CN106085868A - One strain produces aspergillosis and the application thereof of HIV (human immunodeficiency virus)-resistant activity material - Google Patents
One strain produces aspergillosis and the application thereof of HIV (human immunodeficiency virus)-resistant activity material Download PDFInfo
- Publication number
- CN106085868A CN106085868A CN201610424586.3A CN201610424586A CN106085868A CN 106085868 A CN106085868 A CN 106085868A CN 201610424586 A CN201610424586 A CN 201610424586A CN 106085868 A CN106085868 A CN 106085868A
- Authority
- CN
- China
- Prior art keywords
- formula
- hiv
- methanol
- compound shown
- aspergillosis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/12—Ketones
- A61K31/122—Ketones having the oxygen directly attached to a ring, e.g. quinones, vitamin K1, anthralin
-
- 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
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/351—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom not condensed with another ring
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P15/00—Preparation of compounds containing at least three condensed carbocyclic rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/66—Aspergillus
Abstract
The invention discloses a strain and produce aspergillosis and the application thereof of HIV (human immunodeficiency virus)-resistant activity material.The bacterial strain number of this aspergillosis is CPCC400735, and it is at the numbered CGMCC No.12376 that registers on the books at China Committee for Culture Collection of Microorganisms's common micro-organisms center.The methanolic extract of aspergillosis CPCC400735 fermentation culture medium suppression ratio to HIV 1 when concentration is 4mg/mL is 99.9%.Compound shown in formula 1 to 4 when concentration is 100 μMs to the suppression ratio of HIV 1 all more than 96%, IC50It is worth 2.4 32.6 μMs.Compound shown in the methanolic extract of the aspergillosis CPCC400735 fermentation culture medium of the present invention and formula 1 to formula 4 has stronger inhibitory activity to HIV 1, and toxicity is less.
Description
Technical field
The present invention relates to a strain and produce aspergillosis and the application thereof of HIV (human immunodeficiency virus)-resistant activity material.
Background technology
HIV (Human Immunodeficiency Virus) virus, Chinese full name HIV (human immunodeficiency virus), belong to inverse
The one of Retroviral.After inhibition of HIV invades host cell, meeting heavy damage host body immunologic function, thus cause serious
Opportunistic infection so that causing the secondary immunodeficiency being characterized with malignant disease, be acquired immune deficiency syndrome (AIDS)
(acquired immunodeficiency syndrome, AIDS).Patient With Aids case was found in the U.S. in 1981,
Then begin to propagate the most rapidly, the most one of significant threat becoming human health.Inhibition of HIV can be divided into
HIV-1 type and HIV-2 type, both genomes have the biggest difference, and wherein HIV-1 is the master causing global spread of aids
Want pathogen.
Chemotherapy remains currently the most important ones HIV-1 treatment means, has 26 kinds of chemical entities to enter clinic so far
Application.Classify by its mechanism of action, these marketed drug are mainly reverse transcriptase inhibitors and protease inhibitor, add up
To 22 kinds.And only 4 kinds of other mechanism of action, including 2 kinds of integrase inhibitors and 2 kinds of viral entry inhibitors.Along with anti-
The continuous research and development application of HIV medicine, particularly relies on Cocktail treatment mode to treat HIV sufferers, can have
Effect reduces virus load, notable prolongation survival of patients time, but the problems such as the compliance of drug resistance, untoward reaction and patient are still
Often cause the failure of clinical treatment.Therefore, it is still necessary to the new anti-HIV-1 medicines of Persisting exploitation is for clinical practice.
Microbe species is huge, has the metabolite of enriched types, is find high-efficiency low-toxicity HIV (human immunodeficiency virus)-resistant activity material one
Individual important sources.The bacterial strain of HIV (human immunodeficiency virus)-resistant activity material is produced, then from its metabolite by setting up screening model rapid screening
Find that to have the compound of HIV (human immunodeficiency virus)-resistant activity or have the compound of certain activity be guide, carry out structure simplification and modification, disclose
The mechanism of action and structure activity relationship, be the effective way finding new inverase.
Summary of the invention
The technical problem to be solved is how to suppress HIV.
In order to solve above technical problem, the invention provides a strain aspergillosis.
Aspergillosis provided by the present invention (Aspergillus sp.), its bacterial strain number is that CPCC400735 is (also known as CPCC
400735), it is at the numbered CGMCC that registers on the books at China Committee for Culture Collection of Microorganisms's common micro-organisms center
No.12376。
Above-mentioned aspergillosis (Aspergillus sp.) CPCC400735 can with conidium, mycelia or containing conidium and/
Or presented in the mycelium of mycelia.
Above-mentioned aspergillosis (Aspergillus sp.) CPCC400735 application in preparing hiv inhibitor, above-mentioned aspergillosis
(Aspergillus sp.) CPCC400735 treats or/and prevent answering in acquired immune deficiency syndrome (AIDS) medicine in preparation
With, the hiv inhibitor that obtained by above-mentioned aspergillosis (Aspergillus sp.) CPCC400735, by above-mentioned aspergillosis
The treatment that (Aspergillus sp.) CPCC400735 prepares is or/and prevention acquired immune deficiency syndrome (AIDS) medicine is equal
Belong to protection scope of the present invention.
The culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC400735 falls within protection scope of the present invention.
The culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC400735 is by above-mentioned aspergillosis (Aspergillus
Sp.) material in CPCC400735 cultivates the culture vessel obtained in microbiological culture media.
In the culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC400735, described microbiological culture media can be bent
Mould culture medium (can cultivate the culture medium of aspergillosis), and it can be solid medium, semisolid culturemedium or fluid medium.Described micro-
Biological medium can be rice culture medium, potato dextrose agar or other funguses well known to those skilled in the art
Fermentation medium etc..In the culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC 400735, described rice culture medium can
Be made up of rice and water, it is possible to be made up of rice, water and inorganic salt, it is possible to be made up of rice, water and nitrogen source, it is possible to by rice,
Water, nitrogen source and inorganic salt are made.Described potato dextrose agar can be made up of Rhizoma Solani tuber osi, glucose, agar and water,
Also can be made up of Rhizoma Solani tuber osi, glucose, agar, water and inorganic salt, it is possible to be made up of Rhizoma Solani tuber osi, carbon source, agar, water and nitrogen source,
Also can be made up of Rhizoma Solani tuber osi, carbon source, agar, nitrogen source and inorganic salt.Other fungi fermentations well known to those skilled in the art described
Culture medium can by a certain or several quick-acting, imitate carbon source, a certain or several effect nitrogen source quick-acting, slow, water and/or inorganic salt etc. late
Make.
Above, described rice can be rice or brown rice.Rice or brown rice are all the goods of Oryza glutinosa.Oryza glutinosa refers to not go
Except the fruit of the husk of paddy rice, Oryza glutinosa is made up of rice husk, peel, seed coat, perisperm, aleurone, endosperm and embryo.Brown rice refers to Oryza glutinosa
Slough rice husk, retain the goods of other each several part of Oryza glutinosa；Rice refers to only retain endosperm, and is all sloughed by Oryza glutinosa remainder
Goods.Carbon source is growth of microorganism one class nutrient, is carbon compound, including saccharide, oils and fats, organic acid and organic acid esters
Quick-acting with small molecular alcohol etc., imitate carbon source late.Nitrogen source refers to provide the material of nitrogen element needed for microbial nutrition, including peanut cake
The effect nitrogen sources quick-acting, slow such as powder, soybean cake powder, yeast powder, peptone, ammonia, ammonium salt and nitrate.
In the culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC400735, the temperature of described cultivation can be 20-30
DEG C, incubation time can be 10-40 days.
Present invention also offers a kind of hiv inhibitor.
The active component of a kind of hiv inhibitor provided by the present invention is from above-mentioned aspergillosis (Aspergillus with methanol
Sp.) culture of CPCC400735 extracts the material being dissolved in methanol obtained.
Present invention also offers the preparation method of this hiv inhibitor, including using methanol from above-mentioned aspergillosis (Aspergillus
Sp.) culture of CPCC400735 extracts obtain being dissolved in the material of methanol, using described material as the activity of hiv inhibitor
Composition.
Present invention also offers a kind for the treatment of or/and prevent acquired immune deficiency syndrome (AIDS) medicine.
Treatment provided by the present invention is or/and the active component of prevention acquired immune deficiency syndrome (AIDS) medicine is to use methanol
The material being dissolved in methanol obtained is extracted from the culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC400735.
Present invention also offers this treatment or/and prevent the preparation method of acquired immune deficiency syndrome (AIDS) medicine, including
Extract from the culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC400735 with methanol and obtain being dissolved in the material of methanol,
Using described material as treatment or/and prevent the active component of acquired immune deficiency syndrome (AIDS) medicine.
Present invention also offers another kind of hiv inhibitor.
The active component of another kind of hiv inhibitor provided by the present invention, is the compound shown in formula II:
Described R is
Present invention also offers the method preparing hiv inhibitor, including using methanol from above-mentioned aspergillosis (Aspergillus
Sp.) culture of CPCC400735 extracts obtain being dissolved in the material of methanol, then extract from described material and obtain shown in formula II
Compound, using the compound shown in formula II as the active component of hiv inhibitor.
Present invention also offers another kind for the treatment of or/and prevent acquired immune deficiency syndrome (AIDS) medicine.
This treatment is or/and prevention acquired immune deficiency syndrome (AIDS) medicine, and its active component is the compound shown in formula II.
Present invention also offers prepare this treatment or/and prevention acquired immune deficiency syndrome (AIDS) medicine method, including
Extract from the culture of above-mentioned aspergillosis (Aspergillus sp.) CPCC400735 with methanol and obtain being dissolved in the material of methanol,
Extract from described material again and obtain the compound shown in formula II, using the compound shown in formula II as treatment or/and prevention obtains
Obtain the active component of property immunodeficiency syndrome medicine.
Present invention also offers the method preparing the compound shown in formula II.
The method of the compound prepared shown in formula II provided by the present invention, including using methanol from above-mentioned aspergillosis
The culture of (Aspergillus sp.) CPCC400735 extracts the material being dissolved in methanol obtained, then from described material
Extract and obtain the compound shown in formula II.
Present invention also offers 9)-12) method:
9) treatment or/and prevention acquired immune deficiency syndrome (AIDS) method, including to receptor use with methanol from
Above-mentioned aspergillosis (Aspergillus sp.) CPCC400735 extracts and obtains being dissolved in the material of methanol, carry out treating or/and prevent
Acquired immune deficiency syndrome (AIDS)；
10) method suppressing HIV animal, uses with methanol from above-mentioned aspergillosis including to receptor
(Aspergillus sp.) CPCC400735 culture extracts and obtains the material being dissolved in methanol to suppress HIV animal；
11) treatment is or/and the method for prevention acquired immune deficiency syndrome (AIDS), uses shown in formula II including to receptor
Compound carry out treating or/and prevent acquired immune deficiency syndrome (AIDS)；
12) method suppressing HIV animal, uses the compound shown in formula II to suppress HIV including to receptor
Infection animal.
In said method, extract from described material and obtain the compound shown in formula II and specifically can include A1)-A4) in
Corresponding steps:
A1) by described material with water-dispersible, then it is extracted with ethyl acetate, collects ethyl acetate phase, remove ethyl acetate
Ethyl acetate in mutually, obtains extractum；
A2) described extractum being carried out silica gel column chromatography separation, elution program used in silica gel column chromatography is first to use chloroform
2 column volumes of eluting；Carry out the following linear gradient elution of 13 column volumes again: flowing used is by chloroform and methanol group mutually
The mixed liquor become, in linear gradient elution flowing mutually, the volume ratio of chloroform and methanol is linearly down to 7:1 by 20:1；Finally use first again
Two column volumes of alcohol eluting；Collect the 0.9th to the 3.6th liquid that column volume elutes, by its named Fr.4-12, collect
4.5th to the 5.1st liquid that column volume elutes, by its named Fr.16-17；
A3) Fr.4-12 being carried out silica gel column chromatography, elution program used in this silica gel column chromatography is by 6 cylinders
Long-pending following linear gradient elution: the mixed liquor that flowing used is made up of petroleum ether and acetone mutually, in linear gradient elution
Flowing mutually in petroleum ether and the volume ratio of acetone be linearly down to 5:1 by 8:1；Collect the 3.2nd to the 4.5th column volume eluting
The liquid gone out, by its named Tubes 27-37；
A4) Tubes 27-37 being prepared type high performance liquid chromatography separate, this preparative high performance liquid chromatography separates institute
Filler be octadecylsilane chemically bonded silica filler, the particle diameter of filler is 5 μm, this preparative high performance liquid chromatography separate institute
The a diameter of 8mm of chromatographic column, a height of 250mm；Flowing is mutually for the mixing liquid of first alcohol and water composition, first alcohol and water in flowing mutually
Volume ratio be 4: 1；Flow velocity is 4.0mL/min, and collecting retention time is the eluting peak of 23.9min, by its named tubes
27-37-P2；Tubes 27-37-P2 is prepared type high performance liquid chromatography separate；This preparative high performance liquid chromatography separates
Filler used is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 5 μm, and this preparative high performance liquid chromatography separates
The a diameter of 8mm of chromatographic column, a height of 250mm used；Flowing mutually for first alcohol and water composition mixing liquid, flowing mutually in methanol with
The volume ratio of water is 4: 1；Flow velocity is 4.5mL/min, and collecting retention time is the eluting peak of 23.9min, and obtaining R in formula II isCompound (i.e. compound shown in formula 1)；
A5) Tubes 27-37 being prepared type high performance liquid chromatography separate, this preparative high performance liquid chromatography separates institute
Filler be octadecylsilane chemically bonded silica filler, the particle diameter of filler is 5 μm, this preparative high performance liquid chromatography separate institute
The a diameter of 8mm of chromatographic column, a height of 250mm；Flowing is mutually for the mixing liquid of first alcohol and water composition, first alcohol and water in flowing mutually
Volume ratio be 4: 1；Flow velocity is 4.0mL/min, and collecting retention time is the eluting peak of 44.6min, by its named tubes
27-37-P6；Tubes 27-37-P6 is prepared type high performance liquid chromatography separate；This preparative high performance liquid chromatography separates
Filler used is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 5 μm, and this preparative high performance liquid chromatography separates
The a diameter of 8mm of chromatographic column, a height of 250mm used；Flowing mutually for first alcohol and water composition mixing liquid, flowing mutually in methanol with
The volume ratio of water is 4: 1；Flow velocity is 5mL/min, and collecting retention time is the eluting peak of 37.7min, and obtaining R in formula II isCompound (i.e. compound shown in formula 2)；
A6) Fr.16-17 is carried out silica gel column chromatography, elution program used in this silica gel column chromatography be with by chloroform and
The chloroform of methanol composition carries out eluting with the mixing liquid of volume ratio 10:1 of methanol mutually as flowing, collects the 0th to 0.2 post
The liquid that volume elutes, by its named tubes 1-3, collects what the 0.2nd column volume to the 0.9th column volume eluted
Liquid, by its named Tubes 4-13；Carrying out opening ODS column chromatography for separation by Tubes 4-13, this opening ODS column chromatography divides
Being octadecylsilane chemically bonded silica filler from filler used, the particle diameter of filler is 50 μm, this open ODS column chromatography for separation institute
The a diameter of 4cm of chromatographic column, a height of 16cm, chromatographic column volume is 201mL；This open eluting used by ODS column chromatography for separation
Program is two step eluting, and the first step eluting mixing liquid that volume ratio is 11:9 of the first alcohol and water being made up of first alcohol and water is made
For flowing 4 column volumes of phase eluting, complete first step eluting；Second step eluting: after completing first step eluting, with methanol as stream
The dynamic eluting that carries out mutually, the 0th to the 0.5th liquid that column volume elutes that collection methanol-eluted fractions goes out, by its named Tubes4-
13-ODS-9.Tubes 4-13-ODS-9 and tubes 1-3 is merged and carries out compound-5 preparative separation shown in entitled formula 4
Preparative high performance liquid chromatography separate, the preparative high-efficient liquid phase color of this compound-5 preparative separation shown in entitled formula 4
Filler used by spectrum separation is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 5 μm, shown in this entitled formula 4
The a diameter of 8mm of chromatographic column, a height of 250mm used by preparative high performance liquid chromatography separation of compound-5 preparative separation；Flowing
Mutually for the mixing liquid of first alcohol and water composition, in flowing mutually, the volume ratio of first alcohol and water is 3: 1；Flow velocity is 4.0mL/min；Collect
Retention time is that the eluting peak of 18.2min obtains P2, collects the eluting peak that retention time is 22.2min and obtains P3, by P2 and P3
The preparative high performance liquid chromatography carrying out compound-5 preparative separation shown in described entitled formula 4 again separates, the preparative of P2
High performance liquid chromatography collects retention time in separating be that the eluting peak of 18.2min obtains R in formula II and is(i.e. compound shown in formula 4)；The preparative high performance liquid chromatography of P3 is collected in separating when retaining
Between obtain R in formula II for the eluting peak of 22.2min and beCompound (i.e. compound shown in formula 3).
Compound shown in above-mentioned formula II or the compound shown in its pharmaceutically acceptable salt, formula II or its pharmaceutically may be used
The salt application in preparing hiv inhibitor that accepts, with the compound shown in formula II or its pharmaceutically acceptable salt as activity
The hiv inhibitor of composition belongs to protection scope of the present invention.
Above, described HIV can be all the HIV of HIV-1 type, and described acquired immune deficiency syndrome (AIDS) can be drawn by HIV-1
Rise.
Above, described animal can be mammal, such as people.
Above, described hiv inhibitor and described treatment, or/and prevention acquired immune deficiency syndrome (AIDS) medicine, remove and contain
Outside active composition, also can contain suitable carrier or excipient.Here carrier material includes but not limited to water-solubility carrier
Material (such as Polyethylene Glycol, polyvinylpyrrolidone, organic acid etc.), slightly solubility carrier material are (as hard in ethyl cellulose, cholesterol
Fat acid ester etc.), enteric solubility carrier material (such as CAP and carboxylic the first and second cellulose etc.).Wherein it is preferred that water-soluble
Property carrier material.Use these materials can make multiple dosage form, include but not limited to tablet, capsule, drop pill, aerosol, ball
Agent, powder, solution, suspensoid, Emulsion, granule, liposome, transdermal agent, buccal tablet, suppository, lyophilized injectable powder etc..Permissible
It is ordinary preparation, slow releasing preparation, controlled release preparation and various particulate delivery system.In order to unit dosage forms for administration is made tablet, can
So that well known in the art various carrier is widely used.Example about carrier is, such as diluent and absorbent, such as starch, paste
Essence, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, carbamide, calcium carbonate, kaolin, microcrystalline Cellulose, aluminium silicate
Deng；Wetting agent and binding agent, as molten in water, glycerol, Polyethylene Glycol, ethanol, propanol, starch slurry, dextrin, syrup, Mel, glucose
Liquid, mucialga of arabic gummy, gelatine size, sodium carboxymethyl cellulose, lac, methylcellulose, potassium phosphate, polyvinylpyrrolidone etc.；
Disintegrating agent, such as, be dried starch, alginate, agar powder, laminaran, sodium bicarbonate and citric acid, calcium carbonate, polyoxy second
Alkene, sorbitan fatty acid ester, dodecyl sodium sulfate, methylcellulose, ethyl cellulose etc.；Disintegrate inhibitor, such as sugarcane
Sugar, glyceryl tristearate, cocoa butter, hydrogenated oil and fat etc.；Absorption enhancer, such as quaternary ammonium salt, sodium lauryl sulphate etc.；Lubrication
Agent, such as Pulvis Talci, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, Polyethylene Glycol etc..Can also be by sheet
Coated tablet, such as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablet and multilayer tablet are made in agent further.In order to incite somebody to action
Unit dosage forms for administration makes pill, and well known in the art various carrier can be widely used.Example about carrier is, the dilutest
Release agent and absorbent, such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinylpyrrolidone, Gelucire, height
Ridge soil, Pulvis Talci etc.；Binding agent such as arabic gum, Tragacanth, gelatin, ethanol, Mel, liquid sugar, rice paste or batter etc.；Disintegrate
Agent, such as agar powder, is dried starch, alginate, dodecyl sodium sulfate, methylcellulose, ethyl cellulose etc..In order to by single
Position form of administration makes suppository, and well known in the art various carrier can be widely used.Example about carrier is, the most poly-second
Glycol, lecithin, cocoa butter, higher alcohol, the ester of higher alcohol, gelatin, semi-synthetic glyceride etc..In order to by unit dosage forms for administration system
Become injection preparation, such as solution, Emulsion, lyophilized injectable powder and suspensoid, it is possible to use all diluent commonly used in the art,
Such as, water, ethanol, Polyethylene Glycol, 1,3-PD, the isooctadecanol of ethoxylation, polyoxygenated isooctadecanol, polyoxyethylene
Span etc..It addition, in order to prepare isotonic injection, can add in injection preparation appropriate sodium chloride,
Glucose or glycerol, further, it is also possible to add the cosolvent of routine, buffer agent, pH adjusting agent etc..Additionally, if desired, can also
Coloring agent, preservative, spice, correctives, sweeting agent or other material is added in pharmaceutical preparation.Use the above-mentioned dosage form can be through
Drug administration by injection, including subcutaneous injection, intravenous injection, intramuscular injection and intracavitary administration etc.；Cavity/canal drug administration, such as per rectum and vagina；
Respiratory tract administration, such as via intranasal application；Mucosa delivery.Above-mentioned route of administration preferably drug administration by injection.
It is demonstrated experimentally that the methanolic extract of aspergillosis (Aspergillus sp.) CPCC400735 fermentation culture medium is in concentration
It is 99.9% for suppression ratio to HIV-1 during 4mg/mL.The chemical combination shown in formula 1 to formula 4 separated from above-mentioned methanolic extract
Thing suppression ratio to HIV-1 when concentration is 100 μMs is respectively 99.84%, 99.99%, 98.60% and 96.82%, formula 1 to
The IC of the compound shown in formula 450Value is respectively 2.4 μMs, 4.5 μMs, 22.1 μMs and 32.6 μMs.The aspergillosis of the present invention
Compound shown in the methanolic extract of (Aspergillus sp.) CPCC400735 fermentation culture medium and formula 1 to formula 4 is to HIV-
1 has stronger inhibitory activity, and toxicity is less.
Preservation explanation
Strain name: aspergillosis (Aspergillus sp.)
Strain number: CPCC400735
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on May 5th, 2016
Register on the books numbering in preservation center: CGMCC No.12376
Accompanying drawing explanation
Fig. 1 is the structure of the compound shown in formula 4.
Fig. 2 is the structure of the compound shown in formula 2.
Fig. 3 is the structure of the compound shown in formula 1.
Fig. 4 is the structure of the compound shown in formula 3.
Fig. 5 is the compound shown in formula 41H-NMR composes.
Fig. 6 is the compound shown in formula 413C-NMR composes.
Fig. 7 is the hsqc spectrum of the compound shown in formula 4.
Fig. 8 is the HMBC spectrum of the compound shown in formula 4.
Fig. 9 is the compound shown in formula 41H-1H COSY composes.
Figure 10 is the NOESY spectrum of the compound shown in formula 4.
Figure 11 is the Rh of the compound shown in formula 42(OCOCF3)4Induction CD spectrum.
Figure 12 is the Mo (Ac) of the compound shown in formula 44Induction CD spectrum.
Figure 13 is the compound shown in formula 21H-NMR composes.
Figure 14 is the compound shown in formula 213C-NMR composes.
Figure 15 is the hsqc spectrum of the compound shown in formula 2.
Figure 16 is the HMBC spectrum of the compound shown in formula 2.
Figure 17 is the compound shown in formula 21H-1H COSY composes.
Figure 18 is the NOESY spectrum of the compound shown in formula 2.
Figure 19 is the compound shown in formula 11H-NMR composes.
Figure 20 is the compound shown in formula 113C-NMR composes
Figure 21 is the hsqc spectrum of the compound shown in formula 1.
Figure 22 is the HMBC spectrum of the compound shown in formula 1.
Figure 23 is the compound shown in formula 11H-1H COSY composes.
Figure 24 is the NOESY spectrum of the compound shown in formula 1.
Figure 25 is the relative configuration of the hexa-atomic ether ring in C-23 to the C-33 structure fragment of the compound shown in formula 1.
Figure 26 is the compound shown in formula 31H-NMR composes.
Figure 27 is the compound shown in formula 313C-NMR composes.
Figure 28 is the hsqc spectrum of the compound shown in formula 3.
Figure 29 is the HMBC spectrum of the compound shown in formula 3.
Figure 30 is the compound shown in formula 31H-1H COSY composes.
Figure 31 is the NOESY spectrum of the compound shown in formula 3.
Detailed description of the invention
Being further described in detail the present invention below in conjunction with detailed description of the invention, the embodiment be given is only for explaining
The bright present invention rather than in order to limit the scope of the present invention.Experimental technique in following embodiment, if no special instructions, is
Conventional method.Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
Potato dextrose agar (PDA) culture medium used in following embodiment: peeling potatoes, 200g is cut into little
Block, adds water boil 30min, 4 layers of filtered through gauze, adds 20g glucose, agar 17g, and distilled water constant volume, to 1000mL, boils mixing,
Sterilizing 20 minutes under the conditions of 121 DEG C, obtain PDA culture medium.
PNL4-3Luc (R-E-) in following embodiment and pHIT/G (Zhang et al.Retrovirology (2016)
13:13) public can obtain this biomaterial from NIH or applicant, this biomaterial only attach most importance to duplicate invention related experiment institute
With, can not use as other purposes.
Embodiment 1, the separation of aspergillosis (Aspergillus sp.) CPCC400735 and qualification
1.1 strains separation
Bacterial strain CPCC400735 is isolatable from the stem of Fructus Schisandrae Sphenantherae (Kadsura longipedunculata).Gather south five
Taste plant loads in valve bag, takes back laboratory treatment.Weigh 1g Fructus Schisandrae Sphenantherae stem, with 75% ethanol rinsing 30s, sterilized water
Clean 3 times, by sterile scissors, stem is shredded, put in sterilized mortar and add quartz sand and be ground.Lapping liquid is added
In centrifuge tube equipped with 9mL sterilized water, sample concentration is 10-1, after shaking up, it is made into 10 successively-2、10-3、10-4、10-5Gradient is dilute
Release liquid.Draw different gradient dilution liquid 200 μ L respectively, be respectively coated on PDA plate, be inverted in after drying up in 25 DEG C of incubators
Cultivate 3-5 days, be transferred to the fungus being separated on PDA inclined-plane cultivate, after length is good, be placed in refrigerator preservation.Take numbered
The bacterial strain of CPCC400735 carries out following qualification.
1.2 identification of strains
1.2.1 strain morphology is observed
Bacterial strain CPCC400735 Inoculating needle is chosen to PDA culture medium flat plate central authorities, 28 DEG C of incubators are cultivated.Bacterial strain
CPCC400735 is well-grown in PDA culture medium, and when cultivating 5-7 days, bacterium colony is light yellow, and quality velvet shape is to cotton-shaped, relatively
Thickness, has or does not have radial rill；Bacterium colony reverse side yellowish-brown, conidium is many or few, is just fawn, is bordering on light powder brown
Color, old after deepen, being bordering at the beginning of pink cinnamon, conidial head is Radiation, after in loose cylindricality, the top capsule of conidial head
Spherical, top capsule is covered by the fusion overlapping layer of come into being stigma fused layer and bottle stalk, and bottle stalk produces conidia chain, conidium
Spherical or subsphaeroidal, wall is smooth.In view of above-mentioned morphological characteristic, Preliminary Identification bacterial strain CPCC400735 is aspergillus fungi.
1.2.2 molecular biology identification
Extracting the genomic DNA of bacterial strain CPCC400735, PCR expands 18S rRNA gene, and checks order.By gained sequence
(sequence 1 in sequence table) and sequence in GenBank data base are compared, and result shows bacterial strain CPCC400735 and aspergillosis
The similarity of (Aspergillus sp.) is 99%, therefore, in conjunction with before morphological characteristic, determine that bacterial strain CPCC400735 is
Aspergillus fungi.
Aspergillosis (Aspergillus sp.) CPCC400735 is preserved in Chinese microorganism strain on May 5th, 2016 and protects
Hiding administration committee's common micro-organisms center, preserving number is CGMCC No.12376.Hereinafter referred aspergillosis (Aspergillus
sp.)CPCC400735。
Embodiment 2, aspergillosis (Aspergillus sp.) CPCC400735 is utilized to prepare HIV-1 inhibitor
The present embodiment utilizes aspergillosis (Aspergillus sp.) CPCC400735 to be prepared for 5 kinds of HIV-1 inhibitor, wherein
A kind of HIV-1 inhibitor be that the methanolic extract of aspergillosis (Aspergillus sp.) CPCC400735 fermentation culture medium (uses first
Alcohol extracts the material being dissolved in methanol obtained from the culture of aspergillosis (Aspergillus sp.) CPCC400735), other 4
Planting HIV-1 inhibitor is the general formula compound shown in formula II
Formula II, its structural formula is specifically as shown in formula 1 to formula 4 (table 1).
The substituent group of general formula compound shown in table 1, formula II
The concrete preparation method of above-mentioned 5 kinds of HIV-1 inhibitor is as follows:
1, the culture of aspergillosis (Aspergillus sp.) CPCC400735 is prepared
Aspergillosis (Aspergillus sp.) CPCC400735 cultivates 5 days on 28 DEG C of PDA inclined-plane, and then picking mycelia block connects
Kind in equipped with 100ml seed culture medium (consisting of: glucose 2%, sucrose 1%, analysis for soybean powder 0.2%, peptone 1%, phosphoric acid
Hydrogen dipotassium 0.03%, Polyethylene Glycol 0.25%, sodium nitrate 0.3%, ammonium sulfate 0.3%, surplus is water, pH 6.0.121 DEG C of sterilizings
20 minutes, obtain seed culture medium) 500ml triangular flask in, 28-30 DEG C concussion cultivate 2 days as seed liquor.Then, will plant
Sub-liquid 10ml access 500ml triangular flask equipped with rice medium (500ml triangular flask equipped with 80g rice, add 100ml go from
Sub-water soaking, 121 DEG C of sterilizings obtain rice medium in 20 minutes) in, cultivate 40 days, obtain aspergillosis (Aspergillus for 28 DEG C
Sp.) CPCC400735 solid fermentation culture.
2, the methanolic extract of aspergillosis (Aspergillus sp.) CPCC400735 fermentation culture medium is prepared
Aspergillosis (Aspergillus sp.) the CPCC400735 solid fermentation culture Glass rod taking 2000g step 1 will
It blends, and adds 4L methanol, extracts 3 times at 28 DEG C, each 30min, merges methanol extract liquid, by Rotary Evaporators (temperature: 40
DEG C, revolution: 70 turns) rotary evaporation removes methanol in methanol extract liquid, and the material obtained is aspergillosis (Aspergillussp.)
The methanolic extract of CPCC400735 fermentation culture medium.
3, the compound shown in formula 1 to formula 5
3.1, the methanolic extract of aspergillosis (Aspergillus sp.) the CPCC400735 fermentation culture medium of step 2 is used
Water-dispersible, then it is extracted with ethyl acetate, collects ethyl acetate phase, revolve with Rotary Evaporators (temperature: 40 DEG C, revolution: 70 turns)
Turn evaporation remove ethyl acetate mutually in ethyl acetate, obtain extractum.Take after 50g extractum methanol dissolves and carry out silica gel column chromatography
Separate.Silica gel used in silica gel column chromatography is silica gel H, and the specification of silicagel column used is 6.5 × 20cm, and column volume is
663mL.Elution program used in silica gel column chromatography is first with 2 column volumes of chloroform eluting；Carry out again 13 column volumes as
Lower linear gradient elution: the mixed liquor that flowing used is made up of chloroform and methanol mutually, chlorine in linear gradient elution flowing mutually
Imitative and methanol volume ratio is linearly down to 7:1 by 20:1；Last two column volumes of again with methanol eluting.Start not from elution program
Being interrupted and collect the liquid eluted, often 200ml (200ml/ flows part) collected by pipe, collects 56 stream parts continuously, is designated as: Fr.1,
Fr.2, Fr.3, Fr.4 ..., Fr.56, TLC detection is instructed and is merged same stream part, finally obtains 12 and merges component: Fr.1-
3, Fr.4-12 (Fr.4 to Fr.12 these 9 stream part is merged and obtains, be the 0.9th to the 3.6th liquid that column volume elutes),
Fr.16 and Fr.17 these 2 stream part (is merged and obtains, be the 4.5th to the 5.1st cylinder by Fr.13-14, Fr.15, Fr.16-17
The long-pending liquid eluted), Fr.18-21, Fr.22-25, Fr.26-31, Fr.32-34, Fr.35-42, Fr.43-54, Fr.55-
56。
3.2, Fr.4-12 step 3.1 obtained carries out pressurized silica gel column chromatography.Silicon used in pressurized silica gel column chromatography
Glue is silica gel H, and the specification of silicagel column used is 4 × 13cm (a diameter of 4cm, a height of 13cm), and column volume is 163mL.Pressurization
Elution program used in silica gel column chromatography is by the following linear gradient elution of 6 column volumes: flowing used be mutually by
Petroleum ether and the mixed liquor of acetone composition, the petroleum ether in flowing mutually in linear gradient elution and the volume ratio of acetone are by 8:1
Linearly it is down to 5:1.Start uninterruptedly to collect the liquid eluted from elution program, often 20ml (20ml/ flows part) collected by pipe, continuously
Collect 43 stream parts, be designated as: Tube.1, Tube.2, Tube.3 ..., Tube.43.By Tube.27 to Tube.37 these 11
Stream part mixing, obtains the component (being the 3.2nd to the 4.5th liquid that column volume elutes) of entitled Tubes 27-37.
Tubes 27-37 is prepared type high performance liquid chromatography and separates (RP-C18 liquid phase preparative separation).This preparative
Filler used by high performance liquid chromatography separation is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 5 μm, this preparative
The a diameter of 8mm of chromatographic column, a height of 250mm used by high performance liquid chromatography separation.Flowing is mutually for the mixed liquor of first alcohol and water composition
Body, in flowing mutually, the volume ratio of first alcohol and water is 4: 1；Flow velocity is 4.0mL/min.Obtaining 6 preparation components, its title is respectively
tubes 27-37-P1(tR(retention time) is 6.0min), tubes 27-37-P2 (tRFor 23.9min), tubes27-37-P3
(tRFor 27.0min), tubes 27-37-P4 (tRFor 31.2min), tubes 27-37-P5 (tRFor 34-42min), tubes
27-37-P6(tRFor 44.6min).
Tubes 27-37-P2 is prepared type high performance liquid chromatography and separates (RP-C18 liquid phase preparative separation).This is prepared
Filler used by the separation of type high performance liquid chromatography is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 5 μm, and this is prepared
The a diameter of 8mm of chromatographic column, a height of 250mm used by the separation of type high performance liquid chromatography.Flowing is mutually for the mixing of first alcohol and water composition
Liquid, in flowing mutually, the volume ratio of first alcohol and water is 4: 1；Flow velocity is 4.5mL/min.Collect tR(retention time) is 23.9min
Eluting peak, with Rotary Evaporators (temperature: 40 DEG C, revolution: 70 turns) rotary evaporation remove first alcohol and water, obtain the formula 1 of 5.0mg
Shown compound.
Tubes 27-37-P6 is prepared type high performance liquid chromatography and separates (RP-C18 liquid phase preparative separation).This is prepared
Filler used by the separation of type high performance liquid chromatography is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 5 μm, and this is prepared
The a diameter of 8mm of chromatographic column, a height of 250mm used by the separation of type high performance liquid chromatography.Flowing is mutually for the mixing of first alcohol and water composition
Liquid, in flowing mutually, the volume ratio of first alcohol and water is 4: 1；Flow velocity is 5.0mL/min.Collect tR(retention time) is 37.7min
Eluting peak, with Rotary Evaporators (temperature: 40 DEG C, revolution: 70 turns) rotary evaporation remove first alcohol and water, obtain the formula of 14.7mg
Compound shown in 2.
3.3, Fr.16-17 step 3.1 obtained carries out pressurized silica gel column chromatography.Used by pressurized silica gel column chromatography
Silica gel is silica gel H, and the specification of silicagel column used is 4 × 12cm (a diameter of 4cm, a height of 12cm), and column volume is 151mL.Add
In pressure silica gel column chromatography, elution program used is the mixed of volume ratio 10:1 with the chloroform being made up of chloroform and methanol and methanol
Closing liquid and carry out eluting mutually as flowing, start from elution program uninterruptedly to collect the liquid eluted, often 10ml collected by pipe
(10ml/ flows part), collects 18 stream parts continuously, is designated as: Tube.1, Tube.2, Tube.3 ..., Tube.18.TLC instructs conjunction
And obtaining four merging components: tubes 1-3 (by Tube.1 to Tube.3 these 3 stream part mixing, obtains entitled tubes1-3
Component, be the liquid that elutes of the 0th to 0.2 column volume), Tube.4 to Tube.13 these 10 (is flowed part by tubes 4-13
Stream part mixing, obtains the component of entitled tubes 4-13, is that the 0.2nd column volume is to the 0.9th liquid that column volume elutes
Body), tubes 14-18.Carry out Tubes 4-13 again opening ODS column chromatography for separation.Used by this open ODS column chromatography for separation
Filler is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 50 μm, this open chromatography used by ODS column chromatography for separation
Column diameter is 4cm, a height of 16cm, and chromatographic column volume is 201mL.Elution program used by this opening ODS column chromatography for separation is two
Step eluting, first step eluting with the mixing liquid that volume ratio is 11:9 of the first alcohol and water being made up of first alcohol and water as flowing phase
4 column volumes of eluting, complete first step eluting；Second step eluting: after completing first step eluting, is carried out as flowing mutually with methanol
Eluting, starts to collect, from second step eluting, this 100ml liquid of 1-100ml that this second step eluting elutes and (uses methanol-eluted fractions
The 0th gone out is to the 0.5th liquid that column volume elutes), by its named Tubes 4-13-ODS-9.By Tubes 4-13-
ODS-9 with tubes 1-3 merging carries out preparative high performance liquid chromatography and separates (RP-C18 liquid phase preparative separation).This preparative is high
Filler used by effect liquid phase chromatogram separation is octadecylsilane chemically bonded silica filler, and the particle diameter of filler is 5 μm, and this preparative is high
The a diameter of 8mm of chromatographic column, a height of 250mm used by effect liquid phase chromatogram separation.The mixing liquid that flowing forms for first alcohol and water mutually,
In flowing mutually, the volume ratio of first alcohol and water is 3: 1；Flow velocity is 4.0mL/min.Obtain two preparation component P2 (tRFor 18.2min)
With P3 (tRFor 22.2min).Then, P2 with P3 is prepared type high performance liquid chromatography respectively under this identical liquid phase preparation condition
Separation is further purified.
The preparative high performance liquid chromatography of P2 separates and collects tR(retention time) is the eluting peak of 18.2min, steams with rotating
Send out instrument (temperature: 40 DEG C, revolution: 70 turns) rotary evaporation and remove first alcohol and water, obtain the compound shown in formula 4 of 52.0mg.
The preparative high performance liquid chromatography of P3 separates and collects tR(retention time) is the eluting peak of 22.2min, steams with rotating
Send out instrument (temperature: 40 DEG C, revolution: 70 turns) rotary evaporation and remove first alcohol and water, obtain the compound shown in formula 3 of 52.0mg.
The physicochemical property of the compound shown in 3.4 formula 1 to formulas 4
Compound shown in formula 1 to formula 4 is brown solid, is all soluble in methanol, ethanol, in DMSO equal solvent, and indissoluble
Yu Shui.Compound shown in formula 1 to formula 5 is at silica gel thin-layer chromatography chloroform-methanol-water (volume ratio 70:15:2) dicyandiamide solution
Expansion Rf value is 0.5-0.7, all obvious at 254nm and 365nm fluorescence developing, the aobvious brownish red of vanillin-sulfuric acid colour developing.
The structure elucidation of the compound shown in 3.5 formula 1 to formulas 4
HRESIMS (anion) quasi-molecular ions: the m/z 623.2900 [M-H] of the compound shown in formula 4-Point out its molecular formula
For C35H44O10.Comprehensive analysis1H NMR (Fig. 5),13C NMR (Fig. 6) and hsqc spectrum (Fig. 7), thus it is speculated that should be containing two in compound
Ketone carbonyl, an ester carbonyl group, 16 olefinic carbons (wherein 12 is quaternary carbon, and 4 is tertiary carbon, and has three carbon to connect oxygen atom) and 16
The carbon (wherein three connect oxygen atom) of individual sp2 hydridization.In hydrogen is composed visible features signal: δ 14.29 (1H, s), 13.07 (1H,
S), 11.97 (1H, brs), 10.69 (1H, brs), 6.79 (1H, s), 6.52 (1H, t), 4.90 (1H, t), 4.79 (1H, t),
2.80 (3H, s), 2.52 (1H, d), 2.11 (3H, s), 1.39 (3H, s), 1.24 (3H, s), 0.98 (3H, s), 0.92 (3H, s).
From features above hydrogen signal, it appeared that following coherent signal: δ 14.29 (13-OH) and 165.8 in HMBC spectrum (Fig. 8)
(C-13), 102.0 (C-3), 200.6 (C-2, weak) are relevant；δ 2.11 (H-14) and 107.2 (C-12), 162.9 (C-11),
165.8 (C-13) are correlated with；δ 13.07 (C-7-OH) and 163.3 (C-7), 118.0 (C-8), 106.1 (C-5), 202.8 (C-2,
Weak) relevant；δ 6.79 (8-H) and 106.1 (C-5), 112.8 (C-10), 26.1 (C-15), 163.3 (C-7), 202.8 (C-6,
Weak) relevant；δ 2.80 (H-15) is relevant with 112.8 (C-7), 118.0 (C-8), 149.0 (C-9)；δ 2.52 (H-16) and 81.2
(C-1), 200.6 (C-2), 202.8 (C-6), 115.4 (C-17), 139.6 (C-18) are relevant；δ 4.79 (H-17) and 15.6 (C-
35), 39.2 (C-19), 41.2 (C-16) are relevant；δ 1.24 (H-35) and 39.2 (C-19), 115.4 (C-17), 139.6 (C-18)
Relevant；δ 4.90 (H-17) is relevant with 15.3 (C-34), 25.9 (C-20), 38.0 (C-19)；δ 1.39 (H-34) and 38.0 (C-
23), 124.2 (C-21), 133.7 (C22) are relevant；δ 6.51 (H-25) and 23.9 (C-27), 24.4 (C-24), 38.0 (C-23),
168.6 (C-33) are correlated with；δ 3.00 (H-29) is relevant with 23.9 (C-27), 71.5 (C-30), 26.2 (C-31), 24.6 (C-32)；
δ 0.98 (H-31) is relevant with 24.6 (C-32), 71.5 (C-30), 77.3 (C-29)；δ 0.92 (H-32) and 26.2 (C-31),
71.5 (C-30), 77.3 (C-29) are relevant.?1H-1H COSY spectrum (Fig. 9) can find following coherent signal: δ 2.52 (H-16)
Relevant to 4.79 (H-17)；δ 1.60 (H-19) is relevant to 1.51 (H-20)；δ 1.51 (H-20) is relevant to 4.90 (H-21)；δ
1.93 (H-23) are relevant to 2.18 (H-24)；δ 2.18 (H-24) is relevant to 6.51 (H-25)；δ 3.00 (H-24) and 1.10 (H-
28b) relevant；δ 1.52H (H-28a) is relevant with 2.38 (H-27a), 2.09 (H-27b)；δ 1.10H (H-28b) and 2.38 (H-
27a), 2.09 (H-27b) are correlated with.According to information above, it may be determined that the planar structure of the compound shown in formula 4, hydrocarbon data
Ownership is shown in Table 2.In NOESY spectrum (Figure 10), coherent signal: δ 4.79 (H-17) is relevant to 1.60 (H-19)；δ 4.90 (H-21) with
1.93 (H-23) are correlated with；δ 2.18 (H-24) is relevant to 2.09 (H-27b), determines that in structure, three double bonds are trans (E) structure
Type.In structure, the configuration of two chiral carbon in C-1 and C-29 position is utilized respectively Rh2(OCOCF3)4Induction CD composes (Figure 11) and Mo
(Ac)4Induction CD spectrum (Figure 12) is identified.Rh at the compound shown in formula 42(OCOCF3)4In induction CD spectrum, aobvious at 347nm
It is shown as negative Cotton effect, according to document (Lorenzo DB, Gennaro P, Carmela P, et al.Determination
of Absolute Configuration of Acyclic 1,2-Diols with Mo2(OAc)4.1.Snatzke’s
Method Revisited [J] .J.Org.Chem.2001,66:4819-4825) the configuration decision rule reported, C-1 position exhausted
Should be R to configuration.Mo (Ac) at the compound shown in formula 44In induction CD spectrum, at 317nm, it is shown as negative Cotton effect
Should, according to document[32](Jadwiga Frelek,Wojciech J.Szczepek.[Rh2(OCOCF3)4]as an
auxiliary chromophore in chiroptical studies on steroidal alcohols[J]
.Tetrahedron:Asymmetry, 1999,10:1507-1520) the configuration decision rule reported, the absolute configuration of C-29 position
Should be R.Final compound shown in formula 4 be accredited as structure as shown in Figure 1.
HRESIMS (anion) quasi-molecular ions: the m/z 575.7247 [M-H] of the compound shown in formula 2-Point out its molecular formula
For C35H44O7.Compound shown in formula 21H and13C H NMR spectroscopy is the most as shown in Figs. 13 and 14.Compound shown in formula 2 and formula 1
The nuclear magnetic data of C-1～the C-22 structure fragment of shown compound is basically identical, and the nuclear-magnetism of C-23～C-33 structure fragment
Data have larger difference.(Figure 16): δ 5.02 (H-25) and 38.0 (C-23), 25.3 (C-24), 34.5 (C-27) in HMBC spectrum,
58.0 (C-33) are correlated with；δ 3.86 (H-33) is relevant with 125.2 (C-25), 139.0 (C-26), 34.5 (C-27)；δ5.02(H-
29) relevant with 34.5 (C-27), 17.5 (C-32)；δ 1.58 (H-31) is relevant with 124.4 (C-29), 134.0 (C-30), δ 1.50
(H-32) relevant with 124.4 (C-29), 134.0 (C-30).?1H-1H COSY to spectrum in (such as Figure 17): δ 5.02 (H-25) with
2.00 (H-24) are correlated with；δ 2.00 (H-24) is relevant to 1.82 (H-23)；δ 5.02 (H-29) is relevant to 1.98 (H-28)；δ1.98
(H-28) relevant to 1.96 (H-27), the structure of C-23 to C-33 fragment is can determine that according to information above.Comprehensive analysis1H-1H
COSY, HSQC (Figure 15) and HMBC spectrum, the planar structure of confirmation compound shown in formula 2, and hydrocarbon signal is belonged to (table
2).In NOESY spectrum (Figure 18), coherent signal: δ 4.77 (H-17) is relevant to 1.59 (H-19)；δ 4.84 (H-21) and 1.82 (H-
23) relevant；δ 2.00 (H-24) and 3.86 (H-33), prompting 17 (18) double bonds are trans (E) configuration, and 21 (22) double bonds are trans
(E) configuration, 25 (26) double bonds are cis (Z) configuration.Speculating according to biosynthesis pathway, the C-1's of the compound shown in formula 2 is exhausted
Should be identical with the compound shown in formula 4 to configuration, it is configured as R.Compound shown in formula 21H NMR (Figure 19),13C NMR
(Figure 22), hydrocarbon attribution data is shown in Table 2.Therefore, the structure being accredited as shown in Figure 2 of the compound shown in formula 2.
HRESIMS (anion) quasi-molecular ions: m/z 653.7920 [M-H] shown in formula 1-Its molecular formula is pointed out to be
C37H50O10.Compound shown in formula 11H and13C H NMR spectroscopy respectively as shown in Figure 19 and 20 and table 2, the change shown in contrast 1
Compound and the compound shown in formula 21H and13C NMR data finds, the two C-1 to C-22 structure fragment is identical, and C-23 is extremely
Differing greatly of C-33 structure fragment.In HMBC spectrum, (Figure 22): δ 0.95 (H-32) is relevant with 70.4 (C-30), 83.6 (C-29)；
δ 1.01 (H-31) is relevant with 70.4 (C-30), 83.6 (C-29)；δ 3.24 (H-36) and δ 3.23 (H-37) respectively with 106.0 (C-
33 are correlated with)；δ 4.12 (H-33) is relevant with 77.2 (C-25), 43.0 (C-26), 23.0 (C-27)；δ 2.76 (H-29) and 23.0
(C-27) relevant；δ 3.02 (H-25) is relevant to 34.6 (C-23).?1H-1H COSY spectrum in (Figure 23): δ 4.12 (H-33) with
1.37 (H-26) are correlated with, and δ 1.37 (H-26) is relevant with 3.02 (H-25) and 1.75 (H-27a), 1.22 (H-27b) respectively；3.02
(H-25) relevant to 1.25 (H-24b)；1.70 (H-24a) are relevant to 1.98 (H-23a).Degree of unsaturation according to compound and point
Minor speculates, there is a circulus, δ 2.76 (H-29) and 77.2 in composing according to HMBC in C-23 to C-33 structure fragment
(C-27) it is correlated with and δ 3.02 (H-29) and 83.6 (C-29) relevant information, determines in this structure fragment it is that C-25 and C-29 position leads to
Cross single oxygen bonded formation ether ring.The structure of C-23 to C-33 fragment is may determine that according to information above.Comprehensive analysis1H-1H
COSY, HSQC (Figure 21) and HMBC spectrum, determine the planar structure of compound shown in formula 1, and hydrocarbon signal belonged to (table
2).In NOESY spectrum (Figure 24), coherent signal: δ 4.77 (H-17) is relevant to 1.60 (H-19)；δ 4.86 (H-21) and 1.88 (H-
23b) relevant, prompting 17 (18) double bonds and 21 (22) double bonds are trans (E) configuration.Speculate according to biosynthesis pathway, shown in formula 1
The absolute configuration of C-1 of compound should be identical with the compound shown in formula 4, it is configured as R.C-25 in structure, C-26 and
C-29 tri-is chiral carbon, during its absolute structure is calculated by ECD.Coherent signal in composing according to NOESY, determines C-23
The relative configuration (Figure 25) of the hexa-atomic ether ring to C-33 structure fragment.Finally, the compound shown in formula 1 be accredited as figure
Structure shown in 3.
HRESIMS (anion) quasi-molecular ions: the m/z 609.3094 [M-H] of the compound shown in formula 3-Point out its molecular formula
For C35H46O9.Compound shown in formula 31H and13C H NMR spectroscopy is the most as shown in figures 26 and 27.With the compound phase shown in formula 4
Ratio, relatively it has lacked an oxygen atom to the compound shown in formula 3, many 2 hydrogen atoms.The two carbon modal data of contrast, shown in formula 3
Compound is fewer than the compound shown in a formula 4 ester carbonyl group carbon atom signal, the saturated carbon atom of many even oxygen atoms
Signal δ 58.2, and other carbon modal data are basically identical, the C-33 position of the compound shown in prompting type 3 is probably a methylol.
Comprehensive analysis1H-1H COSY (Figure 30), HSQC (Figure 28) and HMBC compose (Figure 29), identify the planar junction of compound shown in formula 3
Structure, and hydrocarbon signal is belonged to (table 2).Coherent signal: δ 4.78 (H-17) and 1.59 (H-19) in NOESY (Figure 31) spectrum
Relevant；δ 4.85 (H-21) is relevant to 1.83 (H-23)；δ 2.00 (H-24) and 3.85 (H-33), prompting 17 (18) double bonds are trans
(E) configuration, 21 (22) double bonds are trans (E) configuration, and 25 (26) double bonds are cis (Z) configuration.Speculate according to biosynthesis pathway,
The absolute configuration of C-1 with C-29 of the compound shown in formula 1 should be identical with the compound shown in formula 4, and its configuration is R.Therefore,
Compound shown in formula 3 is accredited as structure as shown in Figure 4.
The nuclear magnetic data of the compound shown in table 2, formula 1 to formula 4 (1H NMR 600MHz；13C NMR 150MHz；DMSO-
d6)
4, the inhibitory activity to HIV-1
(1) cell is cultivated and transfection: 293T cell is incubated in the DMEM culture medium containing 10%FBS.6 orifice plate every hole inoculations
Cell number is 4 × 105Individual, transfect after cultivating 24h, transfection reagent Lipofectamine 2000, carry out turning according to operation instruction
Dye；SupT1 cell (SUP-T1 cell, human T lymphoma cell) is incubated in RPMI 1640 culture medium containing 10%FBS, is placed in
The 5%CO of 37 DEG C2Quiescent culture in incubator.
(2) preparation of HIV-1 pseudovirus: be 1.5 × 10 by cell concentration5The amount of individual/ml by 293T cell before transfection
Within one day, it is inoculated in 6 orifice plates, in 2mL culture medium, cultivates 24h transfect.Plasmid consumption is: transfection pNL4-in every hole in six orifice plates
3Luc (R-E-) 300ng and pHIT/G plasmid 210ng, transfection reagent is Lipofectamine2000, by transfection reagent illustrate into
Row transfection.Collect supernatant after 48 hours, cross the filter membrane of 0.45 μm, collect virus liquid and i.e. obtain VSV-G coated HIV-1 cape horn fever
Poison.
(3) mensuration of viral infection: 96 orifice plate every hole inoculations 5 × 105The SupT1 cell 200 μ L of individual/ml.Take 10 μ l
Gained virus liquid infects SupT1 cell.After cultivating 48h, take out 50 μ l cell suspension, add equal-volume 2 × Luciferase
Cell Culture Lysis cell lysis.Test kit Luciferiase Assay System is used to measure the fluorescence of lysate
Element enzyme (Luciferase) activity.Size according to reporter gene Luciferase activity draws the amount of infectious virus.
(4) Anti-HIV-1 Active measures: SupT1 cell concentration is 5 × 105Individual/ml, directly paving 96 orifice plates, every hole 200 μ L,
It is simultaneously introduced the HIV-1 pseudovirus (virus is 1:20 with the volume ratio of cell) of above-mentioned preparation.(sample is dissolved in add sample
In DMSO), each sample sets 3 repetitions, arranges blank and negative control (DMSO) simultaneously, hatches 48h for 37 DEG C.Take 50 μ L
Cell suspension, adds 2 × Luciferase Cell Culture Lysis lysate 50 μ L, after 37 DEG C hatch half an hour, takes
Wherein 6 μ L cell pyrolysis liquids add 40 μ L luciferase substrate, utilize Centro XS3LB 960 microplate reader to measure luciferase
Value (RLUs), calculates the sample suppression ratio to HIV.
HIV suppression ratio computing formula is as follows:
Wherein, sample is the methanol of aspergillosis (Aspergillus sp.) CPCC400735 fermentation culture medium prepared by step 2
Any one in these 6 kinds of HIV-1 inhibitor of the compound shown in formula 1 to formula 5 of extract and step 3 preparation.By sample pair
The suppression ratio of HIV-1, calculates the IC of the compound shown in formula 1 to formula 550(refer to that Inhibit the replication of HIV-1 reduces by the medicine of 50%
Concentration).
(5) drug toxicity detection utilizes Cell Counting Kit-8 (CCK-8 test kit) to detect drug toxicity: SupT1
Cell concentration is 5 × 105Individual/ml, directly paving 96 orifice plates, every hole 100 μ L.Add sample (sample is dissolved in DMSO), each
Sample sets 3 repetitions, arranges blank and negative control (DMSO) simultaneously, hatches 48h for 37 DEG C.Taking out 96 orifice plates, every hole adds
Enter 10 μ L CCK-8, after 37 DEG C are continued to hatch 1 hour, use the multi-functional microplate reader of Enspire 2300 to detect each hole and exist
Absorbance value (OD) at 450nm wavelength, calculates the cell survival rate of sample well.Cell survival rate computing formula:
Wherein, any one during sample is these 4 kinds of HIV-1 inhibitor of the compound shown in formula 1 to formula 4 of step 3 preparation.
By the cell survival rate of sample well, calculate the CC of the compound shown in formula 1 to formula 450(refer to cause 50% cell death
Drug level).
Result shows that the methanol of aspergillosis (Aspergillus sp.) CPCC400735 fermentation culture medium prepared by step 2 carries
Taking the thing suppression ratio when concentration is 4mg/mL to HIV-1 is 99.9%.Compound shown in formula 1 to formula 4 is 100 μMs in concentration
Time the suppression ratio of HIV-1 is respectively 99.84%, 99.99%, 98.60% and 96.82%, the compound shown in formula 1 to formula 4
IC50It is respectively 2.4 μMs, 4.5 μMs, 22.1 μMs and 32.6 μMs, the CC of the compound shown in formula 1 to formula 350It is all higher than 100 μMs,
The CC of the compound shown in formula 450It is 78.6 μMs (tables 3).Aspergillosis (Aspergillus sp.) prepared by step 2 is described
Compound shown in the methanolic extract of CPCC400735 fermentation culture medium and formula 1 to formula 4 has stronger suppression and lives HIV-1
Property, and toxicity is less.Wherein, the compound shown in formula 1 is the strongest to the inhibitory activity of HIV-1, the compound pair shown in formula 2
The inhibitory activity of HIV-1 is weaker than the compound shown in formula 1 but is better than the compound shown in formula 3 and formula 4, the compound shown in formula 3
The inhibitory activity of HIV-1 is better than the compound shown in formula 4.
The Anti-HIV-1 Active of the compound shown in table 3, formula 1 to formula 4
| Compound number | Average inhibition % (100 μMs, n=3) | IC<sub>50</sub>(μM) | CC<sub>50</sub>(μM) |
| Formula 1 | 99.84±0.12 | 2.4 | >100 |
| Formula 2 | 99.99±0.01 | 4.5 | >100 |
| Formula 3 | 98.60±0.53 | 22.1 | >100 |
| Formula 4 | 96.82±0.11 | 32.6 | 78.6 |
Claims (10)
1. aspergillosis, its bacterial strain number is CPCC400735, and it is in China Committee for Culture Collection of Microorganisms's common micro-organisms
The numbered CGMCC No.12376 that registers on the books of the heart.
2.U1) or U2) application:
U1) application in preparing hiv inhibitor of the aspergillosis described in claim 1；
U2) aspergillosis described in claim 1 is treated or/and prevent answering in acquired immune deficiency syndrome (AIDS) medicine in preparation
With.
3.P1) or P2) product:
P1) hiv inhibitor prepared by the aspergillosis described in claim 1；
P2) treatment prepared by the aspergillosis described in claim 1 is or/and prevent acquired immune deficiency syndrome (AIDS) medicine.
4. the culture of aspergillosis described in claim 1, is to be cultivated in microbiological culture media by the aspergillosis described in claim 1
Material in the culture vessel arrived.
Culture the most according to claim 4, it is characterised in that: described microbiological culture media is aspergillosis culture medium.
6.P4) or P5) product:
P4) hiv inhibitor, it is characterised in that: the active component of described hiv inhibitor is from described in claim 4 or 5 with methanol
Culture in extract the material being dissolved in methanol that obtains；
P5) treatment is or/and prevent acquired immune deficiency syndrome (AIDS) medicine, it is characterised in that: described treatment is or/and prevention obtains
The active component of property immunodeficiency syndrome medicine is to extract from the culture described in claim 4 or 5 with methanol to obtain
It is dissolved in the material of methanol.
7.M1) or M2) method:
M1) method preparing hiv inhibitor, is dissolved in including extracting from the culture described in claim 4 or 5 with methanol
The material of methanol, using described material as the active component of hiv inhibitor；
M2) preparation treatment or/and prevention acquired immune deficiency syndrome (AIDS) medicine method, including with methanol from claim 4
Or extraction obtains being dissolved in the material of methanol in the culture described in 5, using described material as treatment or/and prevent acquired immunity
The active component of deficit syndrome medicine.
8.M4) or M5) method:
M4) method preparing hiv inhibitor, including extracting obtain molten from the culture described in claim 4 or 5 with methanol
In the material of methanol, then extract from described material and obtain the compound shown in formula II, using the compound shown in formula II as HIV
The active component of inhibitor；
Described R is
M5) preparation treatment or/and prevention acquired immune deficiency syndrome (AIDS) medicine method, including with methanol from claim 4
Or in the culture described in 5, extraction obtains being dissolved in the material of methanol, then extraction obtains the chemical combination shown in formula II from described material
Thing, using the compound shown in formula II as treatment or/and prevent the active component of acquired immune deficiency syndrome (AIDS) medicine；
Described R is
9.1) to 10) in any one:
1) compound shown in formula II in method described in claim 8 or its pharmaceutically acceptable salt；
2) compound shown in formula II in method described in claim 8 or its pharmaceutically acceptable salt are in preparation HIV suppression
Application in agent；
3) compound shown in formula II in method described in claim 8 or its pharmaceutically acceptable salt preparation treatment or/
With the application in prevention acquired immune deficiency syndrome (AIDS) medicine；
4) hiv inhibitor, its active component be the compound shown in the formula II in method described in claim 8 or its pharmaceutically may be used
The salt accepted；
5) treatment is or/and prevention acquired immune deficiency syndrome (AIDS) medicine, and its active component is in method described in claim 8
Compound shown in formula II or its pharmaceutically acceptable salt；
6) method preparing the compound shown in the formula II in method described in claim 8, including with methanol from claim 4 or
Culture described in 5 extracts the material being dissolved in methanol obtained, then extraction obtains the chemical combination shown in formula II from described material
Thing；
7) treatment is or/and the method for prevention acquired immune deficiency syndrome (AIDS), uses with methanol from right including to receptor
Require the culture described in 4 or 5 extracts to obtain being dissolved in the material of methanol, carry out treating or/and prevent acquired immunodeficiency
Syndrome；
8) method suppressing HIV animal, uses with methanol from the cultivation described in claim 4 or 5 including to receptor
Thing extracts the material being dissolved in methanol obtained to suppress HIV animal；
9) treatment is or/and the method for prevention acquired immune deficiency syndrome (AIDS), uses described in claim 8 including to receptor
The compound shown in formula II or its pharmaceutically acceptable salt in method carry out treating or/and prevent acquired immunodeficiency to combine
Simulator sickness；
10) method suppressing HIV animal, uses shown in the formula II in method described in claim 8 including to receptor
Compound or its pharmaceutically acceptable salt to suppress HIV animal.
Product described in application the most according to claim 2, claim 3 or 6, the cultivation described in claim 4 or 5
Method described in thing, claim 7 or 8 or 1 described in claim 9) to 10) in any one, it is characterised in that: described HIV
For the HIV of HIV-1 type, described acquired immune deficiency syndrome (AIDS) is caused by HIV-1.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910609383.5A CN110305798B (en) | 2016-06-15 | 2016-06-15 | Application of aspergillus, compound prepared from aspergillus and application of compound |
| CN201610424586.3A CN106085868B (en) | 2016-06-15 | 2016-06-15 | One plant of Aspergillus and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610424586.3A CN106085868B (en) | 2016-06-15 | 2016-06-15 | One plant of Aspergillus and its application |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910609383.5A Division CN110305798B (en) | 2016-06-15 | 2016-06-15 | Application of aspergillus, compound prepared from aspergillus and application of compound |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106085868A true CN106085868A (en) | 2016-11-09 |
| CN106085868B CN106085868B (en) | 2019-07-12 |
Family
ID=57846336
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610424586.3A Active CN106085868B (en) | 2016-06-15 | 2016-06-15 | One plant of Aspergillus and its application |
| CN201910609383.5A Active CN110305798B (en) | 2016-06-15 | 2016-06-15 | Application of aspergillus, compound prepared from aspergillus and application of compound |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910609383.5A Active CN110305798B (en) | 2016-06-15 | 2016-06-15 | Application of aspergillus, compound prepared from aspergillus and application of compound |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN106085868B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108420815A (en) * | 2017-02-14 | 2018-08-21 | 中国医学科学院医药生物技术研究所 | Application of the polyketone in inhibiting influenza virus |
| CN108822119A (en) * | 2018-06-25 | 2018-11-16 | 中国医学科学院医药生物技术研究所 | Red alcohol ketone compounds, preparation method and its pharmacy application with autophagy Activation Activity |
| CN110251656A (en) * | 2019-07-04 | 2019-09-20 | 自然资源部第三海洋研究所 | The drug of the latent activation of the preparation method and applications and HIV of polypeptide compound |
| CN113651683A (en) * | 2021-08-06 | 2021-11-16 | 中国医学科学院医药生物技术研究所 | Perylene ketone compound and application thereof |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106085868B (en) * | 2016-06-15 | 2019-07-12 | 中国医学科学院医药生物技术研究所 | One plant of Aspergillus and its application |
| CN114306394A (en) * | 2020-09-30 | 2022-04-12 | 杭州远大生物制药有限公司 | Microbial preparation and preparation method thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20150048759A (en) * | 2012-09-07 | 2015-05-07 | 노파르티스 아게 | Indole carboxamide derivatives and uses thereof |
| CN106085868B (en) * | 2016-06-15 | 2019-07-12 | 中国医学科学院医药生物技术研究所 | One plant of Aspergillus and its application |
-
2016
- 2016-06-15 CN CN201610424586.3A patent/CN106085868B/en active Active
- 2016-06-15 CN CN201910609383.5A patent/CN110305798B/en active Active
Non-Patent Citations (3)
| Title |
|---|
| XU PANG等: "Metabolites from the plant endophytic fungus Aspergillus sp. CPCC 400735 and their anti-HIV activities", 《JOURNAL OF NATURAL PRODUCTS》 * |
| 施琦渊等: "植物内生真菌来源的抗菌活性物质研究进展", 《中国药学杂志》 * |
| 杨志钧等: "《第四届全国微生物资源学术暨国家微生物资源平台运行服务研讨会论文集》", 26 November 2012 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108420815A (en) * | 2017-02-14 | 2018-08-21 | 中国医学科学院医药生物技术研究所 | Application of the polyketone in inhibiting influenza virus |
| CN108420815B (en) * | 2017-02-14 | 2019-12-31 | 中国医学科学院医药生物技术研究所 | Application of polyketone in inhibiting influenza virus |
| CN108822119A (en) * | 2018-06-25 | 2018-11-16 | 中国医学科学院医药生物技术研究所 | Red alcohol ketone compounds, preparation method and its pharmacy application with autophagy Activation Activity |
| CN108822119B (en) * | 2018-06-25 | 2020-08-28 | 中国医学科学院医药生物技术研究所 | Erythronolide ketone compound with autophagy activation activity, preparation method and pharmaceutical application thereof |
| CN110251656A (en) * | 2019-07-04 | 2019-09-20 | 自然资源部第三海洋研究所 | The drug of the latent activation of the preparation method and applications and HIV of polypeptide compound |
| CN110251656B (en) * | 2019-07-04 | 2022-06-10 | 自然资源部第三海洋研究所 | Preparation method of polypeptide compound, application of polypeptide compound and HIV latent activation medicine |
| CN113651683A (en) * | 2021-08-06 | 2021-11-16 | 中国医学科学院医药生物技术研究所 | Perylene ketone compound and application thereof |
| WO2023011668A1 (en) * | 2021-08-06 | 2023-02-09 | 中国医学科学院医药生物技术研究所 | Perinaphthenone compound and use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110305798A (en) | 2019-10-08 |
| CN110305798B (en) | 2021-11-19 |
| CN106085868B (en) | 2019-07-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106085868B (en) | One plant of Aspergillus and its application | |
| CN102168022B (en) | Endophytic fungus Penicillium ateckii from plant Chinese Umbrellaleaf rhizome and application thereof | |
| CN107298671A (en) | Come from the secalonic acid H of penicillium oxalicum and prepare the application of anti-human colon cancer drug | |
| CN102559828B (en) | Method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms | |
| CN103910746B (en) | The Berkeleyones compound in one class marine fungi source and its preparation method and application | |
| CN107298672A (en) | The secalonic acid I for coming from penicillium oxalicum is preparing the application of anti-human colon cancer drug | |
| CN106086147A (en) | Fungal elicitor is utilized to induce the method extracting betulic acid from Inonqqus obliquus | |
| CN106047713A (en) | Talaromyces pinophilum strain Li-93 and application thereof | |
| CN107298670A (en) | Come from penicillium oxalicum secalonic acid H and prepare anti-human oral cavity epidermoid carcinoma medicinal application | |
| CN105018352A (en) | Fungal strain capable of producing kojic acid and preparation method of fungal strain | |
| CN101234951A (en) | Biphenyls compound and its preparation method and application | |
| CN104087645A (en) | Solid fermentation ginseng flower and preparation method thereof | |
| CN106119333A (en) | A kind of method utilizing fatty acid-induced to extract betulic acid from Inonqqus obliquus | |
| CN103911407B (en) | The preparation method of the Azaphilone class dimer compound in a kind of marine fungi source and application | |
| CN107298669A (en) | Come from the secalonic acid I of penicillium oxalicum and anti-human oral cavity epidermoid carcinoma medicinal application | |
| CN101487022A (en) | Preparation of fermentation liquor for inhibiting liver cancer cell growth | |
| CN107513072B (en) | Polyketone is inhibiting the application in HIV | |
| CN105420119A (en) | Ginseng endophytic fungus and application thereof | |
| CN109234175A (en) | The Fusarium oxysporum DCLZJ-4 bacterial strain and application thereof of one plant of production chonglou saponin | |
| CN101603011A (en) | The method and the special strain therefore thereof that prepare the two glucosides of epipodophyllotoxin | |
| CN107954839A (en) | A kind of antiinflammatory active compound peniroquesine A and its preparation method and application | |
| CN102584615A (en) | Alkaloid compound as well as preparation method and application thereof | |
| CN110343618B (en) | Helminthosporol type sesquiterpenoids, and preparation method and application thereof | |
| CN107034253A (en) | A kind of method that use Yunnan rough gentian endogenetic fungus carries out bioconversion | |
| CN105316238B (en) | A method of the culture screening taxol-producing fungi from Chinese yew |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |