CN108640959A - Bis- dehydrogenation nucleoside compound of 3`- deoxidations -3`, 4`- and its application - Google Patents
Bis- dehydrogenation nucleoside compound of 3`- deoxidations -3`, 4`- and its application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
- C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
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- 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
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- 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
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- 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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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
- C07H19/207—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids the phosphoric or polyphosphoric acids being esterified by a further hydroxylic compound, e.g. flavine adenine dinucleotide or nicotinamide-adenine dinucleotide
Abstract
The present invention relates to antiviral fields, and in particular to the preparation of nucleoside compound (I) and its application in anti HIV-1 virus, hepatitis (HCV) virus, hepatitis B (HBV) virus, hydrophobin and zika virus.
Description
Technical field
Viral infection is being treated or prevented the present invention relates to bis- dehydrogenation nucleoside compound of 3'- deoxidations -3', 4'- is used
The application of aspect.It is being treated or prevented more specifically, the present invention describes bis- dehydrogenation nucleoside compound of 3'- deoxidations -3', 4'-
By rabies viruses, inhibition of HIV, HCV virus, HBV viruses, the purposes of zika virus infection.
Background technology
Nucleoside compound has more than 10 kinds and is approved for treatment human immunodeficiency virus (HIV) or the third type at present
Hepatitis virus (HCV) and other viruses.Challenge in developing antiviral therapy is that suppressing virus replication is thin without damaging host
Born of the same parents.In general, in order to show antiviral activity, nucleoside analog must have host cell enzymes metabolic conversion be they corresponding three
Phosphoric acid.The triphosphate forms, nucleoside polymerase inhibitor simulates natural nucleotide, because they are naturally occurring with five kinds
One kind (CTP, UTP, TTP, ATP or GTP) competition of nucleoside 5 '-triphosphoric acid (NTP) extend for RNA or DNA.Therefore core
Glycosides analog is by being used as the chain terminator of chain terminator or delay come suppressing virus replication.
Acquired immunodeficiency syndrome (AIDS) is to infect a kind of caused serious harm people's life by inhibition of HIV to be good for
The communicable disease of health.Global AIDS patient is counted more than 40,000,000 according to WHO, increases patient 5,000,000 newly every year, and it is annual dead
Die about 3,000,000.Currently, the mainly efficiently degeneration-resistant biography of AIDS clinical treatment records virus therapy, the therapy is not only effective
It controls HIV to replicate, and the immune function of AIDS patient can be rebuild, the door of hope is opened for the treatment of AIDS.People once posted
Wish in by the fully erased internal HIV of HAART, to achieve the purpose that thoroughly to cure AIDS.But it is subsequent it was verified that
Although HAART can inhibit patient's body virus replication to the maximum extent, plasma viral load (virus load) is made to be reduced to
The level that existing common detection methods do not measure, but still with the presence of viral persistence in the infected's body, once stop drug therapy, disease
Level (Ho, D.D.Toward HIV eradication or remission malicious carrying capacity can rebound to treatment again before:the
Tasks ahead.Science, 1998.280:1866-1867.).HIV is difficult to be completely removed in vivo important original
It can hide in the memory CD4+T cells of tranquillization because being HIV-1, which is by sub-fraction HIV infection
It activating CD4+T cell transformations and generates, the provirus integrated lacks transcriptional activity, thus, by immune system and it will not resist
The drug attack of retrovirus.Although infected individuals carry latent infected cells negligible amounts, attenuation rate is so
Slowly, so that being intended within the individual survival phase that HAART treatments is only leaned on to be thoroughly removed to be impossible.Therefore, HIV latent infections
Tranquillization CD4+T cells be constitute body inner virus bunker (reservoir) major part, while be also current clinic control
The huge obstacle of HIV cannot be thoroughly removed by treating.It is generally acknowledged that molecular mechanism and integration site that HIV-1 latent infected cells are formed
The chromatin state at place, the presence of inhibition nucleosome nuc-1, the epigenetic modification of representative, host transcription are turned to acetyl
The factors such as the factor such as NF- κ B and virus transcription activation factor Tat are related.Mechanism accordingly has researched and proposed the latent disease of removing
The therapeutic strategy of malicious repository attempts to express by the provirus of drug-induced HIV latent infected cells, makes its latent virus
It activates again, in combination with efficiently degeneration-resistant biography record virus therapy and under human immune system effect, to kill the latent sense of activation
The cell of dye accelerates removing (the Richman et al.The Challenge of Finding a Cure of virus base with this
For HIV Infection, Science, 2009,1304,323).Although the strategy clinically has several treatment sides
Case, its result are still not up to expectations, and are not that activator is invalid, though it is exactly effectively but its toxic side effect is big, and at home, so far still
There is not the anti-AIDS new drug of independent intellectual property right to list.Thus, research and development are with independent intellectual property right, safe and effective, inexpensive
The novel intervention drug that can remove HIV-1 virus repositories have a very important significance.
Rabies are most ancient one of infectious diseases, are a kind of caused by rabies viruses (Rabies virus, RABV)
Infectious diseases common to human beings and animals.Rabies are the highest acute infectious diseases of mankind's case fatality rate so far, once morbidity, case fatality rate is almost
Up to 100%.According to the World Health Organization (WHO) report, the whole world has about 55000 people to die of rabies every year, wherein most
It is distributed in Africa and Asia.In recent years, there are about 3000 people or so to die of rabies every year on average in China, and majority is happened at vast agriculture
Village area or the combination area of city and country are mainly infected by the malicious animal bite of band.RABV enters position of biting with saliva, in flesh
Enter nearest nerve fibre in meat after of short duration incubation, and gradually migrated to brain, wound location is closer from brain, and morbidity is also got over
Soon.Once into nervous system, RABV is replicated and migration will be not easy to interfere.It is to prevent and control mad dog that (PEP) is disposed after exposure
The most effective approach of disease, according to the degree bitten, three kinds of means of generally use：1) 0.1% suds is used to rinse wound；2)
Wound is injected with anti-RABV immunoglobulins, neutralizes the virus of wound location；3) urgent immunity 4-5 needles rabies vaccine.Currently, being permitted
More people only pay attention to immune rabies vacciness, have ignored the processing of wound, RABV is made to generate enough neutralizing antibodies in vaccine immunity
(it is generally necessary to 2-4 weeks) slips into brain before, causes immuning failure.Since anti-RABV immunoglobulins are at high price, it is difficult to wide
The general processing for wound, and need the medical institutions to certain scale that could receive anti-RABV immunoglobulins processing, it is difficult
Wound is handled with the first time after biting.Therefore, there is an urgent need to develop one kind effectively inhibiting RABV in wound site,
And it is cheap, it can be widely applied for the drug of vast medical low developed area.
It is more than 1.8 hundred million people that Hepatitis C Virus (HCV), which has infected the whole world,.It is estimated that annual new infection is to 400
Ten thousand people, wherein 70% would develop into chronic hepatitis.In developed country, 50-76%s and all liver of the HCV to whole liver cancer cases
2/3rds of transplanting are responsible.Standard treatment polyethylene glycol a joint Ribavirins (nucleoside medicine) are only in 50-60%
Patient in work and have significant side effect related.Therefore, there is an urgent need to new HCV drugs.
There are about the carrier that 1.3 hundred million people are hepatitis B at present in China, that is to say, that hepatitis B surface antigen (Hbs Ag) is in
The ratio that existing positive carrier accounts for total number of persons is up to 10.34%.Probably there is the hepatitis carrier of a quarter can be final
Develop into chronic hepatitis, hepatic sclerosis, primary carcinoma of liver.In addition to this, about there be more than 1,200 ten thousand chronic hepatitis disease in the whole nation
People dies of liver patient about 300,000 people every year in these patients, wherein it is about 150,000 people to die of liver cancer.Worse feelings
Condition is that the gravid woman of about 40% carrying hepatitis B surface antigen (Hbs Ag) can pass through this side of vertical transmission
Formula so that newborn's (ratio is about 80-90%) of 80-90 ten thousand becomes the carrier of chronic hepatitis B virus, not only serious
Human health is threatened, and gives family, society brings heavy financial burden.Treating chronic hepatitis B, mainly use it is antiviral,
Immunological regulation improves the complex treatments such as liver function and anti-hepatic fibrosis, but antiviral therapy is wherein main, crucial treatment
Measure.The drug of present anti-hepatitis virus (HBV), which continuously emerges, to be generated, especially nucleoside medicine (nucleoside
Analogues), become the hot spot of antiviral drugs research in recent years, the very fast progress of acquirement.
Zika virus (Zika virus) is one kind mainly by her mosquito-borne carapuru virus.The virus is most earlier than 1947
Year accidentally found in the rhesus macaque of stockaded village of Uganda card jungle by yellow fever monitoring network, then in nineteen fifty-two in Uganda and
It is found in Tanzania crowd.On May 7th, 2015, Pan American Health Organization and the World Health Organization announce that Brazilian health authority is true
It accepts the prevalence of Brazilian northeast zika virus, stockaded village's card is mainly popular in Africa, Southeast Asia and Pacific Islands before this,
However, since in May, 2015, the multiple countries in America, which start report, the propagation of stockaded village's card, and predicts that other have stockaded village card medium mosquito
The country of worm will all have Case report successively.Then less than in year, by March 10, World Health Organization's report
There are the countries and regions of local transmission case to have increased to 52.By on March 11st, 2016, China had 13 input venereal diseases
Example.Currently, the treatment for zika virus uses aspirin and nonsteroidal and-inflammatory drug more, but stockaded village's card and dengue fever are deposited
The possibility infected at the same time, the above drug may increase the risk of dengue hemorrhagic.It is known that zika virus category flavivirus
Section, Flavivirus, it is spherical in shape, it is single strand plus RNA virus, diameter 40-70nm, it is about 10.8kb to have coating, length, and coding is big
A amino acid about more than 3400.The resistance of zika virus is unknown, but the virus of Flavivirus is not general acidproof, thermo-labile.NS3 eggs
White enzyme is that the proteolysis of its polyprotein is processed so that necessary to virus replication, inhibits virus to increase to make NS3pro become
The attractive therapy target grown, the NS2B-NS3 proteinase complex of zika virus is in viral entire life cycle
In play crucial adjustment effect, after only the proteinase complex is activated and completes a series of hydrolysis, virus could start
Reproduction process.Therefore, the non-structural protein NS2B-NS3 proteinase complex of zika virus also become one it is crucial antiviral
Drug targets, so the non-structural protein enzyme for zika virus carries out inhibitor screening to the relevant medicament research and development of zika virus
It has a very big significance.
Invention content
The invention solves one side technical problem：By 3 '-deoxidations -3 ', 4 '-two dehydrogenation nucleoside compounds are for controlling
Treat or prevent the host that rabies viruses, inhibition of HIV, HCV virus, HBV viruses, zika virus infect.By 3 '-deoxidations -3 ', 4 ' -
Two dehydrogenation nucleoside compounds are their corresponding triphosphoric acid shapes in host cell enzymes metabolic conversion with the prodrug of phosplate
Formula.The nucleoside polymerase inhibitor of the triphosphate forms simulates natural nucleotide, with their naturally occurring ribonucleoside triphosphotes
Competition extends for RNA or DNA, to by being used as the chain terminator of chain terminator or delay come suppressing virus replication.
According to an aspect of the invention, there is provided a kind of 3 '-deoxidations -3 ' of formula (I), 4 '-two dehydrogenation ucleosides chemical combination
Object treats or prevents the side of rabies viruses, inhibition of HIV, HCV virus, HBV viruses, the application of zika virus and preparation formula (I)
Method.
Wherein：
B is selected from adenine and derivativeGuanine and derivativeCytimidineThymidineWherein,
X is independent to be selected from：
R1And R2It is independent to be selected from：C1-3Alkyl, C2-6Alkenyl, C2-6Alkynyl, C1-6Alkoxy, halogen, C1-6Halogenated alkyl,
C1-6Amide groups ,-NHSO3C1-6Alkyl, nitro, cyano；
Z is selected from：
(1) it is H, OH, halogen, C1-20Alkyl, C3-6Naphthenic base, C1-6Halogenated alkyl, aryl or heteroaryl.
(2)Wherein
Y is selected from O or S；
R1And R2It is independent to be selected from：
(a)OR3, wherein R3It is H, C1-20Alkyl, C3-6Naphthenic base, C1-6Halogenated alkyl, aryl or heteroaryl, including but not
It is limited to the phenyl optionally replaced by 1-3 substituent group, the 1-3 substituent group is independent to be selected from：C1-3Alkyl, C2-6Alkenyl,
C2-6Alkynyl, C1-6Alkoxy, halogen, C1-6Halogenated alkyl, C1-6Amide groups ,-NHSO3C1-6Alkyl, nitro, cyano, (CH2)1- 6CO2R4、-N(R4)2、-SO2N(R4) 2 and COR5；
R4It is independently H or C1-6Alkyl；
R5It is-OR4Or-N (R4)2。
(b)
Wherein, n 0-6,
R6It is H, C1-3Alkyl；
R7It is independent to be selected from H, C1-20Alkyl, C3-6Naphthenic base, C1-6Halogenated alkyl, aryl or heteroaryl, including but it is unlimited
In the phenyl optionally replaced by 1-3 substituent group, the 1-3 substituent group is independent to be selected from：C1-3Alkyl, C2-6Alkenyl, C2-6
Alkynyl, C1-6Alkoxy, halogen, C1-6Halogenated alkyl.
The preparation route is：
The method basic step is：
It is compound 3 '-deoxidation -3 ', 4 '-two dehydrogenations -1 ', 2 ', 5 '-triacetyl ribose (II) and protection or free
Base is catalyzed to obtain 3 '-deoxidations -3 ' with titanium tetrachloride, 4 '-two dehydrogenations -2 ', 5 '-diacetyl ribonucleotide (III), then through ammonia
Methanol solution hydrolyzes to obtain 3 '-deoxidations -3 ', 4 '-two dehydrogenations-ribonucleotide (IV), then with PYCl3(wherein, Y is O or S) reaction
Alcoholysis afterwards obtains compound 3 '-deoxidation -3 ', 4 '-two phosphate-based ribonucleotide of-O- of dehydrogenation -5 ' (I).
In the wherein described route,
Base B is selected from adenine and derivativeGuanine and derivativeCytimidineThymidine
Wherein, X is independent is selected from：C1-3Alkyl, C2-6Alkenyl, C2-6Alkynyl, C1-6Alkoxy, halogen, C1-6Halogenated alkyl,
C1-6Amide groups ,-NHSO3C1-6Alkyl, nitro, cyano；
R1And R2It is independent to be selected from：
(a)OR3, wherein R3It is H, C1-20Alkyl, C3-6Naphthenic base, C1-6Halogenated alkyl, aryl or heteroaryl, including but
It is not limited to the phenyl optionally replaced by 1-3 substituent group, the 1-3 substituent group is independent to be selected from：C1-3Alkane
Base,
C2-6Alkenyl, C2-6Alkynyl, C1-6Alkoxy, halogen, C1-6Halogenated alkyl, C1-6Amide groups ,-NHSO3C1-6Alkyl,
Nitro, cyano, (CH2)1-6CO2R4、-N(R4)2、-SO2N(R4) 2 and COR5；
R4It is independently H or C1-6Alkyl；
R5It is-OR4Or-N (R4)2。
According to another aspect of the present invention, it is to provide formula (I) compound and is inhibiting rabies viruses, inhibition of HIV, HCV diseases
Poison, HBV viruses, the in vitro activity test of zika virus and data.
Specific implementation mode
Embodiment 1
3 '-deoxidations -3 ', 4 '-two-O- phosphate cytidines of dehydrogenation -5 ' (compound 4a-e)
Its preparation route is as follows：
Intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenations -2 ', the preparation of 5 '-diacetyl cytidines (compound 2)
Cytimidine (11.0g, 0.1mol) is suspended in 180mL dry toluenes, 15mL anhydrous DMFs, hexamethyl two is added
Silicon amine alkane 20mL, is heated to reflux 4h, and system is clarified completely, is cooled to 70 DEG C, evaporated under reduced pressure.3 '-deoxidations -3 ' are added at room temperature,
4 '-two dehydrogenations -1 ', 2 ', 5 '-triacetyl ribose (25.8g, 0.1mol), anhydrous methylene chloride 200mL, titanium tetrachloride 1.0mL,
5h to be reacted at room temperature, is filtered, filtrate washing, organic phase is dried with anhydrous sodium sulfate, and filtering and concentrating obtains middle body compound 2,
25.2g, yield 85.2%.
Intermediate 3 '-deoxidation -3 ', the preparation of 4 '-two dehydrogenations-cytidine (compound 3)
Compound 2 (15.0g, 0.05mol) is added in the methanol solution 100mL of 10% ammonia, stirs 2h at room temperature, reacted
It finishes, is concentrated under reduced pressure, ethyl alcohol is added in residue, heating is completely dissolved, filtering, stirs lower cooling crystallization.Filtering, it is dry, in vain
Color solid, i.e. intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenations-cytidine (compound 3), 7.52g, yield 75.3%.
The preparation of 3 '-deoxidations -3 ', 4 '-two dehydrogenase 35s '-diethyl phosphate cytidine (compound 4a)
Phosphorus oxychloride (5.0g, 0.033mol) is dissolved in triethyl phosphate 50mL, 0 DEG C of addition above compound 3
(7.4g, 0.033mol) reacts 2h, and after the reaction was complete, filtering obtains white solid and is added into absolute ethyl alcohol 50mL, 60 DEG C
4h is reacted, the reaction was complete, and ethyl alcohol is recovered under reduced pressure.Silica gel column chromatography after purification, obtains off-white powder, i.e. compound 3 '-deoxidation-
3 ', 4 '-two dehydrogenase 35 '-O- diethyl phosphate cytidines, 5.0g, yield 50.5%.
Compound 4a：ESI m/z 362[M+1].1H NMR (600MHz, DMSO), δ：9.01 (1H, d), 8.10 (2H, s,
NH2), 5.82 (1H, d), 5.21 (1H, d), 4.82 (1H, dd), 4.75 (1H, m), 4.66 (2H, s), 4.07 (4H, q), 1.26
(6H, t).
4b, 4c, 4d, 4e can be prepared with the method for preparing 4a.
Embodiment 2
3 '-deoxidations -3 ', 4 '-two-O- phosphate ribothymidines of dehydrogenation -5 ' (compound 7a-e)
Its preparation route is as follows：
Intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenations -2 ', the preparation of 5 '-diacetyl ribothymidines (compound 5)
Anhydrous methylene chloride 200mL is added in thymidine (12.60g, 0.1mol), 3 '-deoxidations -3 ' are added at room temperature,
4 '-two dehydrogenations -1 ', 2 ', 5 '-triacetyl ribose (25.6g, 0.1mol), titanium tetrachloride 1.0mL reacts 5h at room temperature, filtering,
Filtrate is washed, and organic phase is dried with anhydrous sodium sulfate, and filtering and concentrating obtains middle body compound 5,24.1g, yield 81.5%.
Intermediate 3 '-deoxidation -3 ', the preparation of 4 '-two dehydrogenation ribothymidines (compound 6)
Compound 5 (16.10g, 0.05mol) is added in the methanol solution 100mL of 10% ammonia, stirs 2h at room temperature, instead
It should finish, be concentrated under reduced pressure, ethyl alcohol is added in residue, heating is completely dissolved, filtering, stirs lower cooling crystallization.Filtering, it is dry,
White solid, i.e. intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenation ribothymidines (compound 6), 8.42g, yield 70.1%.
3 '-deoxidations -3 ', the preparation of 4 '-two-O- diethyl phosphate ribothymidines of dehydrogenation -5 ' (compound 7a)
Phosphorus oxychloride (5.00g, 0.033mol) is dissolved in triethyl phosphate 50mL, 0 DEG C of addition above compound 6
(8.00g, 0.033mol) reacts 3h, and after the reaction was complete, filtering obtains white solid and is added into absolute ethyl alcohol 50mL, 60 DEG C
4h is reacted, the reaction was complete, and ethyl alcohol is recovered under reduced pressure.Silica gel column chromatography after purification, obtains off-white powder, i.e. compound 3 '-deoxidation-
3 ', 4 '-two-O- diethyl phosphate ribothymidines of dehydrogenation -5 ', 4.87g, yield 40.1%.
Compound 7a：ESI m/z 377[M+1].1H NMR (600MHz, DMSO), δ：9.81 (1H, s, NH), 7.43
(1H, d), 6.12 (1H, d), 4.86 (1H, m), 4.78 (1H, dd), 4.60 (2H, d), 4.01 (4H, q), 2.38 (3H, s),
1.24 (6H, t).
7b, 7c, 7d, 7e can be prepared with the method for preparing 7a.
Embodiment 3
3 '-deoxidations -3 ', 4 '-two-O- phosphate ribose adenosines of dehydrogenation -5 ' (compound 10a-e)
Intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenations -2 ', the preparation of 5 '-diacetyl ribose adenosines (compound 8)
Adenine (13.7g, 0.1mol) is suspended in 250mL dry toluenes, 20mL anhydrous DMFs, hexamethyl two is added
Silicon amine alkane 20mL, is heated to reflux 7h, and system is clarified completely, is cooled to 70 DEG C, evaporated under reduced pressure.3 '-deoxidations -3 ' are added at room temperature,
4 '-two dehydrogenations -1 ', 2 ', 5 '-triacetyl ribose (25.8g, 0.1mol), anhydrous methylene chloride 200mL, titanium tetrachloride 1.0mL,
5h to be reacted at room temperature, is filtered, filtrate washing, organic phase is dried with anhydrous sodium sulfate, and filtering and concentrating obtains middle body compound 8,
25.5g, yield 76.8%.
Intermediate 3 '-deoxidation -3 ', the preparation of 4 '-two dehydrogenation ribose adenosines (compound 9)
Compound 8 (16.67g, 0.05mol) is added in the methanol solution 100mL of 10% ammonia, stirs 2h at room temperature, instead
It should finish, be concentrated under reduced pressure, ethyl alcohol is added in residue, heating is completely dissolved, filtering, stirs lower cooling crystallization.Filtering, it is dry,
White solid, i.e. intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenation ribose adenosines (compound 9), 7.47g, yield 60.1%.
3 '-deoxidations -3 ', the preparation of 4 '-two-O- diethyl phosphate ribose adenosines of dehydrogenation -5 ' (compound 10a)
Phosphorus oxychloride (3.77g, 0.025mol) is dissolved in triethyl phosphate 50mL, 0 DEG C of addition above compound 9
(6.27g, 0.025mol) reacts 4h, and after the reaction was complete, filtering obtains white solid and is added into absolute ethyl alcohol 50mL, 60 DEG C
6h is reacted, the reaction was complete, and ethyl alcohol is recovered under reduced pressure.Silica gel column chromatography after purification, obtains off-white powder, i.e. compound 3 '-deoxidation-
3 ', 4 '-two-O- diethyl phosphate ribose adenosines of dehydrogenation -5 ', 4.93g, yield 51.3%.
Compound 10a：ESI m/z 386[M+1].1H NMR (600MHz, DMSO), δ：8.37 (1H, s), 8.21 (1H,
S), 7.12 (2H, s, NH2), 5.76 (1H, d), 4.80 (1H, d), 4.45 (2H, s), 4.22 (1H, dd), 3.88 (4H, q),
1.23 (6H, t).
10b, 10c, 10d, 10e can be prepared with the method for preparing 10a.
Embodiment 4
3 '-deoxidations -3 ', 4 '-two-O- phosphate ribose guanosines of dehydrogenation -5 ' (compound 13a-e)
Its preparation route is as follows：
Intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenations -2 ', the preparation of 5 '-diacetyl ribose guanosines (compound 11)
Adenine (15.3g, 0.1mol) is suspended in 250mL dry toluenes, 20mL anhydrous DMFs, hexamethyl two is added
Silicon amine alkane 20mL, is heated to reflux 7h, and system is clarified completely, is cooled to 70 DEG C, evaporated under reduced pressure.3 '-deoxidations -3 ' are added at room temperature,
4 '-two dehydrogenations -1 ', 2 ', 5 '-triacetyl ribose (25.8g, 0.1mol), anhydrous methylene chloride 200mL, titanium tetrachloride 1.0mL,
5h to be reacted at room temperature, is filtered, filtrate washing, organic phase is dried with anhydrous sodium sulfate, and filtering and concentrating obtains middle body compound 11,
24.5g, yield 70.2%.
Intermediate 3 '-deoxidation -3 ', the preparation of 4 '-two dehydrogenation ribose guanosines (compound 12)
Compound 11 (17.55g, 0.05mol) is added in the methanol solution 100mL of 10% ammonia, stirs 2h at room temperature, instead
It should finish, be concentrated under reduced pressure, ethyl alcohol is added in residue, heating is completely dissolved, filtering, stirs lower cooling crystallization.Filtering, it is dry,
White solid, i.e. intermediate 3 '-deoxidation -3 ', 4 '-two dehydrogenation ribose guanosines (compound 9), 10.69g, yield 80.5%.
3 '-deoxidations -3 ', the preparation of 4 '-two-O- diethyl phosphate ribose guanosines of dehydrogenation -5 ' (compound 13a)
Phosphorus oxychloride (3.77g, 0.025mol) is dissolved in triethyl phosphate 50mL, 0 DEG C of addition above compound 12
(6.67g, 0.025mol) reacts 5h, and after the reaction was complete, filtering obtains white solid and is added into absolute ethyl alcohol 60mL, 60 DEG C
5h is reacted, the reaction was complete, and ethyl alcohol is recovered under reduced pressure.Silica gel column chromatography after purification, obtains off-white powder, i.e. compound 3 '-deoxidation-
3 ', 4 '-two-O- diethyl phosphate ribose guanosines of dehydrogenation -5 ', 4.27g, yield 42.7%.
Compound 13a：ESI m/z 402[M+1].1H NMR (600MHz, DMSO), δ：8.20(2H,s,NH2), 8.0
(1H, s, NH), 7.51 (1H, s), 6.22 (1H, d), 4.80 (1H, d), 4.50 (2H, s), 4.04 (1H, dd), 3.98 (4H, q),
1.23 (6H, t).
13b, 13c, 13d, 13e can be prepared with the method for preparing 13a.
Embodiment 5
3 '-deoxidations -3 ', the external Anti-HIV-1 Active experiment (mtt assay) of 4 '-two-O- phosphate nucleosides of dehydrogenation -5 '
Test procedure:
(1) by 3 '-deoxidations -3 ' of various concentration to be tested, 4 '-two-O- phosphate nucleosides medicine samples of dehydrogenation -5 ' are in 96 holes
1640 culture medium doubling dilution is used in cell plates, each hole is made to retain 100 μ L liquids, and 50 μ L (about 10000) MT- is added to each hole
4 cells add 50 μ L 100TCID50HIV-1 virus liquids.It sets normal cell controls simultaneously and virus-infected controls is several
Hole, if drug AZT positive controls.
(2) 37 DEG C, 5%CO2Cell incubator culture 48h.Third day, 2 times of concentration 1640 liquid, 20 μ L of each hole benefit continued to cultivate
48h。
(3) fixed, dyeing：Gently 200 μ L of fixer are added with regard to culture medium in reject, and room temperature fixes 5min；PBS is gently washed
It washs cell 2 times, dyeing liquor (MTT) 100 μ L is added, be incubated 50min；Microscopic counting locus coeruleus.Virus is calculated according to following formula
Inhibiting rate is fitted test data using software, acquires half-inhibition concentration (IC50).The experimental results are shown inthe following table.
Inhibiting rate=[1- (experimental group locus coeruleus number-cell controls group locus coeruleus number)/(virus control group locus coeruleus number-cell controls
Group locus coeruleus number)] * 100%.
The activity test in vitro result of table 1 anti-HIV-1 virus
Experimental result：
Test result shows 3 '-deoxidations -3 ', 4 '-two-O- phosphate nucleosides of dehydrogenation -5 ', which part reactive compound
IC50Higher than a line inverase AZT, other 3 '-deoxidations -3 ', 4 '-two-O- phosphate nucleosides of dehydrogenation -5 ' also show that compared with
The strong external effect for inhibiting HIV.
Embodiment 6
3 '-deoxidations -3 ', the external anti-HCV activity experiment of 4 '-two-O- phosphate nucleosides of dehydrogenation -5 '
Step：
(1) in 96 orifice plates, the good Huh7.5.1 cells of growth conditions are added, it is (about 10000 thin per about 100 μ L of hole
Born of the same parents), suitable DMEM culture mediums are added, are placed in 37 DEG C, 5% CO2In incubator, culture is for 24 hours.
(2) it is added with DMSO gradient dilutions (10 times) 3 '-deoxidations -3 ' to be measured, 4 '-two-O- phosphate nucleosides of dehydrogenation -5 '
6 gradients are arranged in compound and control drug RBV (Ribavirin), each compound, and each gradient repeats three holes, is arranged simultaneously
Blank control wells and negative control hole, it is the every holes 200 μ L to add culture medium to final volume.37 DEG C are placed in, 5% CO2In incubator
Culture.
(3) 72h is cultivated, the WST-1 solution of 20 μ L is added per hole, 96 well culture plates are placed in 5% CO2, 37 DEG C of incubations
2h.96 orifice plates are placed on shaking table and are shaken one minute, to mix well system to be detected, in measuring OD in microplate reader450Value.
(4) it utilizes software to handle, calculates untested compound to the growth inhibition ratio of Huh7.5.1 cells, acquire to be measuredization
Close the IC of object50Value.
Growth inhibition ratio=[1- (experimental port OD values/negative control OD value)] * 100%.
The activity test in vitro result of table 2 HCV-Ab IgG virus
Experimental result：
Test result shows that, relative to Ribavirin, 3 '-deoxidations -3 ', 4 '-two-O- phosphate nucleosides of dehydrogenation -5 ' are shown
Go out relatively strong external inhibition HCV virus activity.
Embodiment 7
3 '-deoxidations -3 ', the external Anti-HBV effect experiment of 4 '-two-O- phosphate nucleosides of dehydrogenation -5 '
Step：
(1) one bottle of Hep G 2.2.15 cells for covering with culture bottle are taken, are made after being digested with 0.25% pancreatin unicellular outstanding
Liquid adjusts cell concentration to 1 × 10 after counting4A m L-1, are inoculated in 96 porocyte culture plates, per 100 μ L of hole.It will training
Foster plate is placed in CO2In, 37 DEG C, 5.0%CO2Under the conditions of cultivate for 24 hours.
(2) after cells grow up to the individual layer is separately added into the 3 '-deoxidations -3 ' containing various concentration, 4 '-two-O- of dehydrogenation -5 '
1640 culture mediums of phosphate nucleosides, while Lamivudine positive controls and blank control group are set, the 3 multiple holes trainings of each concentration
After supporting 5 days.
(3) it is added the MTT 20 μ L containing crystal violet into every hole, 37 DEG C, 5.0%CO2Under the conditions of be incubated 4h, discard supernatant
After liquid, 200 μ L of DMSO are added into every hole, is placed in micro oscillator, gently shakes to crystal violet and almost dissolve, uses
168-1000XC types microplate reader is in each hole OD values of 490nm (reference wavelength 630nm) wavelength detecting.
Inhibiting rate=[(blank control wells OD values-dosing holes OD values)/blank control wells OD values] * 100%
The activity test in vitro result of 3 resisting HBV virus of table
Experimental result：
Test result shows that relative to Ribavirin, 3 '-deoxidations -3 ', 4 '-two dehydrogenation nucleosides show relatively strong external suppression
HBV virus activities processed.
Embodiment 8
3 '-deoxidations -3 ', the external rabies cytotoxic activity experiment of 4 '-two dehydrogenations-nucleosides
Step：
1, the medicine sample of various concentration to be tested is used into 1640 culture medium doubling dilution in 96 porocyte plates, each hole is made to retain
100 μ L liquids are added 50 μ L (about 10000) to each hole and grow up to single layer SK-N-SH cells cell after pancreatin digests, add
50μL 100TCID50Rabies virus strain (SAD) virus liquid.Set normal cell controls and virus-infected controls several holes simultaneously.
2,37 DEG C, 5%CO2Cell incubator culture 48h.Third day, 2 times of concentration 1640 liquid, 20 μ L of each hole benefit continued to cultivate
48h。
3, fixed, dyeing：Gently 200 μ L of fixer are added with regard to culture medium in reject, and room temperature fixes 5min；PBS is gently washed
Dyeing liquor (MTT) 100 μ L are added in cell 2 times, are incubated 50min；Microscopic counting locus coeruleus.Virus suppression is calculated according to following formula
Rate processed is fitted test data using software, acquires half-inhibition concentration (IC50).The experimental results are shown inthe following table.
Inhibiting rate=[1- (experimental group locus coeruleus number-cell controls group locus coeruleus number)/(virus control group locus coeruleus number-cell controls
Group locus coeruleus number)] * 100%.
The activity test in vitro result of 4 rabies poison
Experimental result：
Test result shows that 3 '-deoxidations -3 ', 4 '-two dehydrogenation nucleosides show relatively strong external inhibition rabies viruses (SAD)
The activity of virus.
Embodiment 9
3 '-deoxidations -3 ', the external inhibition zika virus activity test of 4 '-two dehydrogenations-nucleosides
1, various concentration medicine sample to be tested is used into 1640 culture medium doubling dilution in 96 porocyte plates, each hole is made to retain
100 μ L liquids are added 50 μ L (about 10000) to each hole and grow up to single layer African green monkey kidney cell (Vero) after pancreatin digests,
Add 50 μ L 100TCID50Zika virus liquid.Set normal cell controls and virus-infected controls several holes simultaneously.
2,37 DEG C, 5%CO2Cell incubator culture 48h.Third day, 2 times of concentration 1640 liquid, 20 μ L of each hole benefit continued to cultivate
48h。
3, fixed, dyeing：Gently 200 μ L of fixer are added with regard to culture medium in reject, and room temperature fixes 5min；PBS is gently washed
Dyeing liquor (MTT) 100 μ L are added in cell 2 times, are incubated 50min；Microscopic counting locus coeruleus.Virus suppression is calculated according to following formula
Rate processed is fitted test data using software, acquires half-inhibition concentration (IC50).The experimental results are shown inthe following table.
Inhibiting rate=[1- (experimental group locus coeruleus number-cell controls group locus coeruleus number)/(virus control group locus coeruleus number-cell controls
Group locus coeruleus number)] * 100%.
The activity test in vitro result of 5 anti-zika virus of table
Experimental result：
Test result shows that 3 '-deoxidations -3 ', 4 '-two dehydrogenation nucleoside compounds show relatively strong external inhibition stockaded village's card disease
The activity of poison.
The present invention relates to nucleoside compound antivirus technology field, specifically 3 '-deoxidation -3 ', 4 '-two dehydrogenation ucleosides
Compound is inhibiting HIV-1, rabies viruses, HCV, HBV, stockaded village's card disease to show higher activity, shows 3 '-deoxidations -3 ', and 4 ' -
Two dehydrogenation nucleoside compounds have very big application potential in anti-virus aspect.
Claims (6)
1. a kind of bis- dehydrogenation nucleoside compound of 3'- deoxidations -3', 4'-, which is characterized in that its chemical general formula is (I)：
Wherein：
B is selected from adenine and derivativeGuanine and derivativeCytimidineThymidine
Wherein, X is independent is selected from R1And R2It is independent to be selected from：C1-3Alkyl, C2-6Alkenyl, C2-6Alkynyl, C1-6Alkoxy, halogen,
C1-6Halogenated alkyl, C1-6Amide groups ,-NHSO3C1-6Alkyl, nitro, cyano；
Z is selected from：
(1) it is H, OH, halogen, C1-20Alkyl, C3-6Naphthenic base, C1-6Halogenated alkyl, aryl or heteroaryl.
(2)Wherein
Y is selected from O or S；
R1And R2It is independent to be selected from：
(a)OR3, wherein R3It is H, C1-20Alkyl, C3-6Naphthenic base, C1-6Halogenated alkyl, aryl or heteroaryl, including but not limited to
The phenyl optionally replaced by 1-3 substituent group, the 1-3 substituent group is independent to be selected from：C1-3Alkyl, C2-6Alkenyl, C2-6Alkynes
Base, C1-6Alkoxy, halogen, C1-6Halogenated alkyl, C1-6Amide groups ,-NHSO3C1-6Alkyl, nitro, cyano, (CH2)1-6CO2R4、-
N(R4)2、-SO2N(R4) 2 and COR5；
R4It is independently H or C1-6Alkyl；
R5It is-OR4Or-N (R4)2。
(b)
Wherein, n 0-6,
R6It is H, C1-3Alkyl；
R7It is independent to be selected from H, C1-20Alkyl, C3-6Naphthenic base, C1-6Halogenated alkyl, aryl or heteroaryl, including but not limited to appoint
The phenyl that selection of land is replaced by 1-3 substituent group, the 1-3 substituent group is independent to be selected from：C1-3Alkyl, C2-6Alkenyl, C2-6Alkynes
Base, C1-6Alkoxy, halogen, C1-6Halogenated alkyl.
2. application of the bis- dehydrogenation nucleoside compound of 3'- deoxidations -3', 4'- described in claim 1 in antiviral.
3. 3'- deoxidations -3', 4'- bis- dehydrogenations-nucleoside compound described in claim 1 is used to prepare the application of drug, described
Drug is used to treat by the host of HIV-1 or HIV-2 infection.
4. 3'- deoxidations -3', 4'- bis- dehydrogenations-nucleoside compound described in claim 1 is used to prepare the application of drug, described
Drug is for treating by the host of rabies virus infection.
5. 3'- deoxidations -3', 4'- bis- dehydrogenations-nucleoside compound described in claim 1 is used to prepare the application of drug, described
Drug is used to treat by the host of hepatitis C (HCV), hepatitis B (HBV) virus infection.
6. 3'- deoxidations -3', 4'- bis- dehydrogenations-nucleoside compound described in claim 1 is used to prepare the application of drug, described
Drug is for treating the host infected by zika virus.
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| CN114773417A (en) * | 2022-04-06 | 2022-07-22 | 郑州大学 | Cordycepin phosphate and preparation method and application thereof |
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| Title |
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| MAGDALENA PETROVÁ，等: "Straightforward synthesis of 3"-deoxy-3",4"-didehydronucleoside-5"-aldehydes via 2",3"-O-orthoester group elimination: a simple route to 3",4"-didehydronucleosides", 《TETRAHEDRON LETTERS》 * |
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| WO2022260535A1 (en) * | 2021-06-11 | 2022-12-15 | Victoria Link Limited | Antiviral nucleoside analogues |
| CN114773417A (en) * | 2022-04-06 | 2022-07-22 | 郑州大学 | Cordycepin phosphate and preparation method and application thereof |
| CN114773417B (en) * | 2022-04-06 | 2023-08-22 | 郑州大学 | Cordycepin phosphate and preparation method and application thereof |
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