WO2024061048A1 - 一类石蒜碱衍生物、其药物组合物及其在制备抗病毒药物中的用途 - Google Patents
一类石蒜碱衍生物、其药物组合物及其在制备抗病毒药物中的用途 Download PDFInfo
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- WO2024061048A1 WO2024061048A1 PCT/CN2023/118167 CN2023118167W WO2024061048A1 WO 2024061048 A1 WO2024061048 A1 WO 2024061048A1 CN 2023118167 W CN2023118167 W CN 2023118167W WO 2024061048 A1 WO2024061048 A1 WO 2024061048A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- lycorine
- substituted
- pharmaceutically acceptable
- pharmaceutical composition
- acceptable salt
- Prior art date
Links
- XGVJWXAYKUHDOO-DANNLKNASA-N lycorine Chemical class C1CN2CC3=CC=4OCOC=4C=C3[C@H]3[C@H]2C1=C[C@H](O)[C@H]3O XGVJWXAYKUHDOO-DANNLKNASA-N 0.000 title claims abstract description 46
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000003443 antiviral agent Substances 0.000 title claims description 6
- 150000003839 salts Chemical class 0.000 claims abstract description 14
- 150000001875 compounds Chemical class 0.000 claims description 30
- XGVJWXAYKUHDOO-UHFFFAOYSA-N galanthidine Natural products C1CN2CC3=CC=4OCOC=4C=C3C3C2C1=CC(O)C3O XGVJWXAYKUHDOO-UHFFFAOYSA-N 0.000 claims description 21
- KQAOMBGKIWRWNA-UHFFFAOYSA-N lycorine Natural products OC1C=C2CCN3C2C(C1O)c4cc5OCOc5cc34 KQAOMBGKIWRWNA-UHFFFAOYSA-N 0.000 claims description 21
- -1 phenoxy, phenylmercapto, substituted phenoxy, substituted phenylmercapto Chemical class 0.000 claims description 18
- 238000006243 chemical reaction Methods 0.000 claims description 15
- 241000700605 Viruses Species 0.000 claims description 13
- 125000001424 substituent group Chemical group 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 9
- 125000006273 (C1-C3) alkyl group Chemical group 0.000 claims description 8
- 229910052736 halogen Inorganic materials 0.000 claims description 8
- 150000002367 halogens Chemical class 0.000 claims description 8
- 150000001924 cycloalkanes Chemical class 0.000 claims description 5
- 229910052739 hydrogen Inorganic materials 0.000 claims description 5
- 239000001257 hydrogen Substances 0.000 claims description 5
- 238000006467 substitution reaction Methods 0.000 claims description 5
- 125000006559 (C1-C3) alkylamino group Chemical group 0.000 claims description 4
- 125000006274 (C1-C3)alkoxy group Chemical group 0.000 claims description 4
- 230000009471 action Effects 0.000 claims description 4
- 125000002252 acyl group Chemical group 0.000 claims description 4
- 239000002552 dosage form Substances 0.000 claims description 4
- 125000004970 halomethyl group Chemical group 0.000 claims description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 3
- 241000907316 Zika virus Species 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 150000007524 organic acids Chemical class 0.000 claims description 3
- 125000000951 phenoxy group Chemical group [H]C1=C([H])C([H])=C(O*)C([H])=C1[H] 0.000 claims description 3
- 241001429382 Coxsackievirus A16 Species 0.000 claims description 2
- 241000315672 SARS coronavirus Species 0.000 claims description 2
- 125000000217 alkyl group Chemical group 0.000 claims description 2
- 229910052794 bromium Inorganic materials 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 229910052801 chlorine Inorganic materials 0.000 claims description 2
- 239000007822 coupling agent Substances 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 229910052731 fluorine Inorganic materials 0.000 claims description 2
- 125000005842 heteroatom Chemical group 0.000 claims description 2
- 125000000592 heterocycloalkyl group Chemical group 0.000 claims description 2
- 150000002431 hydrogen Chemical class 0.000 claims description 2
- 238000002347 injection Methods 0.000 claims description 2
- 239000007924 injection Substances 0.000 claims description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 2
- 229910052717 sulfur Inorganic materials 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 241001678559 COVID-19 virus Species 0.000 claims 1
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- 238000013270 controlled release Methods 0.000 claims 1
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- 239000006187 pill Substances 0.000 claims 1
- 239000007921 spray Substances 0.000 claims 1
- 238000013268 sustained release Methods 0.000 claims 1
- 239000012730 sustained-release form Substances 0.000 claims 1
- 230000000840 anti-viral effect Effects 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 201000010099 disease Diseases 0.000 abstract description 2
- 230000003612 virological effect Effects 0.000 abstract 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 19
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 16
- 229940079593 drug Drugs 0.000 description 15
- 239000003814 drug Substances 0.000 description 15
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 14
- 241000711573 Coronaviridae Species 0.000 description 13
- 239000007787 solid Substances 0.000 description 12
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 11
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 10
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 9
- 210000003501 vero cell Anatomy 0.000 description 8
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- 230000005764 inhibitory process Effects 0.000 description 6
- RWWYLEGWBNMMLJ-MEUHYHILSA-N remdesivir Drugs C([C@@H]1[C@H]([C@@H](O)[C@@](C#N)(O1)C=1N2N=CN=C(N)C2=CC=1)O)OP(=O)(N[C@@H](C)C(=O)OCC(CC)CC)OC1=CC=CC=C1 RWWYLEGWBNMMLJ-MEUHYHILSA-N 0.000 description 6
- RWWYLEGWBNMMLJ-YSOARWBDSA-N remdesivir Chemical compound NC1=NC=NN2C1=CC=C2[C@]1([C@@H]([C@@H]([C@H](O1)CO[P@](=O)(OC1=CC=CC=C1)N[C@H](C(=O)OCC(CC)CC)C)O)O)C#N RWWYLEGWBNMMLJ-YSOARWBDSA-N 0.000 description 6
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- BCNZYOJHNLTNEZ-UHFFFAOYSA-N tert-butyldimethylsilyl chloride Chemical compound CC(C)(C)[Si](C)(C)Cl BCNZYOJHNLTNEZ-UHFFFAOYSA-N 0.000 description 5
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 4
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 4
- 208000025721 COVID-19 Diseases 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
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- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 108090001074 Nucleocapsid Proteins Proteins 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
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- 241000234270 Amaryllidaceae Species 0.000 description 2
- 241000725619 Dengue virus Species 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 241000991587 Enterovirus C Species 0.000 description 2
- 241000711549 Hepacivirus C Species 0.000 description 2
- 241000700584 Simplexvirus Species 0.000 description 2
- 108020000999 Viral RNA Proteins 0.000 description 2
- 241000710886 West Nile virus Species 0.000 description 2
- 241000710772 Yellow fever virus Species 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 2
- 210000004748 cultured cell Anatomy 0.000 description 2
- TXWOGHSRPAYOML-UHFFFAOYSA-N cyclobutanecarboxylic acid Chemical compound OC(=O)C1CCC1 TXWOGHSRPAYOML-UHFFFAOYSA-N 0.000 description 2
- NZNMSOFKMUBTKW-UHFFFAOYSA-N cyclohexanecarboxylic acid Chemical compound OC(=O)C1CCCCC1 NZNMSOFKMUBTKW-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- QWXYZCJEXYQNEI-OSZHWHEXSA-N intermediate I Chemical compound COC(=O)[C@@]1(C=O)[C@H]2CC=[N+](C\C2=C\C)CCc2c1[nH]c1ccccc21 QWXYZCJEXYQNEI-OSZHWHEXSA-N 0.000 description 2
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- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
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- 229940051021 yellow-fever virus Drugs 0.000 description 2
- KZEVSDGEBAJOTK-UHFFFAOYSA-N 1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-2-[5-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]-1,3,4-oxadiazol-2-yl]ethanone Chemical compound N1N=NC=2CN(CCC=21)C(CC=1OC(=NN=1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)=O KZEVSDGEBAJOTK-UHFFFAOYSA-N 0.000 description 1
- DCGVRKXPYLZLIS-UHFFFAOYSA-N 2-(4-fluorophenyl)sulfanylpropanoic acid Chemical compound OC(=O)C(C)SC1=CC=C(F)C=C1 DCGVRKXPYLZLIS-UHFFFAOYSA-N 0.000 description 1
- 102000012440 Acetylcholinesterase Human genes 0.000 description 1
- 108010022752 Acetylcholinesterase Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- DKHJWWRYTONYHB-UHFFFAOYSA-N CPP Chemical compound OC(=O)C(C)OC1=CC=C(Cl)C=C1 DKHJWWRYTONYHB-UHFFFAOYSA-N 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
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- TXCGAZHTZHNUAI-UHFFFAOYSA-N clofibric acid Chemical compound OC(=O)C(C)(C)OC1=CC=C(Cl)C=C1 TXCGAZHTZHNUAI-UHFFFAOYSA-N 0.000 description 1
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- JBDSSBMEKXHSJF-UHFFFAOYSA-N cyclopentanecarboxylic acid Chemical compound OC(=O)C1CCCC1 JBDSSBMEKXHSJF-UHFFFAOYSA-N 0.000 description 1
- 230000034994 death Effects 0.000 description 1
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- HTNPEHXGEKVIHG-ZJTJHKMLSA-N molnupiravir Chemical compound CC(C)C(=O)OC[C@H]1O[C@H](C(O)C1O)N1C=C\C(NC1=O)=N\O HTNPEHXGEKVIHG-ZJTJHKMLSA-N 0.000 description 1
- 229940075124 molnupiravir Drugs 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- LIENCHBZNNMNKG-OJFNHCPVSA-N nirmatrelvir Chemical compound CC1([C@@H]2[C@H]1[C@H](N(C2)C(=O)[C@H](C(C)(C)C)NC(=O)C(F)(F)F)C(=O)N[C@@H](C[C@@H]3CCNC3=O)C#N)C LIENCHBZNNMNKG-OJFNHCPVSA-N 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 229940125675 paxlovid Drugs 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
- C07D491/16—Peri-condensed 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
- C07B2200/07—Optical isomers
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- the invention belongs to the field of medical technology, and specifically relates to a class of lycorine derivatives and their preparation methods, pharmaceutical compositions containing them, and their use in the preparation of antiviral drugs.
- coronavirus treatment drugs such as Paxlovid and Molnupiravir
- the new coronavirus mutates too fast.
- Most of the drugs that have been launched or are about to be launched are nucleoside drugs with greater side effects and require a long period of treatment. It is easy to develop drug resistance in the future, and new structural types of therapeutic drugs are urgently needed.
- Lycorine was first isolated from the Amaryllidaceae plant Narcissus pseudonarcissus in 1877. It is widely distributed in the Amaryllidaceae plants. Due to its unique structure and rich pharmacological activities, lycorine is favored by medicinal chemists and pharmacologists. The pharmacological effects of lycorine include antiviral, anticancer, antiparasitic, antibacterial, anti-inflammatory, and inhibition of acetylcholinesterase. Among them, antiviral and anticancer activities have received particular attention.
- lycorine structural formula is as follows
- coronavirus SARS-CoV
- DEV dengue virus
- WNV West Nile virus
- retrovirus HIV-1
- HSV-1 herpes simplex virus
- PV poliovirus
- enterovirus EV71
- influenza virus H5N1
- HCV hepatitis C virus
- YFV yellow fever virus
- ZIKV Zika virus
- Recent studies have reported that lycorine has in vitro anti-coronavirus (SARS-CoV-2) activity, and its intensity of action is comparable to that of remdesivir.
- lycorine itself has pharmaceutical defects such as strong hydrophilicity, poor metabolic stability, and toxic and side effects
- lycorine broad-spectrum antiviral drugs is a top priority.
- the inventor also previously disclosed that a class of lycorine derivatives has anti-hand, foot and mouth virus activity (authorization announcement number CN 110759927 B, application date: July 27, 2018).
- the present invention focuses on the 1-position hydroxyl group of lycorine to carry out in-depth structural optimization exploration and obtains a type of lycorine derivatives.
- This type of compound shows good anti-coronavirus activity and has improved metabolic stability.
- the technical problem solved by the present invention is to provide a class of lycorine derivatives and their pharmaceutically acceptable salts and their preparation methods, as well as the use of their pharmaceutical compositions in the preparation of antiviral drugs.
- the present invention provides the following technical solutions:
- the first aspect of the technical solution of the present invention is to provide a class of lycorine derivatives represented by the following general formula (I) and pharmaceutically acceptable salts thereof:
- R 1 is hydrogen
- R 1 is hydrogen
- R 2 is C1-3 alkyl
- R 3 is C1-5 alkyl, phenoxy, phenylmercapto, substituted phenoxy, substituted phenylmercapto;
- substituents are selected from halogen, nitro, amino, acyl, cyano, methylmercapto, halomethyl, C1-3 alkyl, C1-3 alkoxy, C1-3 alkylamino;
- R 1 and R 2 are C1-3 alkyl
- R 3 is phenoxy or substituted phenoxy, phenylmercapto or substituted phenylmercapto;
- the substituent is selected from halogen, nitro, amino, acyl, cyano, methylmercapto, halomethyl, C1-3 alkyl, C1-3 alkoxy, C1-3 alkylamino;
- R 1 is hydrogen
- R 2 and R 3 are ring-forming compounds, and the ring-forming compounds are selected from C3-6 cycloalkanes, C3-6 heterocycloalkanes, substituted C 3-6 cycloalkanes, and C3-6 oxocycloalkanes;
- the substituent is selected from halogen, methyl, and hydroxyl.
- the heterocycloalkyl group contains at least one heteroatom selected from N, O, and S;
- the substituent of the substituted cycloalkane is selected from halogen and oxo;
- substitution position on the benzene ring is para-position, meta-position, or ortho-position, and the number of substituents is mono-substitution or poly-substitution.
- the halogen substituent is F, Cl, Br, I,.
- lycorine derivatives and pharmaceutically acceptable salts thereof are selected from the following compounds:
- Reaction step a first, lycorine is catalyzed by a deacidifying agent to generate 2-hydroxyTBS-protected lycorine; reaction step b: secondly, TBS-protected lycorine forms ester with 1-hydroxyl and organic acid under the action of a coupling agent; Reaction step c: Finally remove TBS protection to obtain the compound of general formula (I).
- reaction steps and reaction conditions are (a) TBSCl, imidazole, DMF, rt; (b) EDCI, DMAP, carboxylic acid, DCM; (c) 1) 12N HCl aq, EtOH, rt; 2) Na 2 CO 3 aq.
- the third aspect of the technical solution of the present invention is to provide a pharmaceutical composition, which comprises the lycorine derivative according to any one of claims 1 to 5 and a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.
- the compounds of the present invention may be administered orally, for example, in the form of capsules, tablets, powders, granules, syrups or similar dosage forms, or parenterally by injection, ointment, suppository or similar dosage form.
- These pharmaceutical preparations can be prepared by using adjuvants well known in the art, such as binders, excipients, stabilizers, disintegrants, flavoring agents, moisturizers, etc. Lubricants and the like are prepared by ordinary methods.
- the dosage will vary with the symptoms and age of the patient, the nature and severity of the disease or disorder and the route and mode of administration, in the case of oral administration to adult patients, the present invention
- the compound is normally administered at a total daily dose of 1 to 200 mg, preferably 5 to 50 mg, as a single dose, or in divided doses; for example, two or three times a day; in the case of intravenous injection, 0.1 may be administered in one to three divided doses per day. to 100 mg, preferably a dose of 5 to 50 mg.
- the fourth aspect of the technical solution of the present invention provides the lycorine derivatives described in the first aspect and pharmaceutically acceptable salts thereof or the viruses described in the application of the pharmaceutical composition described in the third aspect in the preparation of antiviral drugs. Selected from the new coronavirus SARS-CoV-2.
- the present invention focuses on providing lycorine derivatives represented by general formula (I), which mainly introduces different sterically hindered substituent groups on the 1-hydroxyl group of lycorine.
- lycorine derivatives represented by general formula (I) which mainly introduces different sterically hindered substituent groups on the 1-hydroxyl group of lycorine.
- anti-COVID-19 activity screening such derivatives have significant anti-COVID-19 activity, suggesting that such derivatives have good anti-viral prospects.
- DMEM Dulbecco's modified Eagle's medium
- FBS fetal bovine serum
- PE petroleum ether (60-90°C)
- SI Selection Index (CC 50 /EC 50 )
- the present invention discloses the application of a class of lycorine derivatives, including their salts, solvates, prodrugs and pharmaceutical compositions. Persons skilled in the art can learn from the contents of this article and appropriately improve the process parameters for implementation. It should be noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are deemed to be included in the present invention.
- the methods and applications of the present invention have been described through preferred embodiments. Relevant persons can obviously make modifications or appropriate changes and combinations to the methods and applications described herein without departing from the content, spirit and scope of the present invention to achieve and Apply the technology of this invention.
- Reagents and conditions (a) TBSCl, imidazole, DMF, rt; (b) EDCI, DMAP, carboxylic acid, DCM; (c) 1) 12N HCl aq, EtOH, rt; 2) Na 2 CO 3 aq.
- the crude intermediate II was dissolved in ethanol (5 mL), and then 4 mL of 12N concentrated hydrochloric acid was added and stirred at room temperature. After TLC monitoring of the reaction, add saturated Na 2 CO 3 aqueous solution to adjust the pH to greater than 9, extract with DCM (15 mL) three times, wash the combined organic phases three times with saturated NaCl aqueous solution (10 mL), dry over anhydrous Na 2 SO 4 , filter and concentrate. , the crude product is separated by column chromatography (eluent: PE-EA) to obtain the compound of general formula I.
- Vero cells at a density of 2 ⁇ 10 5 /mL were spread on a 96-well plate, 100 ⁇ L per well, and cultured overnight. Weigh appropriate amounts of lycorine and lycorine derivatives and prepare a 10mM stock solution with DMSO. For use, prepare the drug at 10 ⁇ M in DMEM or DMEM containing 2% FBS.
- CPE final cell pathology
- the SARS-CoV-2 N protein (Nucleocapsid protein) gene primer and FAM fluorescent probe were diluted to 5 ⁇ M, and the N plasmid was used as a standard (constructed by Peng Xiaozhong Laboratory), starting from 10 ng/ ⁇ L. A total of 8 concentration gradients were used to calculate the standard curve, and the copy number of the viral N gene was calculated based on the standard curve (absolute quantification). Continue to add the following components to the 384-well white bottom plate and centrifuge:
- Figure 1 shows that the initial screening results against the new coronavirus show that lycorine, LY152, LY176, LY198, LY199, LY202, LY203, LY205 and LY208 all have significant inhibitory effects on the new coronavirus strain GD108, with inhibition rates approaching or exceeding 100% at a dosage concentration of 10 ⁇ M, while compound LY150 has no inhibitory effect at 10 ⁇ M, suggesting that these effective derivatives can be further tested for their EC 50 values.
- Vero cells at a density of 2 ⁇ 10 5 /mL were spread on a 96-well plate, 100 ⁇ L per well, and cultured overnight. Weigh appropriate amounts of lycorine and lycorine derivatives and prepare a 10mM stock solution with DMSO. Use DMEM Or dilute the drug 2-fold (starting from 10 ⁇ M) in DMEM containing 2% FBS, for a total of 7 concentration gradients.
- the detection method is the same as Experimental Example 1. After calculating the inhibitory rate of different gradient concentrations of the compound on virus replication, use the statistical software GraphPad Prism 8 to draw a graph and calculate the half inhibitory concentration (EC 50 , 50% inhibition concentration) of the drug on viral N gene replication. ).
- Vero cells with a density of 2 ⁇ 10 5 /mL were added to a 96-well plate, 100 ⁇ L per well, and cultured overnight.
- Discard the culture medium in the 96-well plate add 200 ⁇ L DMEM diluted compound to each well, and incubate in a 37°C incubator for 48 hours.
- Discard the medium add 100 ⁇ L of DMEM medium containing CCK-8, and incubate at 37°C for 2 hours.
- the EC 50 values of the active compounds LY152 and LY176 against the new coronavirus mutant strains Alpha, Beta and Delta strains are all smaller than the positive drug remdesivir, and their EC 50 values against the Omicron strain are similar to remdesivir, indicating that LY152 , LY176 has application potential against new coronavirus mutant strains.
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Abstract
公开了一类如通式(I)所示的石蒜碱衍生物及其药学上可接受的盐,其制备方法、药物组合物及其应用。所述的石蒜碱衍生物具有明显的抗病毒活性,可用于治疗病毒性疾病。
Description
本发明属于医药技术领域,具体涉及一类石蒜碱衍生物及其制备方法,含有它们的药物组合物,和其在制备抗病毒药物中的用途。
自2019年12月以来新型冠状病毒疫情(COVID-19)已扩散至全球各个国家。该病毒主要经呼吸道飞沫和接触传播,对人群普遍易感并且可通过无症状感染者进行传播。据世界卫生组织(WHO)最新统计数据,截至目前,全球COVID-19确诊病例已超过5亿例,累计死亡超过600万例。多国面临具有超强传染能力的变异病毒产生的疫情多次暴发难题。病毒变异已经导致研发上市的新冠疫苗保护力下降或无效的情况发生。目前虽有帕罗维德(Paxlovid)、莫努匹韦(Molnupiravir)等新冠病毒治疗药物上市,但新冠病毒变异太快,已上市或即将上市的药物多为核苷类药物副作用较大且治疗一段时间后易产生耐药性,急需新结构类型治疗药物。
石蒜碱(Lycorine)于1877年首次从石蒜科植物洋水仙(Narcissus pseudonarcissus)中分离得到,其在石蒜科植物中有广泛的分布。因独特的结构和丰富的药理活性,石蒜碱备受药物化学家和药理学家青睐。石蒜碱的药理作用包含了抗病毒、抗癌、抗寄生虫、抗菌、抗炎、抑制乙酰胆碱酯酶等。其中,抗病毒和抗癌活性尤其受到广泛关注。
研究表明,石蒜碱(结构式如下)具有较强的广谱抗病毒作用,包括冠状病毒(SARS-CoV)、登革热病毒(DENV)、西尼罗河病毒(WNV)、逆转录病毒(HIV-1)、单纯疱疹病毒(HSV-1)、脊髓灰质炎病毒(PV)、肠道病毒(EV71)、流感病毒(H5N1)、丙肝病毒(HCV)、黄热病病毒(YFV)、寨卡病毒(ZIKV)等。近期研究报道,石蒜碱具有体外抗新冠病毒(SARS-CoV-2)活性,且作用强度与瑞德西韦相当。由于石蒜碱本身具有亲水性强、代谢稳定性差及毒副作用等成药性缺陷,因此,石蒜碱类广谱抗病毒药物的开发是当务之急。发明人前期亦公开了一类石蒜碱衍生物具有抗手足口病毒活性(授权公告号CN 110759927 B,申请日期:2018年7月27日)。
本发明聚焦石蒜碱的1-位羟基展开深入的结构优化探索,获得一类石蒜碱衍生物,该类化合物显示良好的抗新冠病毒活性,同时代谢稳定性有所改善。
参考文献:
[1]曾炳麟,等.石蒜碱药理活性及构效关系研究进展[J].天然产物研究与开发,2021,33(02):342-351.
[2]Wang HQ,et al.Lycorine derivative LY-55 inhibits EV71 and CVA16 replication through downregulating autophagy[J].Front Cell Infect Microbiol,2019,9:277-277.
发明内容
本发明解决的技术问题是提供一类石蒜碱衍生物及其药学上可接受的盐和其制备方法,及其药物组合物在制备抗病毒药物中的用途。
为解决本发明的技术问题,本发明提供如下技术方案:
本发明技术方案的第一方面是提供了一类具有下列通式(Ⅰ)所示的石蒜碱衍生物及其药学上可接受的盐:
其中:
R1为氢;
R1为氢;
R2为C1-3烷基;
R3为C1-5烷基,苯氧基,苯巯基,取代苯氧基,取代苯巯基;
上述取代基选自卤素、硝基、氨基、酰基、氰基、甲巯基、卤代甲基、C1-3烷基、C1-3烷氧基、C1-3烷氨基;
或
R1、R2为C1-3烷基;
R3为苯氧基或取代苯氧基,苯巯基或取代苯巯基;
所述取代基选自卤素、硝基、氨基、酰基、氰基、甲巯基、卤代甲基、C1-3烷基、C1-3烷氧基、C1-3烷氨基;
或
R1为氢,R2、R3成环化合物,所述成环化合物选自C3-6环烷烃,C3-6杂环烷烃,取代C 3-6环烷烃,C3-6氧代环烷烃;所述取代基选自卤素,甲基,羟基。
所述杂环烷基至少含有一个选自N、O、S中的杂原子;
所述取代环烷烃其取代基选自卤素,氧代;
所述苯环上的取代位置为对位、间位、邻位,取代基数目为单取代、多取代。
所述卤素取代基为F、Cl、Br、I,。
最优选的石蒜碱衍生物及其药学上可接受的盐,选自如下化合物:
本发明技术方案的第二方面是提供了上表所述的化合物的制备方法:
反应步骤a:首先石蒜碱在脱酸剂催化下生成2-羟基TBS保护的石蒜碱;反应步骤b:其次TBS保护的石蒜碱在偶合剂作用下1-羟基和有机酸成酯;反应步骤c:最后脱去TBS保护得到通式(Ⅰ)化合物。
最优选的反应步骤及反应条件为(a)TBSCl,咪唑,DMF,r.t.;(b)EDCI,DMAP,羧酸,DCM;(c)1)12N HCl aq,EtOH,r.t.;2)Na2CO3aq.
本发明技术方案的第三方面是提供了一种药物组合物,所述的药物组合物包含权利要求1-5任一项的石蒜碱衍生物及其药学上可接受的盐以及药学上可接受的载体或赋形剂。
本发明的化合物可以例如胶囊剂,片剂,粉剂,粒剂,糖浆或类似剂型形式口服给药,或通过注射,软膏,栓剂或类似剂型非肠胃给药。这些药物制剂可通过使用本领域熟知的辅助剂,如粘合剂,赋形剂,稳定剂,崩解剂,矫味剂,润
滑剂等等以普通方法生成,虽然剂量随症状和病人的年龄,疾病或失调的性质及严重性和给药的途径和方式而变,但对成年病人口服给药的情况来说,本发明化合物正常给药为每天总剂量1至200mg,优选为5至50mg,以为单剂量,或者为分剂量形式;例如每日二次或三次;对于静脉注射的情况,每天可以分一至三次给用0.1至100mg,优选为5至50mg的剂量。
本发明技术方案的第四方面是提供了第一方面所述石蒜碱衍生物及其药学上可接受的盐或第三方面所述药物组合物在制备抗病毒药物中的应用所述的病毒选自新冠病毒SARS-CoV-2。
有益技术效果
本发明重点提供具有通式(Ⅰ)所示的石蒜碱衍生物,主要在石蒜碱的1位羟基上引入了不同的位阻取代基团。通过抗新冠病毒活性筛选,此类衍生物具有显著的抗新冠病毒活性,提示此类衍生物具有较好的抗病毒前景。
图1石蒜碱衍生物对新冠病毒野生株GD108株初筛抑制情况(10μM)
图2衍生物对新冠病毒野生株GD108的EC50测定结果
图3石蒜碱衍生物半数毒性浓度(CC50)测定结果
图4衍生物对4种新冠变异病毒株的EC50测定结果
英文名及缩写词:
aq.:水溶液
CC50:半数毒性浓度
DCM:二氯甲烷
DMEM:达尔伯克(氏)改良伊格尔(氏)培养基
DMF:N,N-二甲基甲酰胺
DMSO:二甲基亚砜
DMAP:4-二甲胺基吡啶
EA:乙酸乙酯
EC50:50%抑制浓度
EDCI:1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐
EtOH:乙醇
FBS:胎牛血清
LY:石蒜碱
MOI:感染复数
OD:吸光度
PE:石油醚(60-90℃)
SI:选择指数(CC50/EC50)
TBSCl:叔丁基二甲基氯硅烷
Remdesivir:瑞德西韦
r.t.:室温
μM:μmol/L
本发明公开了一类石蒜碱衍生物,含有它们的盐、溶剂合物、前药与药物组合物的应用,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来是显而易见的,它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合,来实现和应用本发明技术。
下面结合实施例,进一步阐述本发明:
化合物通用合成方法:
反应式如下:
试剂和条件:(a)TBSCl,咪唑,DMF,r.t.;(b)EDCI,DMAP,羧酸,DCM;(c)1)12N HCl aq,EtOH,r.t.;2)Na2CO3aq.
将石蒜碱(287mg,1mmol)、咪唑(136mg,2mmol)、TBSCl(165mg,1.1mmol)溶于5mL无水DMF中,室温搅拌至TLC监测反应完全后,加饱和NaCl水溶液(20mL)稀释,DCM(10mL)萃取三次,合并有机相,再用饱和NaCl水溶液(15mL)洗三次,有机相经无水Na2SO4干燥,过滤,浓缩得淡黄色固体中间体I,
直接用于下一步投料。
将相应的有机酸(1.2mmol)、EDCI(383mg,2mmol)、DMAP(24mg,0.2mmol)溶于2mL无水DCM中,室温搅拌10min。向其中加入中间体I(约1mmol)溶解于3mLDCM的溶液,室温搅拌4h。反应结束后,反应液用DCM(30mL)稀释,用饱和NaCl水溶液(15mL)洗三次,有机相直接浓缩得中间体II直接用于下一步反应。
粗中间体II加入乙醇(5mL)溶解,再加入4mL的12N的浓盐酸室温搅拌。TLC监测反应结束后,加饱和Na2CO3水溶液调至pH大于9,DCM(15mL)萃取三次,合并有机相用饱和NaCl水溶液(10mL)洗三次,无水Na2SO4干燥,过滤,浓缩,粗产物经柱层析分离(洗脱剂:PE-EA)得通式I化合物。
实施例1 1-羟基石蒜碱的环己基甲酸酯(LY-152)
白色固体,总收率25%。1H NMR(400MHz,CDCl3)δ6.58(s,1H),6.54(s,1H),5.87(d,J=1.5Hz,2H),5.56(s,1H),5.49(s,1H),4.24-3.55(m,3H),3.48(d,J=13.9Hz,1H),3.31(dt,J=9.2,4.8Hz,1H),2.84(d,J=10.3Hz,1H),2.73(d,J=10.5Hz,1H),2.65-2.55(m,2H),2.38(t,J=8.9Hz,1H),2.13(tt,J=11.0,3.6Hz,1H),1.73-1.43(m,5H),1.36-1.01(m,5H);13C NMR(101MHz,CDCl3)δ175.77,146.46,146.15,143.34,129.23,127.29,117.65,107.29,105.07,100.95,72.26,69.47,61.76,56.88,53.79,42.88,39.34,28.81,28.77,28.68,25.69,25.22;ESI-MS:398[M+H]+。
实施例2 1-羟基石蒜碱的环戊基甲酸酯(LY-177)
白色固体,总收率31%。1H NMR(400MHz,CDCl3)δ6.62(s,1H),6.55(s,1H),5.88(s,2H),5.58(s,1H),5.53(s,1H),4.13(s,2H),3.49(s,1H),3.32(s,1H),3.10-2.23(m,
7H),1.76-1.37(m,8H);13C NMR(101MHz,CDCl3)δ176.52,146.53,146.24,144.33,126.78,117.05,107.36,105.13,101.03,72.49,69.77,61.82,57.01,53.87,43.88,39.63,30.05,29.86,28.79,25.64;ESI-MS:384[M+H]+。
实施例3 1-羟基石蒜碱的环丁基甲酸酯(LY-179)
白色固体,总收率33%。1H NMR(400MHz,CDCl3)δ6.59(s,1H),6.56(s,1H),5.90(q,J=1.5Hz,2H),5.63(s,1H),5.53(s,1H),4.19-4.13(m,2H),3.61-3.48(m,1H),3.38-3.29(m,1H),3.21–2.54(m,10H),2.50-2.30(brs,1H);13C NMR(101MHz,CDCl3)δ175.35,146.53,146.25,143.96,129.44,126.80,117.47,107.36,105.11,101.03,72.50,69.79,61.74,56.99,53.84,39.64,38.12,28.77,25.24,18.53;ESI-MS:370[M+H]+。
实施例4 1-羟基石蒜碱的环己基甲酸酯(LY-180)
白色固体,收率31%。1H NMR(400MHz,CDCl3)δ6.64(s,1H),6.56(s,1H),5.90(s,2H),5.60(s,1H),5.53(s,1H),4.20-4.08(m,2H),3.58-3.47(m,1H),3.40-3.29(m,1H),3.18-2.89(m,2H),2.87(d,J=10.1Hz,1H),2.76(s,1H),2.66-2.59(m,2H),2.53-2.29(brs,1H),2.14-2.05(m,2H),2.05-1.98(m,2H),1.90-1.72(m,2H);13C NMR(101MHz,CDCl3)δ173.00,146.59,146.40,143.81,129.44,126.74,118.70(dd,J=282,270Hz),117.35,107.45,104.81,101.09,73.44,69.43,61.74,56.95,53.81,39.45,38.68(t,J=24.4Hz),28.65,26.55(dd,J=13.8,6.2Hz),22.73;ESI-MS:406[M+H]+。
实施例5 1-羟基石蒜碱的四氢呋喃-2S-甲酸酯(LY-182)
白色固体,总收率40%。1H NMR(500MHz,CDCl3)δ6.68(s,1H),6.58(s,1H),6.00(s,2H),5.91(s,2H),4.17(d,J=13.8Hz,1H),3.59(d,J=10.8Hz,1H),3.47(s,1H),3.31(d,J=10.3Hz,1H),3.13(d,J=10.1Hz,1H),3.02-2.39(m,11H);13C NMR(101MHz,CDCl3)δ173.43,146.55,146.35,143.60,129.35,126.98,117.41,107.39,105.00,101.05,76.60,73.15,69.47,69.32,61.75,56.97,53.79,39.44,30.57,28.70,24.78,22.74;ESI-MS:386[M+H]+。
实施例6 1-羟基石蒜碱的2’S-氟代环丙基-(2R)-甲酸酯(LY-184)
白色固体,总收率38%。1H NMR(400MHz,CDCl3)δ6.65(s,1H),6.58(s,1H),5.91(s,2H),5.64(s,1H),5.54(s,1H),4.67-4.50(m,1H),4.17-4.12(m,2H),3.62-3.44(m,1H),3.35(s,1H),3.12-2.20(m,6H),1.83-1.57(m,2H),1.17-1.02(m,1H);13C NMR(101MHz,CDCl3)δ168.38,146.61,146.31,143.34,129.21,127.19,117.74,107.38,105.11,101.05,73.52,71.84(d,J=232.0Hz),69.49,61.64,56.80,53.79,39.16,28.66,20.03(d,J=11.1Hz),12.57(d,J=10.3Hz);ESI-MS:374[M+H]+。
实施例7 1-羟基石蒜碱的2-(4-氟苯巯基)丙酸酯(LY-176)
白色固体,总收率25%。1H NMR(400MHz,CDCl3)δ7.16-7.04(m,2H),6.86(t,J=8.5Hz,2H),6.65(s,1H),6.56(s,1H),5.90(d,J=3.3Hz,2H),5.62(s,1H),5.53(s,
1H),4.12(d,J=14.1Hz,2H),3.60-3.27(m,3H),2.85(s,1H),2.68-2.17(m,5H),1.28(d,J=7.2Hz,3H);13C NMR(101MHz,CDCl3)δ172.35,163.98,161.51,146.66,146.44,143.94,135.33,135.24,129.15,127.90,127.87,126.79,117.44,116.12,115.90,107.46,105.15,101.11,73.26,69.50,61.60,56.76,53.77,45.18,39.37,28.72,17.37;ESI-MS:470[M+H]+。
实施例8 1-羟基石蒜碱的2-甲基-2-(4-氯苯氧基)丙酸酯(LY-150)
白色固体,总收率30%。1H NMR(400MHz,CDCl3)δ6.93(d,J=8.9Hz,2H),6.61(s,1H),6.53(s,1H),6.45(d,J=8.9Hz,2H),5.95(d,J=20.6Hz,2H),5.65(s,1H),5.45(s,1H),4.12-4.07(m,2H),3.65-3.10(m,3H),2.82(d,J=10.4Hz,1H),2.54(s,2H),2.45(d,J=10.5Hz,1H),2.32-2.11(m,1H),1.42(s,3H),1.41(s,3H);13C NMR(101MHz,CDCl3)δ173.42,153.93,146.62,146.46,143.81,128.97,126.56,126.48,119.46,117.22,107.59,105.02,101.18,79.23,73.23,69.23,61.69,56.71,53.68,39.32,28.56,26.21,24.83;HRMS(ESI)m/z calcd.for C26H25O6NCl[M+H]482.13649,found482.13696.
实施例9 1-羟基石蒜碱的2-(4-氯苯氧基)丙酸酯(LY-198)
淡黄色固体,总收率22%。1HNMR(400MHz,CDCl3)δ7.11-6.86(m,2H),6.62-6.39(m,3H),5.97(d,J=16Hz,2H),5.66(s,1H),5.48(s,1H),4.57(s,1H),4.26-3.90(m,2H),3.59(s,1H),3.35(s,1H),2.98-2.14(m,6H),2.13-1.92(m,1H),1.47(s,3H);ESI-MS:470[M+H]+。
实施例10 1-羟基石蒜碱的异丁酸酯(LY-181)
白色固体,总收率45%。1H NMR(400MHz,CDCl3)δ6.62(s,1H),6.56(s,1H),5.88(s,2H),5.58(s,1H),5.53(s,1H),4.12(d,J=15.0Hz,2H),3.53(s,1H),3.32(s,1H),2.94-2.30(m,7H),1.00(d,J=7.0Hz,3H),0.94(d,J=7.0Hz,3H);13C NMR(101MHz,CDCl3)δ176.88,146.88,146.27,140.75,129.32,117.85,107.21,105.03,100.98,72.27,69.43,62.17,56.92,53.79,39.18,33.94,28.58,18.97,18.79;ESI-MS:358[M+H]+。
实施例11 1-羟基石蒜碱的3’-氧代环丁烷甲酸酯(LY-201)
白色固体,总收率33%。1HNMR(400MHz,CDCl3)δ6.60(s,1H),6.57(s,1H),5.90(s,2H),5.64(s,1H),5.54(s,1H),4.16(s,2H),3.58-3.45(m,1H),3.42-3.32(m,1H),3.04-2.29(m,11H);13C NMR(101MHz,CDCl3)δ203.70,173.25,146.58,143.41,129.32,126.32,117.54,107.34,105.43,101.01,98.65,73.84,72.78,69.36,61.68,57.60,51.99,39.22,36.22,28.62;ESI-MS:384[M+H]+。
药理实验
实验例1体外抗新冠病毒活性初筛(图1)
(1)化合物配制与攻毒实验
将密度为2×105/mL的Vero细胞铺于96孔板,每孔100μL,过夜贴壁培养。称取适量的石蒜碱和石蒜碱衍生物,用DMSO配制为10mM储存液。使用时用DMEM或含2%FBS的DMEM将药物配制为10μM。用DMEM稀释待测药物(浓度为10μM)处理Vero细胞1小时(96孔板,100μL/孔)后,加入病毒(100μL/孔)感染Vero细胞1小时(MOI=0.05),弃去药物-病毒混合物,重新加入2%FBS的DMEM稀释的药物(浓度为10μM),37℃培养箱孵育48小时。观察药物作用
后的细胞病变情况(CPE),取培养细胞上清液100μL,加200μL Trizol处理,用核酸提取试剂盒提取RNA进行后续实验。
(2)实时荧光定量检测病毒RNA拷贝数
将SARS-CoV-2的N蛋白(Nucleocapsid protein)基因引物和FAM荧光探针(由生工生物合成)稀释为5μM,N质粒作为标准品(彭小忠实验室构建),从10ng/μL开始进行十倍梯度稀释,共8个浓度梯度用于计算标准曲线,根据标准曲线计算出病毒N基因的拷贝数(绝对定量)。在384孔白底板中继续加入以下组分并离心:
获得总体积为10μL的反应体系,设置PCR仪按如下步骤运行:
仪器运行结束后会根据标准曲线计算出样品N基因拷贝数,通过公式计算出化合物对病毒的抑制率=(病毒对照组N基因拷贝数-药物组N基因拷贝数)/病毒组对照组N基因拷贝数×100%计算化合物的对病毒复制的抑制率。
图1抗新冠病毒初筛结果显示,石蒜碱、LY152、LY176、LY198、LY199、LY202、LY203、LY205及LY208都有对新冠病毒株GD108有显著抑制作用,在给药浓度10μM下抑制率接近或超过100%,而化合物LY150在10μM无抑制作用,提示这些有效衍生物可进一步测定其EC50值。
实验例2体外有效化合物EC50的测定(图2)
(1)化合物配制与攻毒实验
将密度为2×105/mL的Vero细胞铺于96孔板,每孔100μL,过夜贴壁培养。称取适量的石蒜碱和石蒜碱衍生物,用DMSO配制为10mM储存液。用DMEM
或含2%FBS的DMEM将药物进行2倍倍比稀释(从10μM开始稀释),共7个浓度梯度。用DMEM稀释待测药物处理Vero细胞1小时(96孔板,100μL/孔)后,加入病毒(100μL/孔)感染Vero细胞1小时(MOI=0.05),弃去药物-病毒混合物,重新加入2%FBS的DMEM稀释的药物,37℃培养箱孵育48小时。观察药物作用后的细胞病变情况(CPE),取培养细胞上清液100μL,加200μL Trizol处理,用核酸提取试剂盒提取RNA进行后续实验。
(2)实时荧光定量检测病毒RNA拷贝数
检测方法与实验例1相同,计算出化合物不同梯度浓度对病毒复制的抑制率后,用统计软件GraphPad Prism 8作图并计算药物对病毒N基因复制的半数抑制浓度(EC50,50%inhibition concentration)。
通过有效化合物EC50的测定,发现LY152和LY176对新冠病毒野生株GD108具有较小的EC50值(图2),和石蒜碱相近,并优于瑞德西韦(EC501.205μM)。
实验例3 EC50最有效化合物的CC50测定(CCK8法,图3)
化合物CC50测定方法如下:
将密度为2×105/mL的Vero细胞加入96孔板,每孔100μL,过夜贴壁培养。用DMEM(Dulbecco's modified Eagle's medium)或含2%FBS的DMEM培养基梯度稀释到50、25、12.5、6.25、3.13、1.56和0.78μM。弃去96孔板中的培养基,每孔加入200μL DMEM稀释的化合物,37℃培养箱中孵育48小时。弃去培养基,加入100μL含CCK-8的DMEM培养基,37℃孵育2小时。设置阴性对照孔(加入0.1%DMSO)和空白对照孔(无细胞,加入DMEM)。使用酶标仪在450nm波长下测量吸光度值,通过公式:抑制率=(阴性对照组-药物组)/(阴性对照组-空白组)×100%计算化合物的细胞抑制率,其中阴性为阴性对照组吸光度(OD)平均值,药物组为化合物组OD平均值,空白为空白组OD平均值,并用统计软件GraphPad prism 8计算CC50。
从图3可知,实验测得LY152和LY176对Vero细胞的半数毒性浓度(CC50)分别为79.81和71.07μM,显示其毒性优于石蒜碱(CC5053.31μM)。
实施例4活性化合物对新冠四种变异株(Alpha、Beta、Delta、Omicron)的抗新冠病毒活性(测定方法见实验例2,图4)
从图4可知,活性化合物LY152、LY176对新冠病毒变异株Alpha、Beta和Delta株其EC50值均小于阳性药瑞德西韦,对Omicron株其EC50值与瑞德西韦相近,预示LY152、LY176有抗新冠病毒变异株的应用潜力。
Claims (11)
- 一类具有下列通式(Ⅰ)所示的石蒜碱衍生物或其药学上可接受的盐:
其中:R1为氢;R2为C1-3烷基;R3为C1-5烷基,苯氧基,苯巯基,取代苯氧基,取代苯巯基;上述取代基选自卤素、硝基、氨基、酰基、氰基、甲巯基、卤代甲基、C1-3烷基、C1-3烷氧基、C1-3烷氨基;或R1、R2为C1-3烷基;R3为苯氧基或取代苯氧基,苯巯基或取代苯巯基;所述取代基选自卤素、硝基、氨基、酰基、氰基、甲巯基、卤代甲基、C1-3烷基、C1-3烷氧基、C1-3烷氨基;或R1为氢,R2、R3成环化合物,所述成环化合物选自C3-6环烷烃,C3-6杂环烷烃,取代C3-6环烷烃,C3-6氧代环烷烃;所述取代基选自卤素,甲基,羟基。 - 根据权利要求1的石蒜碱衍生物或其药学上可接受的盐,其特征在于,所述杂环烷基至少含有一个选自N、O、S中的杂原子。
- 根据权利要求2的石蒜碱衍生物或其药学上可接受的盐,其特征在于,取代苯氧基,取代苯巯基的取代基在苯基上的取代位置为对位、间位、邻位,取代基数目为单取代、双取代或多取代。
- 根据权利要求1的石蒜碱衍生物或其药学上可接受的盐,其特征在于,所述的卤素为F、Cl、Br、I。
- 根据权利要求1的石蒜碱衍生物或其药学上可接受的盐,其特征在于,所述的化合物选自如下:
- 制备权利要求1至5任一项所述化合物的方法为:
反应步骤a:首先石蒜碱在脱酸剂催化下生成2-羟基TBS保护的石蒜碱;反应步骤b:其次TBS保护的石蒜碱在偶合剂作用下1-羟基和有机酸成酯;反应步骤c:最后脱去TBS保护得到通式(Ⅰ)化合物。 - 一种药物组合物,其特征在于,所述的药物组合物包含权利要求1-5任一项的石蒜碱衍生物或其药学上可接受的盐以及药学上可接受的载体。
- 根据权利要求7的药物组合物,其特征在于,所述的药物组合物选自片剂、胶囊剂、丸剂、注射剂、滴眼剂、喷雾剂、软膏剂等。
- 根据权利要求7的药物组合物,其特征在于,所述的药物组合物选自控释给药剂型、缓释给药剂型、各种微粒给药系统。
- 如权利要求1-5任一项所述的石蒜碱衍生物或其药学上可接受的盐在制备抗病毒药物中的应用。
- 根据权利要求10的应用,其特征在于,所述的病毒选自新冠病毒SARS-CoV-2,冠状病毒SARS-CoV、手足口病毒EV71、柯萨奇病毒A16、寨卡病毒。
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CN113967211A (zh) * | 2021-12-15 | 2022-01-25 | 山东达因海洋生物制药股份有限公司 | 盐酸石蒜碱硫酯在制备抗冠状病毒药物中的应用 |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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CN110759927A (zh) * | 2018-07-27 | 2020-02-07 | 山东达因海洋生物制药股份有限公司 | 一类石蒜碱衍生物及其药物组合物和用途 |
CN113967211A (zh) * | 2021-12-15 | 2022-01-25 | 山东达因海洋生物制药股份有限公司 | 盐酸石蒜碱硫酯在制备抗冠状病毒药物中的应用 |
Non-Patent Citations (3)
Title |
---|
WANG HUIQIANG, GUO TINGTING, YANG YAJUN, YU LIAN, PAN XIANDAO, LI YUHUAN: "Lycorine Derivative LY-55 Inhibits EV71 and CVA16 Replication Through Downregulating Autophagy", FRONTIERS IN CELLULAR INFECTION MICROBIOLOGY, FRONTIERS RESEARCH FOUNDATION, CH, vol. 9, 7 August 2019 (2019-08-07), CH , XP093149964, ISSN: 2235-2988, DOI: 10.3389/fcimb.2019.00277 * |
YANG YA-JUN, LIU JIANG-NING, PAN XIAN-DAO: "Synthesis and antiviral activity of lycorine derivatives", JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH, GORDON AND BREACH, CHUR, CH, vol. 22, no. 12, 1 December 2020 (2020-12-01), CH , pages 1188 - 1196, XP093149962, ISSN: 1028-6020, DOI: 10.1080/10286020.2020.1844674 * |
ZENG, BINGLIN; ZHAO, RU; PAN, XIAN-DAO: "Research Progress on the Pharmacological Activities and Structure-activity Relationships of Lycorine", NATURAL PRODUCT RESEARCH AND DEVELOPMENT, GAI KAN BIANJIBU, CHENGDU, CN, vol. 33, no. 2, 31 December 2021 (2021-12-31), CN , pages 342 - 351, 341, XP009554074, ISSN: 1001-6880, DOI: 10.16333/j.1001-6880.2021.2.019 * |
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