CN112175008A - Tenofovir mixed phosphoramidate compound, pharmaceutical composition and application thereof - Google Patents
Tenofovir mixed phosphoramidate compound, pharmaceutical composition and application thereof Download PDFInfo
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- CN112175008A CN112175008A CN202011135023.5A CN202011135023A CN112175008A CN 112175008 A CN112175008 A CN 112175008A CN 202011135023 A CN202011135023 A CN 202011135023A CN 112175008 A CN112175008 A CN 112175008A
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- tenofovir
- mixed phosphoramidate
- inhibitor
- phosphoramidate compound
- compound
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- -1 phosphoramidate compound Chemical class 0.000 title claims abstract description 44
- VCMJCVGFSROFHV-WZGZYPNHSA-N tenofovir disoproxil fumarate Chemical compound OC(=O)\C=C\C(O)=O.N1=CN=C2N(C[C@@H](C)OCP(=O)(OCOC(=O)OC(C)C)OCOC(=O)OC(C)C)C=NC2=C1N VCMJCVGFSROFHV-WZGZYPNHSA-N 0.000 title claims abstract description 40
- 229960004556 tenofovir Drugs 0.000 title claims abstract description 34
- 239000008194 pharmaceutical composition Substances 0.000 title claims abstract description 11
- 239000003814 drug Substances 0.000 claims abstract description 22
- 150000001875 compounds Chemical class 0.000 claims abstract description 15
- 241000700721 Hepatitis B virus Species 0.000 claims abstract description 7
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 4
- 229910052739 hydrogen Inorganic materials 0.000 claims description 68
- 239000001257 hydrogen Substances 0.000 claims description 38
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 32
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 31
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 28
- 125000000217 alkyl group Chemical group 0.000 claims description 17
- 239000003112 inhibitor Substances 0.000 claims description 14
- 150000003839 salts Chemical class 0.000 claims description 14
- 125000004437 phosphorous atom Chemical group 0.000 claims description 13
- 229910052698 phosphorus Inorganic materials 0.000 claims description 13
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 9
- 241000700739 Hepadnaviridae Species 0.000 claims description 8
- 241000725303 Human immunodeficiency virus Species 0.000 claims description 8
- 208000036142 Viral infection Diseases 0.000 claims description 7
- 125000003118 aryl group Chemical group 0.000 claims description 7
- 150000005840 aryl radicals Chemical class 0.000 claims description 7
- 150000002431 hydrogen Chemical group 0.000 claims description 7
- 230000009385 viral infection Effects 0.000 claims description 7
- 241000712907 Retroviridae Species 0.000 claims description 5
- 102100034343 Integrase Human genes 0.000 claims description 4
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 claims description 4
- 229940124597 therapeutic agent Drugs 0.000 claims description 4
- 241000700605 Viruses Species 0.000 claims description 3
- 208000015181 infectious disease Diseases 0.000 claims description 3
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 3
- 230000003612 virological effect Effects 0.000 claims description 3
- 125000006176 2-ethylbutyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(C([H])([H])*)C([H])([H])C([H])([H])[H] 0.000 claims description 2
- 102100035875 C-C chemokine receptor type 5 Human genes 0.000 claims description 2
- 101710149870 C-C chemokine receptor type 5 Proteins 0.000 claims description 2
- 102100031650 C-X-C chemokine receptor type 4 Human genes 0.000 claims description 2
- 108010002459 HIV Integrase Proteins 0.000 claims description 2
- 108010010369 HIV Protease Proteins 0.000 claims description 2
- 229940099797 HIV integrase inhibitor Drugs 0.000 claims description 2
- 101000922348 Homo sapiens C-X-C chemokine receptor type 4 Proteins 0.000 claims description 2
- 210000000234 capsid Anatomy 0.000 claims description 2
- 230000003197 catalytic effect Effects 0.000 claims description 2
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 229940124784 gp41 inhibitor Drugs 0.000 claims description 2
- 239000003084 hiv integrase inhibitor Substances 0.000 claims description 2
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 claims description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 2
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 2
- 125000001971 neopentyl group Chemical group [H]C([*])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 2
- 239000002777 nucleoside Substances 0.000 claims description 2
- 150000003833 nucleoside derivatives Chemical class 0.000 claims description 2
- 239000002773 nucleotide Substances 0.000 claims description 2
- 125000003729 nucleotide group Chemical group 0.000 claims description 2
- 238000006116 polymerization reaction Methods 0.000 claims description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 230000000694 effects Effects 0.000 abstract description 20
- 208000002672 hepatitis B Diseases 0.000 abstract description 17
- 238000002360 preparation method Methods 0.000 abstract description 14
- 229940079593 drug Drugs 0.000 abstract description 11
- 238000000338 in vitro Methods 0.000 abstract description 4
- 230000029812 viral genome replication Effects 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 3
- 238000011161 development Methods 0.000 abstract description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 60
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical group N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 34
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 31
- LDEKQSIMHVQZJK-CAQYMETFSA-N tenofovir alafenamide Chemical compound O([P@@](=O)(CO[C@H](C)CN1C2=NC=NC(N)=C2N=C1)N[C@@H](C)C(=O)OC(C)C)C1=CC=CC=C1 LDEKQSIMHVQZJK-CAQYMETFSA-N 0.000 description 23
- 229960004946 tenofovir alafenamide Drugs 0.000 description 23
- 238000005160 1H NMR spectroscopy Methods 0.000 description 16
- 238000004679 31P NMR spectroscopy Methods 0.000 description 15
- 229930024421 Adenine Natural products 0.000 description 15
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 15
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical group C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 15
- 229960000643 adenine Drugs 0.000 description 15
- 125000006364 carbonyl oxy methylene group Chemical group [H]C([H])([*:2])OC([*:1])=O 0.000 description 15
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 15
- 125000005328 phosphinyl group Chemical group [PH2](=O)* 0.000 description 15
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 15
- 238000001228 spectrum Methods 0.000 description 15
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 9
- 229940024606 amino acid Drugs 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 208000030507 AIDS Diseases 0.000 description 5
- 229960000980 entecavir Drugs 0.000 description 5
- YXPVEXCTPGULBZ-WQYNNSOESA-N entecavir hydrate Chemical compound O.C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](CO)C1=C YXPVEXCTPGULBZ-WQYNNSOESA-N 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 206010059866 Drug resistance Diseases 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 230000000840 anti-viral effect Effects 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 102200094422 rs3812883 Human genes 0.000 description 3
- 102200129228 rs387907164 Human genes 0.000 description 3
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- 208000000674 Hepadnaviridae Infections Diseases 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 208000005074 Retroviridae Infections Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 231100000417 nephrotoxicity Toxicity 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- ATUGKXMUOXFMLK-UTONKHPSSA-N Cl.C(C(C)(C)C)OC([C@H](N)CC1=CC=CC=C1)=O Chemical compound Cl.C(C(C)(C)C)OC([C@H](N)CC1=CC=CC=C1)=O ATUGKXMUOXFMLK-UTONKHPSSA-N 0.000 description 1
- 208000004930 Fatty Liver Diseases 0.000 description 1
- 206010019708 Hepatic steatosis Diseases 0.000 description 1
- 208000005176 Hepatitis C Diseases 0.000 description 1
- 206010019842 Hepatomegaly Diseases 0.000 description 1
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 1
- 206010029155 Nephropathy toxic Diseases 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000007012 clinical effect Effects 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 230000000311 effect on hepatitis Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 208000010706 fatty liver disease Diseases 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 208000006443 lactic acidosis Diseases 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000007694 nephrotoxicity Effects 0.000 description 1
- 231100000380 osteotoxicity Toxicity 0.000 description 1
- 229960005190 phenylalanine Drugs 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 235000015497 potassium bicarbonate Nutrition 0.000 description 1
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 1
- 239000011736 potassium bicarbonate Substances 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 231100000240 steatosis hepatitis Toxicity 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- 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
-
- 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
- A61P31/18—Antivirals for RNA viruses for HIV
-
- 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
- A61P31/20—Antivirals for DNA viruses
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Communicable Diseases (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Veterinary Medicine (AREA)
- Oncology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- AIDS & HIV (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to the technical field of medicines, and discloses a tenofovir mixed phosphoramidate compound, a pharmaceutical composition thereof, a preparation method and application thereof, in particular to application of the compound for treating hepatitis B. Experiments prove that the compound has the activity of inhibiting HBV virus replication, and meanwhile, the tenofovir mixed phosphoramidate compound has the advantages of high in-vitro activity, large development coefficient and the like compared with the existing hepatitis B treatment medicament TAF (Virida), and can be used for developing the hepatitis B treatment medicament.
Description
Technical Field
The invention relates to a tenofovir mixed phosphoramidate compound and a salt thereof, in particular to a tenofovir mixed phosphoramidate compound and a salt thereof with HIV-1/HBV virus replication inhibition activity, a pharmaceutical composition thereof and an application thereof.
Background
The current medicines for treating hepatitis B, namely ETV (entecavir), TDF (Werrede) and TAF (Viridae), have advantages and disadvantages respectively since the market. Advantages of ETV: the side effect is small; the disadvantages are as follows: has certain drug resistance, and is forbidden for pregnant women; TDF has the advantages that: the antiviral effect is good, and the drug resistance is very low; the disadvantages are as follows: has certain renal toxicity, and has certain side effects on bones and kidneys after long-term application; the TAF has the advantages that: has the same antiviral effect as TDF, raised kidney and bone laboratory safety parameters, less medicine resistance and dosage as one twelfth of that of one generation of tenofovir. The virtide (TAF) is a novel oral administration scheme for treating hepatitis B, although the hepatitis B cannot be completely cured, the efficacy of the virtide is greatly improved, and the hepatitis B virus amount can be well controlled.
In clinical terms, TAF and TDF have the same clinical effects in treating hepatitis b, but smaller amounts of TAF, lower toxicity, and greater effect in drug resistance and combination therapy. However, because one molecule of phenol is dropped off after TAF enters a human body, TAF compound preparations gradually show non-negligible toxic and side effects since the market: lactic acidosis and severe hepatomegaly with fatty liver. Therefore, the development of analogues of TAF with independent intellectual property rights, maintaining the minor structural changes, maintaining the excellent antiviral activity of TAF, while having better stability in plasma and oral bioavailability is of great importance.
In conclusion, although the global AIDS/hepatitis B treatment drugs, TDF and TAF compound preparations of Gilidde company still occupy absolute advantages, the research on the TNF-based AIDS/hepatitis B treatment drugs still has great significance. Further improves the bioavailability of the human body, improves the activity, reduces the toxic and side effect, and has important value for fully playing the efficacy of treating hepatitis B and AIDS.
Disclosure of Invention
In the process of researching the prodrug of Tenofovir Alafenamide (TAF) analogue, the inventor finds that after the benzene ring of TAF is replaced by an amino NHR chain, the activity is improved, the selection coefficient SI of the biological activity is also improved, so that the dosage of a research sample can be further reduced compared with that of TAF, namely the toxicity of TAF can be further reduced, the activity of TAF is kept, and the drug concentration in tissue cells, particularly liver cells is obviously increased compared with that of TAF, so that the tenofovir mixed phosphoramidate compound can obviously improve the treatment effect on hepatitis B and AIDS and greatly reduce the nephrotoxicity and the bone toxicity caused by TDF or TAF. The following invention has been thus proposed.
The invention adopts the technical scheme that a tenofovir mixed phosphoramidate compound with a structure (Ia) and salts thereof are disclosed,
wherein: ra、Rb、RcAnd RdEach independently represents (C)6-C20) Aryl group, (C)1-C12) Alkyl or (C)6-C20) Aryl radical (C)1-C12) Any one of alkyl, ReAnd RfEach independently represents hydrogen or (C)1-C12) Any one of alkyl groups; and when R iscAnd RdSimultaneously being methyl or simultaneously being benzyl, ReAnd RfAt the same time is hydrogen, and RaIs equal to RbWhen the two amino acid esters bonded to the phosphorus atom are not in the S configuration at the same time.
The tenofovir mixed phosphoramidate compound or the pharmaceutically acceptable salt thereof is provided, wherein ReAnd RfAnd simultaneously hydrogen, the amino acid ester connected with the phosphorus atom is in an R configuration or an S configuration, and the structural formula of the tenofovir mixed phosphoramidate compound is selected from one of the following structural formulas:
wherein: ra、Rb、RcAnd RdEach independently represents (C)6-C20) Aryl group, (C)1-C12) Alkyl or (C)6-C20) Aryl radical (C)1-C12) Any one of alkyl groups; and when R iscAnd RdBoth being methyl or both being benzyl, and RaIs equal to RbWhen the two amino acid esters bonded to the phosphorus atom are not in the S configuration at the same time.
The tenofovir mixed phosphoramidate compound or the pharmaceutically acceptable salt thereof is provided, wherein the phosphorus atom is chiral phosphorus atom, preferably S(P)Configuration or R(P)One or two of the configurations, the structural formula of the tenofovir mixed phosphoramidate compound is selected from one of the following structural formulas:
wherein: ra、Rb、RcAnd RdEach independently represents (C)6-C20) Aryl group, (C)1-C12) Alkyl or (C)6-C20) Aryl radical (C)1-C12) Any one of alkyl groups; and when R iscAnd RdBoth being methyl or both being benzyl, and RaIs equal to RbWhen the two amino acid esters bonded to the phosphorus atom are not in the S configuration at the same time.
The tenofovir mixed phosphoramidate compound or the pharmaceutically acceptable salt thereof of the present invention is characterized in that R isaAnd RbEach independently represents any one of isopropyl, ethyl, isobutyl, neopentyl, n-butyl, cyclohexyl, methyl, tert-butyl or 2-ethylbutyl, and R iscAnd RdEach independently represents any one of methyl or benzyl, and R iseAnd RfAnd hydrogen, the structural formula of the tenofovir mixed phosphoramidate compound is selected from one of the following structural formulas:
the pharmaceutical composition comprises the tenofovir mixed phosphoramidate compound or the pharmaceutically acceptable salt thereof and an auxiliary material, wherein the auxiliary material is a pharmaceutically acceptable carrier or excipient.
The pharmaceutical composition of the invention, wherein the pharmaceutical composition further comprises a therapeutically effective amount of an additional therapeutic agent, wherein the additional therapeutic agent is at least one of a compound that inhibits HIV protease, a HIV non-nucleoside inhibitor of reverse transcriptase, a HIV nucleotide inhibitor of reverse transcriptase, a HIV integrase inhibitor, a gp41 inhibitor, a CXCR4 inhibitor, a gp120 inhibitor, a CCR5 inhibitor, a viral capsid polymerization inhibitor, or a non-catalytic site HIV integrase site inhibitor.
The invention discloses a preparation method of a tenofovir mixed phosphoramidate compound, which comprises the following steps:
under the condition of-20 to-80 ℃, tenofovir (C0P00) reacts with a phosphorylation reagent to generate C0P66, and then C0P66 reacts with amino acid ester hydrochloride HA3ace and amino acid ester hydrochloride HA3bdf to obtain the target compound (Ia).
The synthetic route of the preparation method is as follows:
wherein: ra、Rb、RcAnd RdEach independently represents (C)6-C20) Aryl group, (C)1-C12) Alkyl or (C)6-C20) Aryl radical (C)1-C12) Any one of alkyl, ReAnd RfEach independently represents hydrogen or (C)1-C12) Any one of alkyl groups; and when R iscAnd RdSimultaneously being methyl or simultaneously being benzyl, ReAnd RfAt the same time is hydrogen, and RaIs equal to RbWhen the two amino acid esters bonded to the phosphorus atom are not in the S configuration at the same time.
The tenofovir mixed phosphoramidate compound or the pharmaceutically acceptable salt thereof is applied to the preparation of the medicines for treating human hepadnaviridae or retrovirus infection.
The use of the invention, wherein the human hepadnaviridae or retroviridae viral infection is a human HBV viral infection or HIV viral infection.
The invention also discloses application of the pharmaceutical composition in preparing a medicament for resisting human hepadnaviridae or retrovirus infection, wherein the human hepadnaviridae or retrovirus virus is HBV virus or HIV virus.
The invention has the beneficial effects that:
the tenofovir mixed phosphoramidate compound of the invention has excellent properties required by being a medicament for treating hepatitis C, which is determined by a detection mechanism, and comprises the following specific steps:
in the in-vitro anti-hepatitis B virus active screening, IC combined with C371S374S-2, C374R374R and C371R371R50Is 2-5 times of TAF (positive control), and the selection coefficient of biological activity SI is 2-5 times of TAF (positive control).
This indicates that: compared with a class of anti-hepatitis B drugs TAF, the tenofovir mixed phosphoramidate compound has high activity of inhibiting virus replication, has large selection coefficient of developing biological activity, and is expected to become a drug for treating HBV infection.
In a word, the tenofovir mixed phosphoramidate compound of the invention integrates various good attributes such as high activity, low toxicity, high bioavailability and the like, and has a prospect of becoming a new generation of drugs for treating AIDS or hepatitis B.
The specific implementation mode is as follows:
the following examples are presented to enable one of ordinary skill in the art to more fully understand the present invention and are not intended to limit the invention in any way. All compounds have the structure1H NMR or MS determination.
The raw materials used in the invention are as follows: tenofovir, L-phenylalanine ester hydrochloride, fumaric acid were all commercially available.
Example 1
Preparation of 9- [ (R) -2- [ [ (R) - [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (R) - (1-neopentyloxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C374S373R-1) and 9- [ (R) -2- [ [ (S) - [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (R) - (1-neopentyloxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C373S 373R-2):
a) preparation of 9- [ (R) -2- [ [ [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (R) - (1-neopentyloxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C374S373R)
Weighing 7.2g (25mmol) of commercially available tenofovir (C0P00), adding into a 250ml flask, adding 50g of thionyl chloride, heating to 55 ℃ for 10 minutes, then continuing to heat to 70 ℃ and stirring for 1 hour, distilling thionyl chloride under reduced pressure, adding 100ml of dry acetonitrile after distillation is finished, refluxing at 85 ℃ for 10 minutes, evaporating acetonitrile under reduced pressure, cooling to room temperature, then adding 50ml of dry dichloromethane and 50ml of dry acetonitrile, continuing to stir for 0.5 hour, then placing at-80 ℃, adding 6.44g (25mmol, 1eq) of L-n-butyl phenylalanine hydrochloride (L-374 AH) after 0.5 hour, after the addition is finished, stirring for 10 minutes, dropping about 20ml of triethylamine, finishing the dropping, and continuing to stir for 0.5 hour; 6.79g (25mmol, 1eq) of D-phenylalanine neopentyl ester hydrochloride (D-AH373) were then added and stirred for a further 10min, approximately 20ml of triethylamine were added dropwise and stirring was continued for 0.5 h.
Suction filtration is carried out, and 100ml of ethyl acetate is added after the filtrate is evaporated to dryness. And washed twice with 100g of 5% potassium bicarbonate aqueous solution and twice with 100g of 20% sodium chloride aqueous solution respectively, dried by anhydrous sodium sulfate, filtered by suction, and the filtrate is evaporated to dryness. Ethyl acetate column chromatography gave about 7g (98.2% purity) of C374S 373R.
Nuclear magnetic hydrogen spectrum data of C374S 373R:1H NMR(400MHz，CDCl3)(ppm)： 0.80-1.15(15H，m，5×CH3)，1.21-1.43(2H，m，CH2)，1.57-1.77(2H， m，CH2)，2.76-2.95(1H,m,OCH),3.18-3.40(2H,m,OCH2P)， 3.82-4.00(8H，m，2×CH2，2×NH and 2×NCH)，4.13-4.38(6H，m， NCH2，2×COOCH2)， 6.29(2H,s,NH2) 7.90(1H, s, H on the purine ring), 7.06-7.39(10H, m, hydrogen on both benzene rings), 8.34(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：4.2，6.6；
LCMS-ESI+(m/z):708.8(M+H)。
b) The diastereomer mixture (C374S373R) was resolved by batch elution chromatography using a preparative column under the following elution conditions to give optically pure isomer C374S373R-1 and optically pure isomer C374S 373R-2.
Nuclear magnetic hydrogen spectrum data of C374S 373R-1:1H NMR(400MHz，CDCl3)(ppm)： 0.78-1.12(15H，m，5×CH3)，1.20-1.40(2H，m，CH2)，1.57-1.75(2H， m，CH2)，2.75-2.90(1H,m,OCH),3.18-3.30(2H,m,OCH2P)， 3.80-4.00(8H，m，2×CH2，2×NH and 2×NCH)，4.10-4.35(6H，m， NCH2，2×COOCH2)，6.27(2H,s,NH2) 7.89(1H, s, H on the purine ring), 7.04-7.37(10H, m, hydrogen on both benzene rings), 8.28(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：4.2；
LCMS-ESI+(m/z):708.8(M+H)。
Nuclear magnetic hydrogen spectrum data of C374S 373R-2:1H NMR(400MHz，CDCl3)(ppm)： 0.85-1.18(15H，m，5×CH3)，1.24-1.46(2H，m，CH2)，1.59-1.79(2H， m，CH2)，2.78-2.98(1H,m,OCH),3.28-3.44(2H,m,OCH2P)， 3.85-4.04(8H，m，2×CH2，2×NH and 2×NCH)，4.16-4.40(6H，m， NCH2，2×COOCH2)， 6.33(2H,s,NH2) 7.93(1H, s, H on the purine ring), 7.09-7.39(10H, m, hydrogen on both benzene rings), 8.36(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：6.6；
LCMS-ESI+(m/z):708.8(M+H)。
Example 2
Preparation of 9- [ (R) -2- [ [ (R) - [ [ (R) - (1-neopentyloxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (R) - (1-tert-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C373R378R-1) and 9- [ (R) -2- [ [ (S) - [ [ (R) - (1-neopentyloxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (R) - (1-tert-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C373R 378R-2):
C373R378R-1 and C373R378R-2 were prepared in a similar manner to example 1.
Nuclear magnetic hydrogen spectrum data of C373R 378R-1:1H NMR(400MHz，CDCl3) (ppm)：0.80-1.26(21H，m，7×CH3)，2.74-2.98(1H,m,OCH), 3.28-3.47(2H,m,OCH2P)，3.70-3.98(8H，m，2×CH2，2×NH and 2×NCH)， 4.08-4.37(4H，m，NCH2 and COOCH2)，6.26(2H,s,NH2) 7.07-7.39(10H, m, hydrogen on both phenyl rings), 7.82(1H, s, H on the purine ring), 8.33(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：4.6；
LCMS-ESI+(m/z):708.8(M+H)。
Nuclear magnetic hydrogen spectrum data of C373R 378R-2:1H NMR(400MHz，CDCl3) (ppm)：0.81-1.28(21H，m，7×CH3)，2.75-2.99(1H,m,OCH), 3.31-3.47(2H,m,OCH2P)，3.73-3.99(8H，m，2×CH2，2×NH and 2×NCH)， 4.11-4.38(4H，m，NCH2 and COOCH2)，6.28(2H,s,NH2) 7.10-7.39(10H, m, hydrogen on both phenyl rings), 7.82(1H, s, H on the purine ring), 8.33(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：5.8；
LCMS-ESI+(m/z):708.8(M+H)。
Example 3
Preparation of 9- [ (R) -2- [ [ (R) - [ [ (R) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C374R374S-1) and 9- [ (R) -2- [ [ (S) - [ [ (R) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (374C 374R 374S-2):
C374R374S-1 and C374R374S-2 were prepared in a similar manner to example 1.
Nuclear magnetic hydrogen spectrum data of C374R 374S-1:1H NMR(400MHz，CDCl3)(ppm)： 0.81-1.15(9H，m，3×CH3)，1.19-1.44(4H，m，2×CH2)，1.55-1.70(4H， m，2×CH2)，2.76-2.97(1H,m,OCH),3.21-3.50(2H,m,OCH2P)， 3.90-4.04(8H，m，2×CH2，2×NH and 2×NCH)，4.13-4.37(6H，m，NCH2， 2×COOCH2)，6.28(2H,s,NH2) 7.05-7.37(10H, m, hydrogen on both phenyl rings) 7.86(1H, s, H on the purine ring), 8.31(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：3.2；
LCMS-ESI+(m/z):694.8(M+H)。
Nuclear magnetic hydrogen spectrum data of C374R 374S-2:1H NMR(400MHz，CDCl3)(ppm)： 0.82-1.17(9H，m，3×CH3)，1.21-1.47(4H，m，2×CH2)，1.57-1.74(4H， m，2×CH2)，2.78-2.99(1H,m,OCH),3.33-3.50(2H,m,OCH2P)，3.92-4.07(8H，m，2×CH2，2×NH and 2×NCH)，4.15-4.39(6H，m，NCH2， 2×COOCH2)，6.28(2H,s,NH2) 7.05-7.39(10H, m, hydrogen on both phenyl rings) 7.86(1H, s, H on the purine ring), 8.31(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：4.8；
LCMS-ESI+(m/z):694.8(M+H)。
Example 4
Preparation of 9- [ (R) -2- [ [ (R) - [ [ (S) - (1-ethoxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C371S374S-1) and 9- [ (R) -2- [ [ (S) - [ [ (S) - (1-ethoxycarbonyl-2-phenyl) ethyl ] amino ] [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C371S 374S-2):
C371S374S-1 and C371S374S-2 were prepared in a similar manner to example 1.
Nuclear magnetic hydrogen spectrum data of C371S 374S-1:1H NMR(400MHz，CDCl3)(ppm)： 1.02-1.13(3H,d,CH3)，1.17-1.41(8H，m，2×CH3 and CH2)， 1.59-1.71(2H，m，CH2)，2.77-2.98(1H,m,OCH),3.23-3.41(2H,m, OCH2P)，3.86-4.05(8H，m，2×CH2，2×NH and 2×NCH)，4.13-4.37(6H， m，NCH2 and 2×COOCH2)，6.28(2H,s,NH2) 7.04-7.35(10H, m, hydrogen on both phenyl rings), 7.86(1H, s, H on the purine ring), 8.36(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：3.8；
LCMS-ESI+(m/z):666.7(M+H)。
Nuclear magnetic hydrogen spectrum data of C371S 374S-2:1H NMR(400MHz，CDCl3)(ppm)：1.03-1.15(3H,d,CH3)，1.19-1.44(8H，m，2×CH3 and CH2)， 1.61-1.75(2H，m，CH2)，2.79-2.98(1H,m,OCH),3.20-3.40(2H,m, OCH2P)，3.85-4.07(8H，m，2×CH2，2×NH and 2×NCH)，4.16-4.39(6H， m，NCH2 and 2×COOCH2)，6.28(2H,s,NH2) 7.09-7.35(10H, m, hydrogen on both phenyl rings), 7.86(1H, s, H on the purine ring), 8.36(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：5.1；
LCMS-ESI+(m/z):666.7(M+H)。
Example 5
Preparation of 9- [ (R) -2- [ [ (R) - [ [ (R) - (1- (2-ethylbutyloxycarbonyl) -2-phenyl) ethyl ] amino ] [ [ (S) - (1- (2-ethylbutyloxycarbonyl) -2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C379R379S-1) and 9- [ (R) -2- [ [ (S) - [ [ (R) - (1- (2-ethylbutyloxycarbonyl) -2-phenyl) ethyl ] amino ] [ [ (S) - (1- (2-ethylbutyloxycarbonyl) -2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C379R37 379S-2):
C379R379S-1 and C379R379S-2 were prepared in a similar manner to example 1.
Nuclear magnetic hydrogen spectrum data of C379R 379S-1:1H NMR(400MHz，CDCl3) (ppm)：0.82-1.16(15H，m，5×CH3)，1.20-1.37(8H，m，4×CH2)，1.93-2.12 (2H，m，2×CH)，2.77-2.95(1H,m,OCH),3.31-3.50(2H,m,OCH2P)， 3.74-3.99(8H，m，2×CH2，2×NH and 2×NCH)，4.09-4.35(6H，m， 2×COOCH2 and NCH2)，6.31(2H,s,NH2) 7.11-7.38(10H, m, hydrogen on both phenyl rings), 7.82(1H, s, H on the purine ring), 8.35(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：4.8；
LCMS-ESI+(m/z):750.9(M+H)。
Nuclear magnetic hydrogen spectrum data of C379R 379S-2:1H NMR(400MHz，CDCl3) (ppm)：0.85-1.14(15H，m，5×CH3)，1.22-1.37(8H，m，4×CH2)， 1.93-2.15(2H，m，2×CH)，2.78-2.97(1H,m,OCH),3.33-3.50(2H,m, OCH2P)，3.77-3.99(8H，m，2×CH2，2×NH and 2×NCH)，4.09-4.35(6H， m，2×COOCH2 and NCH2)，6.31(2H,s,NH2) 7.10-7.38(10H, m, hydrogen on both phenyl rings), 7.82(1H, s, H on the purine ring), 8.35(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：5.2；
LCMS-ESI+(m/z):750.9(M+H)。
Example 6
Preparation of 9- [ (R) -2- [ [ (R) - [ [ (R) - (1-isobutoxycarbonyl) ethyl ] amino ] [ [ (S) - (1-isobutoxycarbonyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C32R32S-1) and 9- [ (R) -2- [ [ (S) - [ [ (R) - (1-isobutoxycarbonyl) ethyl ] amino ] [ [ (S) - (1-isobutoxycarbonyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C32R 32S-2):
C32R32S-1 and C32R32S-2 were prepared in a similar manner to example 1.
Nuclear magnetic hydrogen spectrum data of C32R 32S-1:1H NMR(400MHz，CDCl3)(ppm)： 0.90-1.31(21H，m，7×CH3)，2.38-2.50(2H，m，2×CH)， 2.77-2.93(1H,m,OCH),3.22-3.35(2H,m,OCH2P)，3.76-3.90(4H，m， 2×NH and 2×NCH)，4.08-4.35(6H，m，NCH2 and 2×COOCH2)， 6.27(2H,s,NH2) 7.87(1H, s, H on the purine nucleus), 8.36(1H, s, H on the purine nucleus).
31P NMR(162MHz,CDCl3)：3.5；
LCMS-ESI+(m/z):542.6(M+H)。
Nuclear magnetic hydrogen spectrum data of C32R 32S-2:1H NMR(400MHz，CDCl3)(ppm)： 0.92-1.33(21H，m，7×CH3)，2.38-2.55(2H，m，2×CH)， 2.75-2.94(1H,m,OCH),3.22-3.35(2H,m,OCH2P)，3.77-3.92(4H，m， 2×NH and 2×NCH)，4.06-4.36(6H，m，NCH2 and 2×COOCH2)， 6.27(2H,s,NH2) 7.87(1H, s, H on the purine nucleus), 8.36(1H, s, H on the purine nucleus).
31P NMR(162MHz,CDCl3)：5.0；
LCMS-ESI+(m/z):542.6(M+H)。
Example 7
Preparation of 9- [ (R) -2- [ [ (R) - [ [ (S) - (1-isobutoxycarbonyl) ethyl ] amino ] [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C32S374S-1) and 9- [ (R) -2- [ [ (S) - [ [ (S) - (1-isobutoxycarbonyl) ethyl ] amino ] [ [ (S) - (1-n-butoxycarbonyl-2-phenyl) ethyl ] amino ] phosphinyl ] methoxy ] propyl ] adenine (C32S 374S-2):
C32S374S-1 and C32S374S-2 were prepared in a similar manner to example 1.
Nuclear magnetic hydrogen spectrum data of C32S 374S-1:1H NMR(400MHz，CDCl3)(ppm)： 0.90-1.33(15H，m，5×CH3)，1.20-1.41(2H，m，CH2)，1.58-1.70(2H， m，CH2)，2.44-2.51(1H，m，CH)，2.77-2.98(1H,m,OCH),3.34-3.51(2H, m,OCH2P)，3.80-4.06(6H，m，CH2，2×NH and 2×NCH)，4.19-4.33(6H， m，NCH2，2×COOCH2)，6.24(2H,s,NH2) 7.05-7.35(5H, m, hydrogen on the benzene ring), 7.85(1H, s, H on the purine ring), 8.35(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：4.1；
LCMS-ESI+(m/z):618.7(M+H)。
Nuclear magnetic hydrogen spectrum data of C32S 374S-2:1H NMR(400MHz，CDCl3)(ppm)： 0.90-1.35(15H，m，5×CH3)，1.20-1.41(2H，m，CH2)，1.58-1.72(2H， m，CH2)，2.47-2.52(1H，m，CH)，2.78-2.98(1H,m,OCH),3.39-3.50(2H, m,OCH2P)，3.84-4.08(6H，m，CH2，2×NH and 2×NCH)，4.19-4.38(6H， m，NCH2，2×COOCH2)，6.24(2H,s,NH2) 7.07-7.35(5H, m, hydrogen on the benzene ring), 7.85(1H, s, H on the purine ring), 8.35(1H, s, H on the purine ring).
31P NMR(162MHz,CDCl3)：5.4；
LCMS-ESI+(m/z):618.7(M+H)。
Example 8
In vitro assay for anti-HBV viral Activity of Compounds of the invention
1. In vitro cell model HepG2.2.15 cells
Determination of anti-hepatitis B virus activity of compound by Dot blot method
2.1 HepG2.2.15 cells (4X 10)4Cells/well) to 96-well plates in
Incubated at 37 ℃ 5% overnight.
2.2 day after, compounds were diluted and different concentrations of compounds were added to the culture wells. The final concentration of DMSO in the culture was 1%. 1 μ M Entecavir (ETV) as 100%
Inhibition; DMSO at 1% served as 0% Inhibition control.
2.3 day five, the fresh medium containing the compound was replaced.
And 2.4, on the eighth day and the ninth day, removing the culture solution in the culture wells, and collecting the cells for dot hybridization.
3. As a result: see the following table-in vitro anti-hepatitis B virus activity screening table
Table one: cytotoxicity of Compounds, extracellular anti-HBV Activity (HepG2.2.15)
| Comp. | CC50(μM) | Anti-HBV replication IC50(nM) |
| C371S374S-2 | >10 | ++ |
| C379R379S-1 | >10 | + |
| C374R374R | >10 | ++ |
| C371R371R | >10 | ++ |
| TAF (Positive control) | >10 | + |
, + ++ refers to 0.1-1 nM; + represents 1-5 nM; + represents 5-10 nM;
4. and (4) experimental conclusion:
4.1 positive compound TAF accords with the expected inhibition effect on the replication of hepatitis B virus in HepG2.2.15 cells, and proves that the experiment is effective and credible.
4.2 anti-hepatitis B virus activity. In dot blot experiments, the activity of 3 test compounds in the above table against hepatitis B virus was 2-5 times higher than that of TAF (positive control).
4.3 this fully indicates: compared with the existing anti-hepatitis B drug TAF, the compound of the invention has higher activity of inhibiting virus replication, and is expected to become a new generation drug for treating HBV infection.
While the present invention has been described in considerable detail and with particular reference to a few illustrative embodiments thereof, it is not intended to be limited to any such details or embodiments or any particular embodiments, but it is to be construed as effectively covering the intended scope of the disclosure by providing broad, potential interpretations of such claims in view of the prior art with reference to the appended claims. Furthermore, the foregoing describes the disclosure in terms of embodiments foreseen by the inventor for which an enabling description was available, notwithstanding that insubstantial modifications of the disclosure, not presently foreseen, may nonetheless represent equivalent modifications thereto.
Claims (9)
1. A tenofovir mixed phosphoramidate compound or a pharmaceutically acceptable salt thereof, wherein the structure of the tenofovir mixed phosphoramidate compound is (Ia):
wherein: ra、Rb、RcAnd RdEach independently represents (C)6-C20) Aryl group, (C)1-C12) Alkyl or (C)6-C20) Aryl radical (C)1-C12) Any one of alkyl, ReAnd RfEach independently represents hydrogen or (C)1-C12) Any one of alkyl groups; and when R iscAnd RdSimultaneously being methyl or simultaneously being benzyl, ReAnd RfAt the same time is hydrogen, and RaIs equal to RbWhen the two amino acid esters bonded to the phosphorus atom are not in the S configuration at the same time.
2. The tenofovir mixed phosphoramidate compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R iseAnd RfAnd simultaneously hydrogen, the amino acid ester connected with the phosphorus atom is in an R configuration or an S configuration, and the structural formula of the tenofovir mixed phosphoramidate compound is selected from one of the following structural formulas:
wherein: ra、Rb、RcAnd RdEach independently represents (C)6-C20) Aryl group, (C)1-C12) Alkyl or (C)6-C20) Aryl radical (C)1-C12) Any one of alkyl groups; and when R iscAnd RdBoth being methyl or both being benzyl, and RaIs equal to RbWhen the two amino acid esters bonded to the phosphorus atom are not in the S configuration at the same time.
3. Tenofovir mixed phosphoramidate compound or pharmaceutically acceptable salt thereof according to claim 2 wherein the phosphorus atom is a chiral phosphorus atom, preferably S(P)Configuration or R(P)One or two of the configurations, the structural formula of the tenofovir mixed phosphoramidate compound is selected from one of the following structural formulas:
wherein: ra、Rb、RcAnd RdEach independently represents (C)6-C20) Aryl group, (C)1-C12) Alkyl or (C)6-C20) Aryl radical (C)1-C12) Any one of alkyl groups; and when R iscAnd RdBoth being methyl or both being benzyl, and RaIs equal to RbWhen the two amino acid esters bonded to the phosphorus atom are not in the S configuration at the same time.
4. The tenofovir mixed phosphoramidate compound or pharmaceutically acceptable salt thereof according to any of claims 1-3, wherein R isaAnd RbEach independently represents any one of isopropyl, ethyl, isobutyl, neopentyl, n-butyl, cyclohexyl, methyl, tert-butyl or 2-ethylbutyl, and R iscAnd RdEach independently represents any one of methyl or benzyl, and R iseAnd RfAnd hydrogen, the structural formula of the tenofovir mixed phosphoramidate compound is selected from one of the following structural formulas:
5. a pharmaceutical composition comprising the tenofovir mixed phosphoramidate compound or pharmaceutically acceptable salt thereof according to any of claims 1-4 and an excipient which is a pharmaceutically acceptable carrier or excipient.
6. The pharmaceutical composition of claim 5, further comprising a therapeutically effective amount of an additional therapeutic agent, wherein the additional therapeutic agent is at least one of an HIV protease inhibiting compound, an HIV non-nucleoside inhibitor of reverse transcriptase, an HIV nucleotide inhibitor of reverse transcriptase, an HIV integrase inhibitor, a gp41 inhibitor, a CXCR4 inhibitor, a gp120 inhibitor, a CCR5 inhibitor, a viral capsid polymerization inhibitor, or a non-catalytic site HIV integrase site inhibitor.
7. Use of a tenofovir mixed phosphoramidate compound or a pharmaceutically acceptable salt thereof according to any of claims 1-4 for the manufacture of a medicament for the treatment of a hepadnaviridae or retroviridae viral infection in a human.
8. The use according to claim 7, wherein the human hepadnaviridae or retroviridae viral infection is a human HBV viral infection or HIV viral infection.
9. Use of a pharmaceutical composition according to any one of claims 5 to 6 for the manufacture of a medicament against infection by a virus of the human hepadnaviridae or retroviridae family, wherein the virus of the human hepadnaviridae or retroviridae family is an HBV virus or an HIV virus.
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