CN107028955A - Purposes of 25 hydroxy cholesterols in flavivirus is suppressed - Google Patents
Purposes of 25 hydroxy cholesterols in flavivirus is suppressed Download PDFInfo
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- 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
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/575—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of three or more carbon atoms, e.g. cholane, cholestane, ergosterol, sitosterol
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- 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
Abstract
The present invention relates to chemical medicine field, a kind of purposes of 25 hydroxy cholesterol in flavivirus is suppressed is disclosed.Specifically, 25HC is disclosed to prepare for the medicine for preventing and/or treating flaviviridae infections, suppression flavivirus, preparing for preventing and/or treating the application in the medicine of microcephaly caused by zika virus infection.The invention also discloses a kind of pharmaceutical composition, contain 25HC, at least one assistant medicament for being conducive to suppressing flavivirus, and at least one pharmaceutically acceptable carrier.By using 25HC, the present invention can suppress flavivirus, and host is played a protective role, specifically, and 25HC can suppress flaviviridae infections in people's cell, mouse and macaque model, and can prevent and/or treat microcephaly caused by zika virus.
Description
Technical field
The present invention relates to chemical medicine field, in particular it relates to a kind of use of 25-HYDROXY CHOLESTEROL in flavivirus is suppressed
On the way.
Background technology
Zika virus (Zika Virus, ZIKV), because with mosquito matchmaker, Asymptomatic Carriers and sex track transmission, spread speed
Very fast, the health care belt given people is threatened.Zika virus is felt for one in nineteen forty-seven from stockaded village's card jungle of Uganda at first
It is isolated in the rhesus monkeys of dye.Nineteen fifty-two report first case people infected patient (Dick, 1952；Dick et al., 1952).It
Before, zika virus infection people only produces light symptoms such as arthralgia, weak, fash and conjunctivitis.Zika virus occurs within 2015
In Brazil with unexpected congenital abnormality great outburst such as microcephaly, Guillain Barre syndrome and meningitis
(Araujo et al., 2016；Cao-Lormeau et al., 2016；Carteaux et al., 2016；Musso and Gubler, 2016).
It is now recognized that these serious Manifestations of nervous system be by pregnant woman's intrauterine infection zika virus so as to accidentally occur microcephaly, nature
Intrauterine growth restriction caused by miscarriage and placental insufficiency.Discovery zika virus infecting mouse testis is had been reported that recently
After can cause male sterility.Therefore, zika virus infection causes huge challenge to Global Health system.
Unfortunately, currently without approval anti-zika virus medicine or vaccine it is various caused by zika virus to protect
Disease.Zika virus can be replicated efficiently in the mouse after type interferon receptors knockout, point out interferon activated gene
(ISGs) infection level (Aliota et al., 2016 can be reduced；Lazear et al., 2016).Afterwards, researcher has found that host is thin
Born of the same parents' control zika virus infection is realized by interferon signal path, and host can protect place by producing substantial amounts of ISGs
Chief cell is not infected (Bayer et al., 2016；Hamel et al., 2015；Quicke et al., 2016).There is experiment
Confirm that IFITM1 and IFITM3 in the early stage in zika virus life cycle, ISGs families can suppress viral infection.
Other IFITM3 can suppress the Apoptosis (Savidis et al., 2016) of zika virus induction.But, in innate immunity confrontation
What is played a key effect in zika virus course of infection removes IFITM1, IFITM3 (interferon-induced
Transmembrane 1and 3proteins, interferon-induced transmembrane protein) outer other ISGs effect it is also indefinite.
Cholesterol -25- hydroxylases (Cholesterol-25-Hydroxylase) can be catalyzed cholesterol and be transformed into 25- hydroxyls
Base cholesterol (25-Hydroxycholesterol, 25HC), 25HC is the oxidized form steroidal alcohol of the naturally occurring in human body
(Holmes et al., 2011；Janowski et al., 1999；Kandutsch et al., 1978), its answering in terms of flavivirus is suppressed
With there is not been reported.
The content of the invention
The purpose of the present invention is to overcome the deficiencies in the prior art there is provided a kind of 25HC new application, that is, is suppressing flavivirus
In purposes.
It was found by the inventors of the present invention that 25HC can effectively suppress the duplication of flavivirus, therefore, first in vivo and in vitro
Aspect, the application in being used to prevent and/or treat the medicine of flaviviridae infections is being prepared the invention provides 25HC.
Second aspect, the invention provides applications of the 25HC in flavivirus is suppressed.
The third aspect, is used to preventing and/or treating small caused by zika virus infection the invention provides 25HC in preparation
Application in the medicine of head disease.
Fourth aspect, the invention provides a kind of pharmaceutical composition, it is favourable that the pharmaceutical composition contains 25HC, at least one
In the assistant medicament for suppressing flavivirus, and at least one pharmaceutically acceptable carrier.
5th aspect, the invention provides a kind of 25-HYDROXY CHOLESTEROL, the 25-HYDROXY CHOLESTEROL is used to prevent and/or control
Flaviviridae infections are treated, suppresses flavivirus or prevents and/or treat microcephaly caused by zika virus infection.
By using 25HC, the present invention can suppress flavivirus, and host is played a protective role, specifically, and 25HC can be
Suppress flaviviridae infections in people's cell, mouse and macaque model, and can prevent and/or treat microcephaly caused by zika virus
Disease.
Other features and advantages of the present invention will be described in detail in subsequent embodiment part.
Brief description of the drawings
Accompanying drawing is, for providing a further understanding of the present invention, and to constitute a part for specification, with following tool
Body embodiment is used to explain the present invention together, but is not construed as limiting the invention.In the accompanying drawings：
Fig. 1 is that 25HC can effectively suppress the duplication of zika virus in vitro, wherein, Figure 1A surveys for 25HC cytotoxicity
Test result, Figure 1B is inhibitions of the 25HC to zika virus that plaque assay detects various concentrations, and Fig. 1 C detect for RT-qPCR
As a result, Fig. 1 D are the result figure of Immunofluorescence test zika virus E protein, wherein, NITD008 is the suppression of wide spectrum anti-flavivirus
Agent；
Fig. 2 is inhibition figures of the 25HC to other flavivirus, wherein, Fig. 2A, 2B, 2C and 2D are respectively various concentrations
25HC is imitated to zika virus (ZIKV), dengue virus (DENV), yellow fever virus (YFV) and the suppression of West Nile Virus (WNV)
Fruit is schemed；
Fig. 3 is the result figure that 25HC suppresses zika virus duplication in Mice Body, wherein, Fig. 3 A are that 25HC is small in BALB/c
To the inhibition result figure of zika virus in mouse body, Fig. 3 B are the survival of the A129 mouse of injection 25HC postoperative infection zika viruses
Situation result figure, Fig. 3 C-3E are respectively to inject after 25HC (successive administration 7 days), infect the A129 mouse of zika virus the 3rd
My god, the 7th day detection viremia virusemia result figure and the virus titer result figure in the 7th day detection mouse brain；
Fig. 4 is inhibition figures of the 25HC in rhesus macaque to zika virus, wherein, Fig. 4 A are the signal of experimentation
Figure, titre testing result viral in the rhesus macaque blood for injection 25HC postoperative infection zika viruses Fig. 4 B, Fig. 4 C are injection 25HC
Viral titre testing result in the rhesus macaque urine of postoperative infection zika virus；
Fig. 5 is protecting effects of the 25HC to microcephaly caused by zika virus, wherein, Fig. 5 A are tire mouse brain immunofluorescence
The statistical result of detection, Fig. 5 B are the result figure that tire mouse brain virus load is analyzed with RT-qPCR, and Fig. 5 C are tire mouse brain rip cutting
Face Nissl coloration result figures, Fig. 5 D are the result figure that each cortex in tire mouse brain cross section is dyed with Nissl, and Fig. 5 E are tire mouse brain
Portion NeuN+, Tbr1+The result figure of skin thickness, Fig. 5 F are P-H3 testing results, and Fig. 5 G are Caspase3 expression quantity testing results；
Fig. 6 is safety evaluatios of the 25HC to pregnant mouse and suckling mouse, wherein, Fig. 6 A are that pregnant mouse detects knot in period from prenatal to postnatal body weight
Really, Fig. 6 B are that suckling mouse growth conditions observe result figure, and Fig. 6 C are first day suckling mouse body weight testing result after birth, and Fig. 6 D are injection
ALT Activity determination results in 25HC A129 mice serums, Fig. 6 E are concentration of the 25HC in A129 mice serums anaplasia at any time
Change situation map.
Embodiment
The embodiment to the present invention is described in detail below.It should be appreciated that described herein specific
Embodiment is merely to illustrate and explain the present invention, and is not intended to limit the invention.
The end points and any value of disclosed scope are not limited to the accurate scope or value herein, these scopes or
Value should be understood to comprising the value close to these scopes or value.For number range, between the endpoint value of each scope, respectively
It can be combined with each other between the endpoint value of individual scope and single point value, and individually between point value and obtain one or more
New number range, these number ranges should be considered as specific open herein.
In the present invention, in the case where not making opposite explanation, the term used such as " suppression flavivirus " typically refers to suppression
The duplication of manufacture-yellow virus, so as to reduce the titre of flavivirus.
The present invention relates to applications of the 25HC in preventing and/or treating flaviviridae infections.Meanwhile, the invention further relates to one kind
Prevention and/or the method for the treatment of flaviviridae infections, it includes (effective dose) 25HC being administered to subject.More specifically, this
Invention is related to 25HC and is preparing the application in being used to prevent and/or treat the medicine of flaviviridae infections.
Suppress the application in flavivirus the invention further relates to 25HC (in vivo or in vitro), particularly suppress jaundice in vitro
Application in poison.Correspondingly, the invention further relates to a kind of method for suppressing flavivirus, it is included (effective dose) 25HC and Huang
Viruses contact (25HC is such as administered to subject, or be administered to arthropod that the carrier of flavivirus sucks blood (mosquito, tick,
Sand fly etc.), or it is administered to other vessel that there may be flavivirus).
The invention further relates to applications of the 25HC in preventing and/or treating microcephaly caused by zika virus infection.Accordingly
Ground, the invention further relates to a kind of method prevented and/or treat microcephaly caused by zika virus infection, it includes will (effectively
Amount) 25HC is administered to subject (particularly female subject, more preferably gestational period subject).More specifically, the present invention is related to
And 25HC is preparing the application in being used to preventing and/or treating the medicine of microcephaly caused by zika virus infection.
Above-mentioned flavivirus (flavivirus) can be various common by the arthropod-borne sucked blood for this area
Cause the tunicary single positive chain RNA virus of tool of infection, preferably zika virus, dengue virus, yellow fever virus and West Nile
At least one of virus.
Wherein, above-mentioned subject can be the common easy mammal by flaviviridae infections, particularly primate
(such as people or monkey) or rodent (such as mouse).
Wherein, the method for administration can be conventional mode, be such as injected intraperitoneally or be injected intravenously.The dosage of administration can be with
For the conventional dosage (effective dose) in this area, can according to various parameters, the age in particular according to subject, body weight and sex come
It is determined that.For example, for female, 3-4 week old, body weight 15-20g BALB/c mouse, 25HC consumption can be 25-100mg/kg
Body weight (intraperitoneal injection), preferably 50-75mg/kg body weight.For 3 age body weight 4kg rhesus macaque, 25HC consumption can be
2.5-10mg/kg body weight (intravenous injection), preferably 5-7.5mg/kg body weight.
25HC can be prepared into different formulations by those skilled in the art according to the concrete mode of administration.Therefore, it is another
Aspect, the invention further relates to a kind of pharmaceutical composition, the pharmaceutical composition contains 25HC, at least one and is conducive to suppressing flavivirus
Assistant medicament, and at least one pharmaceutically acceptable carrier.The assistant medicament can be interferon and/or rope fluorine cloth
Wei, etc..The pharmaceutically acceptable carrier can be to be present in any composition in pharmaceutical composition and inactive ingredients,
Therefore including diluent, adhesive, lubricant, disintegrant, filler, colouring agent, wetting agent, emulsifying agent, pH buffer, preservative
Deng.
The invention further relates to a kind of 25HC, as it was previously stated, it is used to prevent and/or treats flaviviridae infections, suppresses jaundice
Microcephaly caused by poison or prevention and/or treatment zika virus infection.
The present invention will be described in detail by way of examples below.In following examples, room temperature refers to " 20 DEG C "；25HC is purchased
From Sigma companies, article No. is H1015；GZ01 plants of zika virus and other flavivirus are stored in Military Medical Science Institute's Virus seed library；
Vero cells are purchased from ATCC companies, and article No. is CCL-81；BHK-21 cells are purchased from ATCC companies, and article No. is CCL-10；Hela is thin
Born of the same parents are purchased from ATCC companies, and article No. is CCL-2；BALB/c mouse comes from Shanghai purchased from Jackson Laboratory, A129 mouse
Pasteur institute, mouse breeds by Military Medical Science Institute's Experimental Animal Center；ICR mouse are purchased from the magnificent company of dimension tonneau；Henghe
Monkey is purchased from Military Medical Science Institute's Experimental Animal Center, 3 years old, 4kg；All animal experiments are dynamic in Military Medical Science Institute's experiment
Thing service centre completes, and obtains Ethics Committee of Military Medical Science Institute (IACUC-13-2016-001) license.
Experimental method used in following embodiments is conventional method unless otherwise specified.Institute in following embodiments
Material, reagent etc., unless otherwise specified, can be obtained from commercial channels.
Embodiment 1
(1) 25HC can effectively suppress the duplication of zika virus in vitro.
With the 25HC of various concentrations pretreatment Vero cells 36 hours, etoh solvent (EtOH) was as negative control, then
Toxicity of the 25HC to cell is detected with MTT experiment.As a result as shown in Figure 1A：10 μM of 25HC is to Vero cells without toxicity
(Figure 1A).
In order to detect inhibitions of the 25HC to zika virus, Vero cells are pre-processed 12 hours with the 25HC of various concentrations
Afterwards, with GZ01 plants of infection cells (MOI 0.1) of zika virus.After 48 hours, the supernatant of cell is taken to do plaque assay, or take thin
Cellular lysate liquid does RT-qPCR detection virus titers, and solvent EtOH is used as control.Comprise the following steps that：
(1) plaque assay：Paving 2 × 105Individual BHK-21 cells are in night incubation in 12 orifice plates.Then PBS cell is used
Once, with cell conditioned medium sample incubation to be detected 1 hour；Then cell is containing 1% low melting-point agarose and containing 2%FBS
DMEM culture mediums in be incubated again 4 days.With being counted after violet staining to plaque.
(2) RNA separation, reverse transcription and real-time quantitative PCR：With Purelink RNA extracts kits (Thermo
Fisher the total serum IgE of cell, tissue or virus) is extracted.With OneStep PrimeScript RT-PCR kits (Takara,
Beijing), pass through RT-qRCR technology for detection viral RNA duplication amounts (similarly hereinafter).Forward primer used：5’-
GGTCAGCGTCCTCTCTAATAAACG-3’(SEQ ID NO：1)；Reverse primer：5’-GCACCCTAGTGTCCACTTTTTCC-
3’(SEQ ID NO：2)；Probe：5’-FAM-AGCCATGACCGACACCACACCGT-BQ1-3’(SEQ ID NO：3).
Respectively as shown in figs. ib and 1 c, 0.4 μM of 25HC can be effectively for plaque assay and RT-qPCR testing results
Suppress the infection of zika virus.
Finally, Hela cells are pre-processed 12 hours with 5 μM of 25HC, then with GZ01 plants of infection cell (MOI of zika virus
0.1).After 48 hours, with the amount of Immunofluorescence test zika virus E protein, (antibody is 4G2, ATCC：HB-112), wherein, make
With solvent EtOH as negative control, the inhibitor NITD008 of wide spectrum anti-flavivirus is used as positive control.As a result show, 25HC
Infection (Fig. 1 D) of the zika virus in Hela cells can effectively be suppressed.
(2) inhibitions of the 25HC to other flavivirus (including dengue virus, yellow fever virus and West Nile Virus).
25HC is have detected with RT-qPCR method to other flavivirus (including dengue virus, yellow fever virus and West Nile
Virus) inhibition.That is, after pre-processing Vero cells 12 hours with the 25HC of various concentrations, zika virus GZ01 is used respectively
Strain, the strain of 2-43 plants of dengue virus, yellow fever virus 17D and chin-01 plants of infection cells (MOI 0.1) of West Nile Virus.48 is small
Shi Hou, takes cell pyrolysis liquid to do real-time quantitative PCR detection virus titer.As a result show, 25HC suppresses zika virus, Dengue disease
Poison, yellow fever virus and the IC50 of West Nile Virus are respectively 188nM, 406nM, 526nM and 1109nM (such as Fig. 2A -2D institutes
Show), illustrate that 25HC has wide spectrum inhibition to flavivirus.
(3) 25HC can suppress the duplication of zika virus in Mice Body.
With 25HC (50mg/kg) or equivalent solvent (Vehicle, H β CD, hydroxypropyl cyclodextrin) to BALB/c mouse or
A129 mouse (female mice of 3-4 weeks size) are pre-processed, then with GZ01 plants (10 of zika virus5PFU/ mouse) abdominal cavity note
Penetrate and infect these mouse.After infection 12 hours, these mouse continue intraperitoneal injection 25HC (50mg/kg) or solvent 1 day daily
(BALB/c) or up to 7 days (A129 mouse).The symptom and death condition of observation mouse were until 21 days after infection daily.Use RT-
QPCR detects the zika virus infecting mouse of solvent or 25HC processing viral RNA amount in serum or brain after 4 days and 7 days.Use Log-
Rank (Mantel-Cox) examines the survival condition for comparing solvent or the treated zika virus infecting mouses of 25HC.As a result show
Show, 25HC can effectively suppress duplication (Fig. 3 A, 3C-3D) of the zika virus in BALB/c or A129 mouse, and can be small in A129
Dead mouse (Fig. 3 B) effectively caused by protection zika virus in mouse；25HC can effectively suppress duplication viral in mouse brain simultaneously
(Fig. 3 E).
(4) inhibitions of the 25HC in rhesus macaque to zika virus.
Rhesus macaque progress is injected intravenously with 25HC (1.5mg/kg) (n=2) or solvent EtOH (Control, n=3) pre-
Processing, by GZ01 plants (10 of zika virus after 1 day3PFU/ monkey) intramuscular injection is in rhesus monkeys.After infection 6 hours, by perseverance
River monkey continues intravenous injection 25HC (5mg/kg) daily or solvent up to 7 days (concrete operations flow is shown in Fig. 4 A).Monkey is observed daily
Symptom, and take blood, urinate, viral RNA amount in serum or urine is detected with RT-qPCR.As a result show, 25HC can effectively reduce stockaded village's card
Virus titre (Fig. 4 B-4C) viral in rhesus macaque blood and urine.
(5) protecting effects of the 25HC to microcephaly caused by zika virus.
The zika virus set up with laboratory causes the model (referring to Li et al., 2016) of microcephaly in mouse, detection
Protecting effects of the 25HC to microcephaly.To ICR mouse peritoneal injections 25HC (50mg/kg) or solvent control before virus infection
(Vehicle, H β CD, hydroxypropyl cyclodextrin) carries out pretreatment 6 hours.To the μ l zika viruses of ICR right side of mice intracerebroventricular 1
SZ01 plants (6.5 × 105PFU/ml), second half tire mouse injection of culture medium is used as control.After 6 hours, pregnant mouse continues daily injection
25HC or solvent.After 5 days, take tire mouse brain to carry out immunofluorescence or dyeing detection, operate as follows：
Immunofluorescence dyeing and co-focusing imaging are quantitative：For frozen section, tissue is fixed in 4% paraformaldehyde,
Then dehydration and the freezing in tissue freezing liquid in 30% sucrose.Thickness is 40 μm of histotomies in 10% horse serum, 0.3%
Room temperature is closed 1 hour in Triton X-100 and PBS containing 5%BSA.With the primary antibody Phospho-Histone3 of following component
(P-H3)(CST,9701S,1:1000),Activated-caspase3(CST,9664s,1:500),Tbr1(Abcam,
ab31940,1:500) and from the serum (1 for infecting zika virus patient:100) it is incubated overnight.The control of zika virus
Serum (1：100) Healthy People is come from, while compareing mouse serum (1：500).Nucleus is with DAPI (Thermo Fisher)
Dyeing.Section is placed on LSM 700 (Carl Zeiss) Laser Scanning Confocal Microscope and taken pictures, while image Imaris and ImageJ
Analyzed.All data are all analyzed with Prism softwares (GraphPad).Examined with unpaired t and carry out statistical analysis.
The data gone out are average ± SEM.As a result it is as shown in Figure 5.
As indicated by figures 5 a-5b, find that 25HC can effectively reduce tire mouse brain with immunofluorescence and RT-qPCR detections respectively
The content of virus.Dyed and found with Nissl, it is thinning that 25HC can be effectively protected cerebral cortex caused by zika virus infection
Change with the ventricles of the brain is big (Fig. 5 C).For each subregion of cortex, Nissl coloration results are displayed that, 25HC can effectively prevent by
Cortex subregion is thinning caused by zika virus infection, as shown in Figure 5 D, wherein, MZ:Marginal belt (marginal zone)；CP:
Cortical plate (cortical plate)；SP:Bottom plate (sub-plate)；IZ:Intermediate Gray (intermediate zone).Especially
SP regions, the tire mouse brain infected in zika virus can't detect, and be able to detect that in the tire mouse brain of 25HC processing
SP regions (Fig. 5 D-5E).
Pass through the marker molecules NeuN and the marker molecules of immature neuron of Immunofluorescence test mature neuron
Tbr1, it is found that 25HC can also improve NeuN/Tbr1 ratio, can improve the development of neuron and ripe level, protect by stockaded village
Neuronal development caused by card virus infection is obstructed (Fig. 5 E).By detecting neuronal precursor (neuronal
Progenitor cells, NPC) propagation mark P-H3, this is the mark that a NPC is in the meiosis M phases.As a result
Display zika virus infection can reduce amounts of the P-H3 in NPC, and 25HC can recover this phenomenon, improve P-H3 expression quantity
(Fig. 5 F).Other zika virus infection can cause Caspase3 expression quantity to increase, along with the enhancing of the apoptosis of cell.And 25HC
The expression of Caspase3 albumen can be reduced, so as to reduce the generation (Fig. 5 G) of Apoptosis.
(6) safety evaluatios of the 25HC to pregnant mouse and suckling mouse
In order to evaluate securities of the 25HC in pregnant mouse and common Mice Body, in ICR pregnant mouse E14.5 days, intraperitoneal injection
25HC (50mg/kg), solvent control (Vehicle, H β CD), successive administration 5 days, and the changes of weight of the pregnant mouse of Continuous Observation, and
Whether miscarriage or premature labor phenomenon are had.As shown in Figure 6A, injection 25HC group and the pregnant mouse changes of weight trend of solvent group suppress, and
And all do not miscarry or premature labor, 3 weeks postpartum body weight all returns to normal level.The suckling mouse body weight of birth and development are all unaffected
(Fig. 6 B-6C).
Meanwhile, influences of the 25HC to transaminase in serum (ALT) activity is also have detected, as a result as shown in Figure 6 D, in A129
Successive administration is after 7 days in mouse, and serum alt activity and control group are not significantly different.Illustrate under the dosage, 25HC is to pregnant mouse
And normal mouse is safe.25HC concentration is raised to highest in 4 hours upon administration in serum, gradually reduces afterwards, small to 24
Shi Hou, 25HC concentration are reduced to normal level (Fig. 6 E).Illustrate that the 25HC of injection crosses metabolism in blood back warp and can return to physiology
Level.
Can be seen that from above-mentioned experimental result can effectively suppress flavivirus using 25HC, so as to for prevent and/or
Treat flaviviridae infections.
The preferred embodiment of the present invention described in detail above, still, the present invention are not limited in above-mentioned embodiment
Detail, in the range of the technology design of the present invention, a variety of simple variants can be carried out to technical scheme, this
A little simple variants belong to protection scope of the present invention.
It is further to note that each particular technique feature described in above-mentioned embodiment, in not lance
In the case of shield, can be combined by any suitable means, in order to avoid unnecessary repetition, the present invention to it is various can
The combination of energy no longer separately illustrates.
In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally
The thought of invention, it should equally be considered as content disclosed in this invention.
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SEQUENCE LISTING
<110>Suzhou system medicine research institute
Inst. of Epidemiology and Microbiology, Academy of Military Medical Sciences, PL
Inst. of Genetics and Development Biology, CAS
<120>Purposes of the 25-HYDROXY CHOLESTEROL in flavivirus is suppressed
<130> I43495PUMC
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> The sequence is synthesized
<400> 1
ggtcagcgtc ctctctaata aacg 24
<210> 2
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> The sequence is synthesized
<400> 2
gcaccctagt gtccactttt tcc 23
<210> 3
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> The sequence is synthesized
<400> 3
agccatgacc gacaccacac cgt 23
Claims (6)
1.25- hydroxy cholesterols are preparing the application in being used to prevent and/or treat the medicine of flaviviridae infections.
Application of the 2.25- hydroxy cholesterols in flavivirus is suppressed, particularly suppresses the application in flavivirus in vitro.
3.25- hydroxy cholesterols are in the medicine for being used for preventing and/or treat the caused microcephaly of zika virus infection is prepared
Using.
4. the application according to any one in claim 1-3, wherein, the flavivirus be zika virus, dengue virus,
At least one of yellow fever virus and West Nile Virus.
5. a kind of pharmaceutical composition, it is characterised in that the pharmaceutical composition contains 25-HYDROXY CHOLESTEROL, at least one and is conducive to
Suppress the assistant medicament of flavivirus, and at least one pharmaceutically acceptable carrier.
6. a kind of 25-HYDROXY CHOLESTEROL, it is characterised in that the 25-HYDROXY CHOLESTEROL is used to prevent and/or treat flavivirus sense
Microcephaly caused by dye, suppression flavivirus or prevention and/or treatment zika virus infection.
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| CN201710044968.8A CN107028955B (en) | 2017-01-20 | 2017-01-20 | Purposes of the 25-HYDROXY CHOLESTEROL in the drug that preparation inhibits flavivirus |
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| CN107028955B CN107028955B (en) | 2019-08-16 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021089721A1 (en) * | 2019-11-05 | 2021-05-14 | Hercules Pharmaceuticals B.V. | Treatment, amelioration or prevention of a viral infection. |
| CN113018421A (en) * | 2020-03-12 | 2021-06-25 | 中国人民解放军军事科学院军事医学研究院 | Use of cholesterol-25-hydroxylase and enzymatic products thereof for the preparation of a medicament for inhibiting a novel coronavirus |
| CN113082208A (en) * | 2019-12-23 | 2021-07-09 | 中国科学院分子细胞科学卓越创新中心 | Medicine for blocking microbial infection, reducing cholesterol and preventing and treating related tumors and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104998275A (en) * | 2015-06-25 | 2015-10-28 | 中国科学院广州生物医药与健康研究院 | Function of CH25H and 25-OHC in immune response and application thereof as immunomodulator in vaccines |
| CN105362278A (en) * | 2014-09-01 | 2016-03-02 | 苏州系统医学研究所 | Preparation containing 25-hydroxycholesterol and preparation method thereof and anti-virus application |
-
2017
- 2017-01-20 CN CN201710044968.8A patent/CN107028955B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105362278A (en) * | 2014-09-01 | 2016-03-02 | 苏州系统医学研究所 | Preparation containing 25-hydroxycholesterol and preparation method thereof and anti-virus application |
| CN104998275A (en) * | 2015-06-25 | 2015-10-28 | 中国科学院广州生物医药与健康研究院 | Function of CH25H and 25-OHC in immune response and application thereof as immunomodulator in vaccines |
Non-Patent Citations (1)
| Title |
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| SU-YANG LIU 等: "Interferon-Inducible Cholesterol-25-Hydroxylase Broadly Inhibits Viral Entry by Production of 25-Hydroxycholesterol", 《IMMUNITY》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021089721A1 (en) * | 2019-11-05 | 2021-05-14 | Hercules Pharmaceuticals B.V. | Treatment, amelioration or prevention of a viral infection. |
| EP3827824A1 (en) * | 2019-11-26 | 2021-06-02 | Hercules Pharmaceuticals B.V. | Treatment of congenital zika virus syndrome |
| CN113082208A (en) * | 2019-12-23 | 2021-07-09 | 中国科学院分子细胞科学卓越创新中心 | Medicine for blocking microbial infection, reducing cholesterol and preventing and treating related tumors and application thereof |
| CN113082208B (en) * | 2019-12-23 | 2022-07-08 | 中国科学院分子细胞科学卓越创新中心 | Medicine for blocking microbial infection, reducing cholesterol and preventing and treating related tumors and application thereof |
| CN113018421A (en) * | 2020-03-12 | 2021-06-25 | 中国人民解放军军事科学院军事医学研究院 | Use of cholesterol-25-hydroxylase and enzymatic products thereof for the preparation of a medicament for inhibiting a novel coronavirus |
| CN113018421B (en) * | 2020-03-12 | 2022-05-10 | 中国人民解放军军事科学院军事医学研究院 | Use of cholesterol-25-hydroxylase and enzymatic product thereof in the manufacture of a medicament for inhibiting a novel coronavirus |
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|---|---|
| CN107028955B (en) | 2019-08-16 |
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