Synthesis of Compound 12
After nitrogen is replaced in the three-mouth bottle, 226ml of phenylmagnesium bromide (2.8 mol/L) is added, the temperature is reduced to below 10 ℃ under ice water bath, 49.9g of selenium powder is added in batches, the reaction temperature is controlled to be not more than 30 ℃, and the reaction is carried out for 2 hours after the selenium powder is added. Adding 2mol/L hydrochloric acid under ice-water bath, reacting to release heat obviously, adding EA for extraction, separating liquid, and spin-drying organic phase to obtain the product 12 as brown oily substance with strong odor. The next reaction was directly carried out.
Synthesis of Compound 14
After nitrogen is replaced in the three-port bottle, 81.18g of LDA (lithium diisopropylamide) is added, the temperature is reduced to minus 30 ℃ under dry ice-ethanol solution, 50g of THF solution of 3, 4-difluorobenzoic acid is dropwise added, the heat release is obvious in the dropwise adding process, the temperature of a reaction system is controlled to be lower than minus 20 ℃, the reaction is carried out for 2 hours, DMF is added, the heat release is obvious, the temperature of the reaction system is controlled to be lower than minus 20 ℃, the temperature is not required to be controlled, the reaction is gradually carried out to room temperature, the reaction is carried out overnight, and the sampling detection reaction is finished. HCl is added into the reaction system, the heat release is obvious, EA is added for extraction, the solution is separated, the organic phase is dried by spinning, 72g of crude product of the compound 14 is obtained, the obtained product is directly used for the next reaction, the obtained crude product is yellow solid, and the yield is 122% (DMF in the material is not dried by spinning).
Synthesis of Compound 15
After nitrogen was replaced in the three-port flask, 58.81g of compound 12, 49.6g of compound 14, 300ml of toluene, 17.6g of camphorsulfonic acid were added, and the temperature was raised to 70℃and the reaction was carried out overnight. Cooling the reaction system to room temperature, adding NaOH solution, separating liquid, adding EA (ethylene oxide) into water phase for extraction, combining organic phases, washing the organic phases with saturated NaCl, and spin-drying to obtain a crude product: 123.08g, pulping the obtained crude product by PE, and carrying out suction filtration to obtain 34.4g of product, and recovering 14.83g of pulping filtrate. Compound 15 was an orange solid in 47.9% yield.
Synthesis of Compound 16
16.9G of AlCl 3 and 250ml of toluene are added into a three-necked flask, the system is cooled in an ice water bath, 17.1g of tetramethyl disiloxane is added, the mixture is stirred uniformly, 150ml of toluene solution of compound 15 (34.4 g) is added, the reaction is slightly exothermic, alCl 3 is gradually dissolved, and the temperature is heated to 80 ℃ for 1 hour; stopping the reaction, adding sulfuric acid solution (16.2mL+240 mL of water), separating, extracting the water phase with EA, spin-drying the organic phase, pulping the crude product with PE, and suction-filtering to obtain 22g of solid, wherein the obtained solid is yellow powdery solid with the theoretical amount of the product: 34.68g, yield 22g total of solid product compound 16: 63.4%. Directly used in the next reaction.
Synthesis of Compound 17:
Adding 429g of polyphosphoric acid into a three-mouth bottle, heating to 80 ℃, adding 42g of compound 16, heating to 120 ℃, changing the color of a reaction system from yellow to dark purple, reacting for 1 hour, sampling, adding water, and treating by EA; reducing the reaction temperature to below 100 ℃, adding water, stirring uniformly, adding EA for extraction, separating an organic phase, spin-drying, pulping a crude product by PE, filtering to obtain 3.4g of a product, separating filtrate by column chromatography to obtain 10g of the product, wherein the compound 17 is colorless to pale yellow flocculent solid, and the theoretical yield is as follows: 39.69g, actual yield: 13.4g, yield: 33.76%.
Synthesis of Compound 18:
2.12g of Compound 17 and 20ml of ethanol were added to a three-necked flask, and the system was stirred. Dropwise adding an ethanol solution of sodium borohydride (0.26 g) under an ice water bath, slightly heating a reaction system, transferring to room temperature for reaction after dropwise adding, gradually dissolving materials, sampling and detecting after the reaction solution is dissolved, and processing after the reaction is finished; adding 2mol/L hydrochloric acid until no bubbles are generated, wherein the pH value is=4-6, a large amount of solids are separated out, filtering to obtain solids, extracting filtrate with EA, separating an organic phase, and spin-drying the organic phase to obtain a product 18 which is brown solid and directly used for the next reaction.
Synthesis of Compound 19
2.5G of compound 10, 2.13g of compound 18, 5.44g of compound T3P (propylphosphoric anhydride), 1.1g of methanesulfonic acid and 30ml of EA are added into a single-port bottle, the mixture is heated to reflux, reacted overnight, sampled and detected, and the mixture is treated after the reaction is completed; post-treatment: saturated aqueous sodium bicarbonate was added until no bubbles evolved, the solution was separated, the aqueous phase was extracted with EA, the organic phases were combined, dried by spinning, and the crude product was slurried with PE to give product 19 as a pale brown powdery solid, theoretical yield: 3.36g, actual yield: 2.2g, yield: 65.48%.
Synthesis of Compound M01:
2.6g of compound 19, 0.94g of compound LiCl and 20ml of DMA (N, N-dimethylacetamide) are added into a single-port bottle, the temperature is raised to 100 ℃, the reaction liquid is yellow and turbid, the reaction is carried out for 2 hours, sampling detection is carried out, and the treatment is carried out after the reaction is completed; adding saturated sodium bicarbonate aqueous solution, separating out solid, filtering, extracting filtrate with EA, spin-drying organic phase, pulping crude product with PE to obtain 2.28g of compound M01, wherein the product is brown solid powder, and the yield is: 104% (yield slightly higher than theoretical yield, judged to be solvent DMA residual). Directly used in the next reaction. ESI-MS (+): m/z=501.1.
Synthesis of compound M11:
To a single vial was added 1.1g of compound M01, 0.61g of K 2CO3, 0.55g of KI, and 0.55g of chloromethyl dimethyl carbonate. And adding 20mL of DMA (direct memory access) into the system, heating to 60 ℃, reacting overnight, sampling and detecting, and processing after the reaction is finished. Cooling the reaction system to room temperature, adding 2N HCl, adding water, precipitating solid, suction filtering, extracting filtrate with EA, separating organic phase, dissolving filter cake with EA, mixing with organic phase, drying organic phase with anhydrous sodium sulfate, spin-drying, separating by column chromatography to obtain brown solid powder 0.766g of compound M11, yield ：58.9％.ESI-MS(+):m/z＝589.5;1HNMR(DMSO-D6,400MHz)：δ7.26-7.41(m,5H),7.10-7.13(m,1H),6.92-6.98(m,2H),5.90-5.92(d,1H),5.74-5.86(d,1H),5.56-5.58(d,1H),5.38-5.42(m,2H),4.12-4.16(m,2H),4.01-4.07(m,1H),3.74(s,3H),2.79(s,1H),1.73-1.75(m,1H),1.16-1.24(m,2H),0.87-0.90(m,1H),0.72-0.74(m,1H).
Embodiment two: resolution of M19
The compound M11 is prepared and separated to obtain M19 and enantiomer M19-1, which are white solids. The preparation conditions are as follows: using a supercritical fluid chromatograph; chromatographic column: CHIRALPAK IB-N4.6 x 100mm,3 μm; the solvent is methanol, and the carrier is liquid CO 2; the pressure was 1500psi, the flow rate was set at 2.0ml/min, the column temperature 25℃and the elution was gradient. Wherein the retention time of M19 is 2.86min, and the result shows that the optical rotation is performed by taking methanol as a solvent. The retention time of M19-1 was 2.55min, and the result was shown as a D-isomer by optical rotation measurement using methanol as a solvent. The absolute configuration of M19 and M19-1 is determined by adopting a method for determining the absolute configuration of chiral compounds widely adopted internationally at present, quantitatively calculating a predicted theoretical electron circular dichroism (ECD, namely commonly referred to as circular dichroism) spectrum, comparing the predicted theoretical Electron Circular Dichroism (ECD) spectrum with an experimental ECD spectrum, and finally determining the absolute configuration by conforming an experimental ECD signal to a theoretical calculation result, wherein M19 is an S configuration and M19-1 is an R configuration.
Embodiment III: synthesis of M02
The compound M01 is prepared and separated to obtain M02 and enantiomer, which are white solids. The preparation conditions are as follows: using a supercritical fluid chromatograph; chromatographic column: CHIRALPAK IB-N4.6 x 100mm,3 μm; the solvent is methanol, and the carrier is liquid CO 2; the pressure was 1500psi, the flow rate was set at 2.0ml/min, the column temperature 25℃and the elution was gradient. Optical rotation was performed using methanol as solvent, M02 as the levorotatory form and M02 enantiomer as the dextrorotatory form. As can be seen from comparison with M19 and M19-1, the absolute configuration of M02 is the S configuration.
Embodiment four: synthesis of M03 and M04
Synthesis of Compound 23
Under the protection of nitrogen, 1g of compound 17 is dissolved in 15ml of THF, 140mg of lithium aluminum hydride-D4 is slowly added at 0 ℃, and the system is heated to 25 ℃ to react for 8 hours. The system is cooled to 0 ℃, and water is added for quenching reaction. The system was extracted with 2N hydrochloric acid and EA. The organic phase was concentrated to dryness. Column chromatography gave compound 23 in a total of 0.58g, 57% yield, ESI-MS (+): m/z=314.0.
Synthesis of Compound 24
Referring to the synthetic method for compound 19, 0.94g of compound 24 was synthesized, ESI-MS (+): m/z= 592.1.
Synthesis of Compound M03
Referring to the synthesis of compound M01, 0.51g of compound M03 was synthesized, ESI-MS (+): M/z=502.1.
Synthesis of Compound M04
Referring to the preparation and separation method of the compound M01, the compound M04 is obtained, and the M04 is still a levorotatory body and the absolute configuration is an S configuration.
Fifth embodiment: synthesis of M05 and M06
Synthesis of Compound 25:
Under the protection of nitrogen, 1g of compound 17 is dissolved in 15ml of THF, 4ml of methyl lithium reagent (1.6M diethyl ether solution) is slowly added dropwise at-20 ℃, and the system is naturally warmed to 25 ℃ for reaction for 8 hours. The system is cooled to 0 ℃, and water is added for quenching reaction. Concentrating to dryness, and extracting with EA and water. The organic phase was separated and concentrated to dryness. Column chromatography gave compound 25 in a total of 0.77g, 73% yield, ESI-MS (+):m/z=327.1.
Synthesis of Compound 26
Referring to the synthesis of compound 19, compound 26 amounted to 0.94g, ESI-MS (+): m/z=605.2 was synthesized.
Synthesis of Compound M05
Referring to the synthesis of compound M01, 0.51g of compound M05 was synthesized, ESI-MS (+): M/z=515.1.
Synthesis of Compound M06
Referring to the preparation and separation method of the compound M01, the compound M06 is obtained, and the M06 is confirmed to be still a levorotatory body, and the absolute configuration is an S configuration.
Example six: synthesis of B-1
Synthesis of Compound B-1:
100mg of compound M02, 130mg of DIPEA (diisopropylethylamine), 73mg of DMAP (4-dimethylaminopyridine) and 2ml of anhydrous methylene chloride were added to the reaction flask at room temperature, followed by 104mg of diethyl chlorophosphate. The reaction was carried out at room temperature overnight, dichloromethane and water were added, the organic phase was separated by extraction, dried over anhydrous sodium sulfate, and the organic phase was concentrated to dryness, and purified by column chromatography to give 77.6mg of Compound B-1 in 61% yield, ESI-MS (+): m/z= 637.1.
Embodiment seven: synthesis of Compound B-5:
Synthesis of Compound 30
Compound 30 was synthesized by the synthetic method described in reference (chem. Eur. J.2011,17, 1649-1659).
A solution of 4.45g of phosphorus oxychloride in THF (35 ml) was cooled to-78℃and then a solution of 3g of compound 29 and 3.14g of triethylamine in THF (tetrahydrofuran, 45 ml) was added dropwise to the system. After the dripping is finished, the system is heated to-50 ℃ to react for 1h, then heated to 25 ℃ to react for 4h, the system is filtered to remove triethylamine salt, and then 5ml of water is dripped into the solution to react for 2h. The system was concentrated to dryness and purified by column chromatography to give 3.06g of compound 30 in 68% yield, ESI-MS (+): m/z=187.0.
Synthesis of Compound 31
Compound 31 was synthesized by the method described in the literature (Journal of MEDICINAL CHEMISTRY,2020, vol.63, #24, p.15785-15801).
Compound 30 was dissolved in water at 0deg.C, tetrabutylammonium bisulfate and sodium bicarbonate were added, and after 15 minutes of reaction, a solution of chloromethyl chlorosulfonate in DCM was added to the system. The system was reacted at 25℃for 18h. The organic phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to dryness, and then subjected to column chromatography to obtain 0.6g of Compound 31 in 16% yield.
Synthesis of Compound B-5
The synthesis of reference compound B-1 gave 160mg of compound B-5 in 54% yield, ESI-MS (+): m/z=699.0.
Example eight: synthesis of B-6, L-1 and L-2
Synthesis of Compound B-6
Under the protection of nitrogen, 100mg of compound M02 and 10mL of dry acetonitrile are added into a reaction bottle for stirring and dissolution, the mixture is cooled to-25 ℃, 60mg of dry carbon tetrachloride, 52mg of diisopropylethylamine and 2.5mg of 4-dimethylaminopyridine are sequentially added, 58mg of dibenzyl phosphite and 5mL of dry acetonitrile mixed solution are slowly added dropwise, the temperature is controlled below-10 ℃, the reaction is continued for 3.5h after the dropwise addition, the temperature is naturally raised to 20-25 ℃, 0.5mol/L of monopotassium phosphate is added for stopping the reaction, the ethyl acetate is used for extraction, the organic layers are combined, the mixture is sequentially washed by distilled water and saline, the organic phases are concentrated to dryness, and the column chromatography is carried out to obtain 97.35mg of compound B-6, the yield is 64%, ESI-MS is (+): M/z= 761.1.
Synthesis of Compounds L-1 and L-2
40Mg of compound B-6, 16mg of sodium iodide and 2ml of anhydrous acetonitrile are added into a reaction bottle, the system is heated to 82 ℃ for reaction for 3 hours, solids are separated out, the system is filtered by suction to obtain 30mg of solid of compound L-1, the yield is 85%, and ESI-MS (-):m/z= 669.1. Compound L-1 was dissolved with water, added to an acidic ion exchange resin, stirred for 10 minutes, suction filtered, and concentrated to dryness to give compound L-2, esi-MS (-): m/z= 669.1.
Example nine: synthesis of B-7
To a single vial was added 70mg of compound M02, 91.34mg of Cs 2CO3, 28mg of KI, 224mg of dibenzyl chloromethyl phosphate, and 2mL of DMA. The temperature of the system is raised to 50 ℃, the reaction is carried out for 2.5h, and the reaction is completed. The reaction system is cooled to room temperature, EA and water are used for extraction, an organic phase is separated, the organic phase is dried by anhydrous sodium sulfate and then spin-dried, and the solid 68.6mg of the compound B-7 is obtained after column chromatography separation, and the yield is: 62%. ESI-MS (+): m/z= 791.2
Example ten: synthesis of L-3
To a single vial was added 70mg of compound M02, 91.34mg of Cs 2CO3, 28mg of KI, 224mg of dibenzyl chloromethyl phosphate, and 2mL of DMA. The temperature of the system is raised to 50 ℃, the reaction is carried out for 2.5h, and the reaction is completed. 1ml of methanol is added into the reaction system and stirred for 10 minutes, the methanol and the DMA reaction liquid are evaporated, and 60mg of monobenzyl ester compound L-3 is obtained through c18 column chromatography, and the yield is: 50%. ESI-MS (-) m/z= 699.1 example eleven: synthesis of L-3
The compound L-5-0 is synthesized according to the method of patent (CN 110300753B) and is prepared and separated by chiral column. Referring to the synthesis of compound L-3, compound L-5, ESI-MS (-): m/z=651.1 was obtained.
Comparative example one: synthesis of DB-1
According to the synthesis method of the reference compound M19, L-5-0 is used as a material to synthesize the compound DB-1 in one step. DB-1 through optical rotation test and with M19 optical rotation comparison, confirm DB-1 is the left-handed body, S configuration.
The following examples were synthesized in the same manner as in the above examples, using as the starting materials the commercially available compounds or intermediate compounds appropriately synthesized from the commercially available compounds or other intermediates synthesized in the patent of the present invention:
Or a hydrate, solvate, optical isomer, polymorph, isotopic derivative, pharmaceutically acceptable salt thereof.
Embodiment twelve: solubility test in Water
The solubility of the compound of the present invention in water was tested according to the chinese pharmacopoeia 2020 edition guide to the use solubility determination method: 0.1000g of the sample to be ground into fine powder is weighed, added into a certain amount of water at 25+/-2 ℃ and shaken vigorously for 30 seconds every 5 minutes, and the dissolution condition within 30 minutes is observed, and if no solute particles are visible, the complete dissolution is considered. The results are shown in Table 1:
Table 1: results of the solution test
Compounds of formula (I) Dissolution status Compounds of formula (I) Dissolution status Compounds of formula (I) Dissolution status
B-1 Slightly soluble B-3 Slightly soluble B-5 Slightly soluble
L-1 Dissolving L-2 Dissolving L-3 Dissolving
L-4 Dissolving L-5 Dissolving L-7 Dissolving
L-8 Dissolving M-19** Hardly soluble DB-1* Hardly soluble
* Compound M-19 was used as a control compound;
* Compound DB-1 is compound 45 of patent CN 110300753B, used as a comparative compound;
the test results show that: the compounds of the comparative examples were hardly dissolved, but the solubility of the compounds of the present invention was significantly improved, and in particular, the water-solubility of the compounds L-1 to L-8 was better. Solubility is a key factor affecting drug availability, while an increase in solubility more facilitates dissolution of drug molecules in the gastrointestinal tract; meanwhile, the solubility is obviously improved, so that the method is suitable for developing true solution preparations.
Embodiment thirteen: stability study
Preparation of simulated gastric fluid: precisely measuring 4.5ml of 36% hydrochloric acid into a 1L volumetric flask, adding water to the scale, shaking for later use, and marking as stock solution. Precisely weighing the stock solution in a volumetric flask with volume of 10ml to 50ml, precisely weighing 500.0mg pepsin into the volumetric flask, adding water to scale, performing ultrasonic treatment until dissolution, filtering to obtain a clear solution, and marking as simulated gastric juice.
1.0Mg of the sample is precisely weighed into a 5ml volumetric flask, 2.5ml of isopropanol is firstly added into the volumetric flask, the solution is shaken and dissolved, and then simulated gastric fluid (pH 2.0) is added to the scale. Shaking and shaking evenly, and filtering to simulate the stability investigation of gastric juice.
1.0Mg of the sample is precisely weighed into a 5ml volumetric flask respectively, 2.5ml of isopropanol is firstly added into the volumetric flask, the solution is shaken for dissolution, and then water is added to the scale. Shaking and shaking evenly, and filtering for solution stability investigation.
Stability investigation after the sample was sealed from light, the sample was left at 25 ℃ ± 2 ℃ for 6 hours, and the solution stability and stability in simulated gastric fluid of the sample were checked by HPLC, and the results are shown in table 2:
table 2: solution stability of sample and stability in simulated gastric fluid
The results show that: the compound has good stability in the state of solution and simulated gastric fluid, and can meet the requirement of drug administration.
Fourteen examples: cytopathic degree (CPE) assay
MDCK cells are inoculated into 96-well culture plates and cultured at 37 ℃ with 5% CO 2. During the cell index growth period, maintaining liquid containing different dilutions of sample and positive reference medicine is added, and 3 compound wells are set in each concentration, and normal cell reference wells are set simultaneously. After the sample was added, the sample was cultured for 72 hours, and the cytotoxicity test of the sample was performed by the CPE method. MDCK cells were also inoculated into 96-well plates and incubated at 37 ℃ with 5% co 2. After 24 hours, influenza virus (A/han-defenses/359/95 (H3N 2)) was infected, adsorbed for 2 hours, the virus solution was discarded, a maintenance solution containing samples of different dilutions and positive control was added, 3 duplicate wells were set for each concentration, and cell control wells and virus control wells were set simultaneously, 5% CO 2 was used, and incubated at 37 ℃. The antiviral test of the tested sample is carried out by a CPE method, and when the pathological change degree (CPE) of the virus control group reaches 4+, the cytopathic change degree (CPE) of each group is observed. The half-toxic concentration of the sample on cells (TC 50) and the drug effective concentration (EC 50) to suppress 50% of the cytopathic effect were calculated by the Reed-Muench method, respectively, and the Therapeutic Index (TI) was calculated as TC 50/EC50.
Table 3: anti-influenza virus cell activity/toxicity data
Experiments show that the compound has better safety and anti-influenza virus activity, and compared with the positive control Ballo Sha Weiji Ballo Sha Weizhi, the compound has better anti-influenza virus cell activity.
Example fifteen: oral pharmacokinetic test in rats
The jugular vein of 12 SD rats, male, 180g-220g, was cannulated before the start of the experiment, and the experiment was started after three days (free feeding of drinking water, room temperature: 20-26 ℃ C.; humidity: 40-70%; light illumination: dark = 12h:12 h). The experimental animals were divided into 4 groups of 3 animals each. The suspensions of the test substances (test substances B-3, L-3, M19 and DB-1 were each suspended with 0.5% sodium carboxymethylcellulose) were administered orally by gavage, respectively, at a dose of 2.25mg/kg in terms of M19, and equimolar. Fasted food begins at 5 pm on the day before administration but water is not forbidden, and fasted for 16-17h. Animals fed after 4h of administration, and water is not forbidden in the whole process.
15Min, 30min, 1h, 2h, 4h, 6h, 8h, 10h, 24h before and after administration. About 0.25mL of whole blood was taken through the jugular vein in heparin sodium anticoagulants, respectively. After taking the blood, the anticoagulation tube containing the blood sample is immediately reversed for 5-10 times and temporarily stored in an ice bath. The blood samples were centrifuged at 3000rpm at 4℃for 5 minutes within 1 hour after collection. Transferring the centrifugally collected plasma into a new labeled centrifuge tube, temporarily storing in a refrigerator at-20 ℃, and after all samples are collected, delivering the collected plasma to a biological sample manager and storing in the refrigerator at-80 ℃. After the biological sample is treated, the detection of the detection object is carried out through LC-MS/MS (the detection objects of groups B-3, L-3 and M19 are M02, and the detection object of group DB-1 is L-5-0). The essential pharmacokinetic parameters were calculated according to the non-compartmental model method using WinNonlin 7.0. The main pharmacokinetic parameters after intragastric administration of each set of samples are shown in table 4:
Table 4: main pharmacokinetic parameters of test sample in rat after equimolar gastric lavage administration
Parameters (parameters) T1/2 Tmax Cmax AUC0-∞
Unit (B) h h ng/ml h*ng/ml
B-3 3.89 1 38.8 347.236
L-3 4.13 0.5 43.1 373.641
M19 4.33 1 34.5 289.914
DB-1 4.73 0.5 20.3 212.359
The results of pharmacokinetic studies on intragastric administration in rats showed that: the in vivo exposure (AUC) of the compounds of the invention was significantly increased after oral gavage administration in rats compared to M19 and DB-1, indicating a higher bioavailability of the compounds of the invention.
Example sixteen: cynomolgus monkey pharmacokinetic study
12 Cynomolgus monkeys, male, 3-6kg, were randomly divided into 4 groups (A/B/C/D groups) after 3 days of adaptive feeding. Group A was intravenously injected with 0.2mg/kg of a physiological saline solution of Compound L-2, and group B was orally administered with 2mg/kg of a physiological saline solution of Compound L-2 by gavage; group C was intravenously injected with 0.2mg/kg of a physiological saline solution of Compound L-4, and group D was orally administered with 2mg/kg of a physiological saline solution of Compound L-4 by gavage; during the experiment, the intravenous injection group animals can eat and drink water freely, and the oral gavage group animals can fast for more than 12 hours but not forbidden water before administration.
For intravenous administration groups, about 0.5mL of whole blood was taken through the jugular vein before and after administration for 0.033, 0.083, 0.17, 0.25, 0.5, 1, 2, 4, 8, 24 hours, respectively; for the oral gavage administration group, about 0.5mL of whole blood is taken through the jugular vein before and after 15min, 30min, 1h, 2h, 4h, 6h, 8h, 10h and 24h respectively; the whole blood sample is placed in a heparin sodium anticoagulation tube, immediately inverted for 5-10 times and temporarily stored in an ice bath. The blood samples were centrifuged at 3000rpm at 4℃for 5 minutes within 1 hour after collection. Transferring the centrifugally collected plasma into a new labeled centrifuge tube, temporarily storing in a refrigerator at-20 ℃, and after all samples are collected, delivering the collected plasma to a biological sample manager and storing in the refrigerator at-80 ℃. The biological sample was treated and then tested for compound M02 by LC-MS/MS. The principal pharmacokinetic parameters were calculated using WinNonlin 7.0 according to the non-compartmental model method and the bioavailability was calculated and the results are shown in table 5:
Table 5: pharmacokinetic parameters for cynomolgus monkey drug administration
Parameters (parameters) T1/2 Tmax Cmax AUC0-∞ F
Unit (B) h h ng/ml h*ng/ml ％
Group L-2 (IV) 11.55h / / 174 /
L-2 (PO) group 11.13 2.0 101 955 54.88％
Group L-4 (IV) 11.08h / / 156 /
L-4 (PO) group 10.87 2.0 93 891 57.11％
As can be seen from the table, when the cynomolgus monkey is orally taken and injected, the compound L-2 and the compound L-4 have better in-vivo exposure, and the absolute bioavailability reaches 54.88% and 57.11% respectively. This demonstrates that the compounds of the present invention can be administered by injection and orally at the same time, and have high oral bioavailability.
The present application has been described in terms of several embodiments, but the description is intended to be illustrative and not limiting and many more embodiments and implementations are possible within the scope of the described embodiments.

Claims (10)

1. A phosphate compound represented by formula (I), or a hydrate, solvate, optical isomer, polymorph, isotopic derivative, pharmaceutically acceptable salt thereof:
in formula (I), R a is selected from hydrogen or deuterium or methyl;
R b and R c are each independently selected from hydrogen, deuterium, or methyl, or R b and R c together with the attached carbon form cyclopropyl;
X 1 is an O atom or an S atom;
x 2 is a Se atom or an S atom;
n1 is 0, 1 or 2;
each R 1 or R 2 is independently selected from hydrogen or methyl;
R 3 and R 4 are each independently selected from the following groups: hydroxy, C1-C8 alkoxy, C3-C8 cycloalkoxy, C3-C8 heterocycloalkoxy, C6-C10 aryloxy, C7-C12 aralkyloxy, and R 3 and R 4 must not be both C1-C8 alkoxy when X 2 is S atom; or R 3 and R 4 together with the phosphorus atom to which they are attached, e.g A 5-7 membered ring of (2); wherein R 5、R6,R7、R8、R9、R10、R11、R12 and R 13 are each independently hydrogen or C1-C3 alkyl, or R 5 and R 6,R7 and R 8,R10 and R 11,R11 and R 12 each together with the carbon atom to which they are attached form an aromatic ring.
2. The phosphate compound according to claim 1, which is represented by formula (ii):
The substituents in formula (II) are as defined in claim 1.
3. The phosphate compound according to claim 1 or 2, which is represented by formula (iii):
The substituents in formula (III) are as defined in claim 1.
4. The phosphate compound according to any one of claims 1 or 2, which is represented by formula (iv):
the substituents in formula (IV) are as defined in claim 1.
5. The phosphate ester compound according to claim 3, wherein the phosphate ester compound is selected from the group consisting of sodium salt, potassium salt, magnesium salt, zinc salt, amine salt, and basic amino acid salt.
6. The phosphate compound according to any one of claims 1 to 5, which is selected from the following structures:
Or a hydrate, solvate, optical isomer, polymorph, isotopic derivative, pharmaceutically acceptable salt thereof.
7. A pharmaceutical composition comprising a compound of any one of claims 1-6, or a hydrate, solvate, optical isomer, polymorph, isotopic derivative, pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier thereof.
8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition is in the form of a tablet, capsule, powder, granule, pill, suspension, syrup, or injection.
9. The compound of any one of claims 1-6, including hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof, or the pharmaceutical composition of claim 7, for use against influenza virus.
10. Use of a compound according to any one of claims 1-6, including hydrates, solvates, optical isomers, polymorphs, isotopic derivatives, pharmaceutically acceptable salts thereof, or a pharmaceutical composition according to claim 7 for the preparation of an anti-influenza virus medicament.
CN202211395789.6A 2022-11-09 2022-11-09 Anti-influenza virus phosphate compound and application thereof Pending CN118047811A (en)

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