[go: up one dir, main page]

CN107510694B - Compound for activating latent infected AIDS virus and application thereof in AIDS treatment - Google Patents

Compound for activating latent infected AIDS virus and application thereof in AIDS treatment Download PDF

Info

Publication number
CN107510694B
CN107510694B CN201610428916.6A CN201610428916A CN107510694B CN 107510694 B CN107510694 B CN 107510694B CN 201610428916 A CN201610428916 A CN 201610428916A CN 107510694 B CN107510694 B CN 107510694B
Authority
CN
China
Prior art keywords
hiv
compound
active ingredient
formula
cells
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.)
Active
Application number
CN201610428916.6A
Other languages
Chinese (zh)
Other versions
CN107510694A (en
Inventor
王建华
孙丽
蒋金凤
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Institute Of Immunology And Infection Chinese Academy Of Sciences
Original Assignee
Institut Pasteur of Shanghai of CAS
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Institut Pasteur of Shanghai of CAS filed Critical Institut Pasteur of Shanghai of CAS
Priority to CN201610428916.6A priority Critical patent/CN107510694B/en
Publication of CN107510694A publication Critical patent/CN107510694A/en
Application granted granted Critical
Publication of CN107510694B publication Critical patent/CN107510694B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/407—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with other heterocyclic ring systems, e.g. ketorolac, physostigmine
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明提供了一种艾滋病病毒潜伏激活剂其应用。具体地，提供了本发明式I化合物或其药学上可接受的盐，所述化合物具有优异的激活潜伏感染的艾滋病病毒的效果。本发明还提供了含有式I化合物的药物组合物以及在激活潜伏感染的艾滋病病毒方面的应用。

Figure DDA0001020081440000011
The invention provides an HIV latent activator and its application. Specifically, there is provided a compound of formula I of the present invention or a pharmaceutically acceptable salt thereof, which has an excellent effect of activating latently infected HIV. The present invention also provides a pharmaceutical composition containing the compound of formula I and its use in activating latently infected HIV.
Figure DDA0001020081440000011

Description

Compound for activating latent infected AIDS virus and application thereof in AIDS treatment
Technical Field
The invention relates to the field of medicinal chemistry, in particular to a compound for activating latent infected AIDS virus and application thereof in AIDS treatment.
Background
After the first AIDS cases were discovered in the United states in 1981, AIDS spread rapidly worldwide. According to the statistics of the United nations AIDS planning administration, 3900 thousands of people die of AIDS all over the world. By the end of 2014, about 3690 million people worldwide carry HIV. Among total infected people, the proportion of sub-saharan in south Africa is the highest, about 71%; the second is asian and pacific regions, about 14%.
Currently, there is still a lack of effective drugs for the radical cure of HIV infection worldwide. The current stage of treatment targets are: the virus load is reduced to the maximum extent and durably; obtaining immune function reconstruction and maintaining immune function; the quality of life is improved; reducing HIV-associated morbidity and mortality.
Highly effective antiretroviral therapy (HAART) can effectively control the replication of human immunodeficiency virus type 1 (HIV-1) in AIDS patients, but cannot eradicate latent infection viruses, which becomes one of the main difficulties of current AIDS treatment. Some immunization or gene therapy approaches are proposed against latent HIV-1 infection, and some strategies are in clinical trials, including "activation-killing" mode, in which an epigenetic modification inhibitor or some signal pathway activator is used to activate latent HIV-1, and then the activated virus is killed in combination with HAART, so as to achieve the purpose of eliminating the latent virus. Various HIV-1 latent activator compounds such as the epigenetic modification inhibitor SAHA (also known as Vorinostat) show good effects at the cellular level, but clinical trials have not achieved the expected significant effects. There is an urgent need in the art for the extensive screening of highly potent, low-toxicity HIV-1 latently activating compounds.
Disclosure of Invention
The invention aims to provide a compound for activating latent infected Human Immunodeficiency Virus (HIV) and application thereof in AIDS treatment.
In a first aspect of the invention, there is provided the use of a compound of formula I, or a derivative or pharmaceutically acceptable salt thereof, for the preparation of a pharmaceutical composition or formulation for the activation of a latently infected HIV,
Figure BDA0001020081420000021
in another preferred embodiment, the compound of formula I is compound FICZ (6-formylindolo [3,2-b ] carbazole).
In another preferred embodiment, said activating a latently infected HIV comprises activating HIV replication.
In another preferred embodiment, the HIV comprises HIV-1 virus.
In another preferred embodiment, the latent infection comprises a resting infection.
In another preferred embodiment, the latent infection comprises a latent infection in an immune cell.
In another preferred embodiment, the immune cell is selected from the group consisting of: t cells, and HK cells.
In another preferred embodiment, the T cells comprise CD4+T cell line and CD4 isolated from human+T cells
In another preferred embodiment, the T cells comprise CD4+T cells.
In another preferred embodiment, said compound of formula I has one or more properties selected from the group consisting of:
(i) at a concentration of 50 μ M, the cell viability is greater than 60%, preferably greater than 80%;
(ii) no non-specific immune activation.
In another preferred embodiment, the pharmaceutical composition further comprises other active ingredients for treating AIDS.
In another preferred embodiment, the pharmaceutical composition further comprises an antiretroviral drug.
In another preferred embodiment, the antiretroviral drug is selected from the group consisting of: an HIV reverse transcriptase inhibitor, an HIV integrase inhibitor, an HIV proteolytic enzyme inhibitor, an HIV-cell membrane fusion inhibitor, an HIV-co-acceptor binding inhibitor, or a combination thereof.
In another preferred embodiment, the pharmaceutical composition or preparation is also used for treating AIDS.
In another preferred embodiment, the pharmaceutical composition comprises 0.001-99 wt%, preferably 0.1-90 wt%, more preferably 1-80 wt% of the compound of formula I or its pharmaceutically acceptable salt, based on the total weight of the composition.
In another preferred embodiment, the dosage form of the pharmaceutical composition is an oral dosage form or an injection.
In another preferred example, the oral dosage form comprises tablets, capsules, films, granules and the like, and also comprises sustained-release or non-sustained-release dosage forms.
In a second aspect of the invention, there is provided a pharmaceutical composition comprising: an active ingredient (a) a compound of formula I, or a derivative thereof, or a pharmaceutically acceptable salt thereof;
Figure BDA0001020081420000031
an active ingredient (b) an antiretroviral drug;
and (c) a pharmaceutically acceptable carrier.
In another preferred embodiment, the ratio (mg: mg) of the active ingredient (a) to the active ingredient (b) is 1:100 to 100:1, preferably 1:20 to 20: 1.
In another preferred embodiment, the total content of the active ingredient (a) and the active ingredient (b) is 1 to 99 wt%, and more preferably 5 to 90 wt% of the composition.
In another preferred embodiment, the pharmaceutical composition is used for treating AIDS.
In a third aspect of the invention, there is provided a kit comprising:
(i) a first container, and an active ingredient (a) a compound of formula I, or a derivative or pharmaceutically acceptable salt thereof, or a medicament containing the active ingredient (a) contained in the first container;
(ii) a second container, and an active ingredient (b) an antiretroviral drug, or a drug containing an active ingredient (b), contained in the second container; and
(iii) instructions for the combined administration of active ingredient (a) and active ingredient (b) for the treatment of AIDS are described.
In another preferred embodiment, the medicament in the first container and the second container is a single preparation containing the active ingredient (a) and a single preparation containing the active ingredient (b).
In another preferred embodiment, the dosage form of the drug is oral dosage form or injection.
In a fourth aspect of the present invention, there is provided a method for activating a latently infected HIV in vitro, comprising the steps of:
(a) contacting a cell latently infected with an HIV with a compound of formula I, or a derivative or pharmaceutically acceptable salt thereof, according to the first aspect of the invention, thereby activating the latently infected HIV.
In another preferred embodiment, the method is non-therapeutic.
In another preferred embodiment, in step (a), the compound of formula I, or a derivative thereof, or a pharmaceutically acceptable salt thereof, is added to a cell culture system, such that it is contacted with cells latently infected with hiv.
In another preferred embodiment, the cell is an immune cell.
In another preferred embodiment, the cell is a human or mammalian cell.
In a fifth aspect of the present invention, there is provided a method of activating a latently infected aids virus, said method comprising:
(i) administering to a subject in need thereof a compound of formula I as described in the first aspect of the invention, or a derivative thereof, or a pharmaceutically acceptable salt thereof.
In another preferred embodiment, the subject includes humans and non-human mammals (e.g., rodents and primates).
It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.
Drawings
FIG. 1 shows the structural formula of compound FICZ.
FIG. 2 shows FICZ activating HIV-1 latently infected Jurkat CD4+T cells.
FIG. 2A shows flow cytometry quantification of HIV-1 latent infection by either FICZ or SAHA compounds at various concentrations in Jurkat CD4+T cell activation effect.
Fig. 2B shows a concentration-dependent curve and the calculated EC50 plotted from the results of fig. 2A.
Figure 3 shows that FICZ has lower cytotoxicity. FICZ and HIV-1 latently infected Jurkat CD4+T cells were co-cultured for 48 hours and cell survival was examined using the MTT method.
FIG. 4 shows FICZ activates HIV-1 CD4 in a resting infection+Replication in T cells. Isolation of peripheral blood statically infected CD4 from HIV-1 infected persons+T cells. Treatment of statically infected CD4 with Compound FICZ+After 48 hours on T cells, RNA was collected and HIV-1 replication was detected by quantifying the level of HIV-1gag mRNA. The figure shows 4 patients CD4+T cell results.
Figure 5 shows FICZ cytotoxicity. FICZ treatment of HIV-1 infected purified statically infected CD4+T cells, cells were collected after 48 hours, stained with Trypan Blue (Trypan Blue), and cell survival was counted.
FIG. 6 shows FICZ vs. primary CD4+Immune activation of T cells. Healthy human PBMC cells were treated with FICZ at high concentration for 48 hours, and positive cell rates were calculated by detecting cell surface CD25 and CD69 molecular expression using flow cytometry.
Detailed Description
The inventor of the invention has studied extensively and deeply, discovered a kind of compound shown in the structural formula I can activate latent infection AIDS virus for the first time unexpectedly. Experiments show that the compound of the formula I has excellent activity of activating latent infected AIDS virus, low cytotoxicity and no non-specific activation effect on immune cells. The FICZ can also be combined with antiretroviral drugs for treatment, is used for eliminating latent HIV-1, and has clinical application value. On the basis of this, the present invention has been completed.
Specifically, the invention detects the activation effect of FICZ on latent HIV-1, and the experiment shows that FICZ can activate Jurkat CD4 of HIV-1 latent infection+T cells, half Effective Concentration (EC)50) At 47nM, the activation activity was 15 times that of SAHA; the FICZ cytotoxicity is detected by using an MTT method, and the cells still maintain the cell survival rate of up to 82% when the concentration of the FICZ is up to 50 mu M, which shows that the FICZ has lower cytotoxicity; treatment of HIV-1 with 4 antiretroviral drugsResting CD4 isolated from infected persons+T cells, FICZ was found to have 2.6-60 fold activity to activate HIV-1 replication and to exhibit lower cytotoxicity; FICZ unstimulated CD69 and CD25 molecules in primary CD4+Expression on T cells, FICZ, showed no non-specific activation on immune cells. Taken together, the results show that the compound FICZ can efficiently activate HIV-1 latent infection, and shows low cytotoxicity and no nonspecific immune activation. The FICZ can be combined with antiviral treatment, is used for eliminating latent HIV-1, and has clinical application value.
Term(s) for
Compounds of the invention
As used herein, the term "compound of the present invention", "compound FICZ", or "FICZ" refers to a compound of formula I, or a racemate, a enantiomer, or a pharmaceutically acceptable salt thereof. It is to be understood that the term also includes mixtures of the above components.
Figure BDA0001020081420000061
FICZ (6-Formyllindolo [3,2-b ] carbazole, i.e. 6-Formylindolo [3,2-b ] carbazole), is a high affinity aromatic receptor AHR (aryl hydrocarbon receptor) ligand derived from tryptophan. The compound has a skeleton structure of indole [3,2-b ] carbazole, and aldehyde group is introduced into 6 position.
In the present invention, pharmaceutically acceptable salts of the compounds of formula I are also included. The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts include inorganic and organic salts. One preferred class of salts is that formed by reacting a compound of the present invention with an acid. Suitable acids for forming the salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, phenylmethanesulfonic acid, benzenesulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid.
Antiretroviral agents
Antiretroviral drugs include HIV reverse transcriptase inhibitors, HIV integrase inhibitors, HIV proteolytic enzyme inhibitors, HIV-to-cell membrane fusion inhibitors, HIV-to-co-receptor binding inhibitors, and combinations thereof.
Compositions and methods of administration
The invention provides a pharmaceutical composition for activating latently infected AIDS virus, which contains the compound shown in the formula I or pharmaceutically acceptable salt thereof.
The pharmaceutical composition of the invention can be used for activating latent infected AIDS virus. When the pharmaceutical preparation of the present invention is used, other aids therapeutic agents, such as antiretroviral agents, may also be used simultaneously.
The pharmaceutical composition of the invention contains a safe and effective amount of the compound of the invention and a pharmaceutically acceptable carrier or excipient. Such vectors include (but are not limited to): saline, buffer, dextrose, water, glycerol, ethanol, powders, and combinations thereof. The pharmaceutical preparation should be compatible with the mode of administration.
The pharmaceutical composition of the present invention can be prepared in the form of an injection, for example, by a conventional method using physiological saline or an aqueous solution containing glucose and other adjuvants. Pharmaceutical compositions, such as tablets and capsules, can be prepared by conventional methods. Pharmaceutical compositions such as injections, solutions, tablets and capsules are preferably manufactured under sterile conditions. The pharmaceutical combination of the present invention may also be formulated as a powder for inhalation by nebulization. The amount of active ingredient administered is a therapeutically effective amount, for example, from about 1 microgram/kg body weight to about 50 mg/kg body weight, from about 5 microgram/kg body weight to about 10 mg/kg body weight, from about 10 microgram/kg body weight to about 5 mg/kg body weight per day. In addition, the compounds of the present invention may also be used with other therapeutic agents.
For the pharmaceutical compositions of the present invention, administration to a subject in need thereof (e.g., human and non-human mammals) can be by conventional means. Representative modes of administration include (but are not limited to): oral administration, injection, aerosol inhalation, etc.
In the case of pharmaceutical compositions, a safe and effective amount of the drug is administered to the mammal, wherein the safe and effective amount is generally at least about 10 micrograms/kg body weight, and in most cases does not exceed about 50 mg/kg body weight, preferably the dose is from about 10 micrograms/kg body weight to about 20 mg/kg body weight. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.
The main advantages of the invention include:
(a) provides a compound which can be used for activating latent infected AIDS virus;
(b) the compounds of the invention are low in cytotoxicity;
(c) the compounds of the invention have no non-specific immune activation.
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are by weight.
Experimental materials and research methods
1. Compounds to be detected
FICZ,25mM in DMSO; store at-80 ℃. FICZ (6-Formylindalo [3,2-b ]]carbazole) is a high affinity aromatic receptor AHR ligand derived from tryptophan. The structural formula is shown in figure 1, and the chemical formula is as follows: c19H12N2O。
2. Reagents and antibodies used
RPMI-1640 medium (GIBCO); fetal Bovine Serum (FBS) (Millipore); human lymphocyte isolate (Ficoll) (GE); dimethyl sulfoxide (DMSO) (Sigma); Phytohemagglutinin-P (PHA-P) (sigma); 3- (4, 5-dimethylthiazol-2) -2, 5-diphenyltetrazolium bromide salt (3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2-H-tetrazolium bromide, MTT) (cloudy days); trizol (invitrogen); THUNDERBIRD SYBR qPCR Mix (Toyobo); ReverTra qPCR RT Master Mix gDNA remove (Toyobo); FITC-anti CD69 (eBioscience); PE-anti CD25 (eBioscience); anti-CD4 antibody-embedded magnetic beads (Miltenyi).
HIV-1 latently infected Jurkat CD4+T cell model and latent activation assay
Using Jurkat CD4+T cells integrate HIV-1 genome to establish a latent infected cell model, and a GFP gene is inserted as a reporter gene, and cell cloning can be stimulated to activate and express GFP. Cells cultured in RPMI-1640/10% -FBS, 1X104Cells/well were seeded in 96-well cell culture plates, the compounds to be detected were added, and 24 hours later, the percentage of GFP positive cells was quantified by flow cytometry. Compound lytic reagent DMSO was used as a negative control for stimulation and epigenetic inhibitor SAHA was used as a positive drug control.
Resting (suppressing) CD4 of HIV-1 infected person+T cell isolation and virus activation detection
Recruitment of antiretroviral drugs to HIV-1 infected individuals who have been treated for more than 5 years and whose HIV-1 viral load cannot be detected in plasma. HIV-1 infected persons are recruited to follow-up outpatient service in the national base of TCM clinical in the first subsidiary hospital of the institute of TCM in Henan, to obtain medical ethical monographs, and have informed consent of the subjects.
Peripheral blood (50 mL) was collected, EDTA anticoagulated, and peripheral blood lymphocytes (PBMC) were isolated using a lymphocyte separation medium. Purification of CD4 Using anti-CD4 antibody-embedded magnetic beads+T cells. Cells were cultured in RPMI-1640/10% -FBS, 2X106Cells/well were seeded in 96-well cell culture plates. Adding a compound to be detected, staining Trypan Blue (Trypan Blue) after 48 hours, and counting the survival of cells; and collecting cell Trizol lysis extract RNA, reverse transcribing into cDNA by using ReverTra qPCR RT Master Mix gDNA remover, and amplifying HIV-1Gag gene on ABI Real-Time PCR instrument by using THUNDERBIRD SYBR qPCR Mix.
The primer sequences are as follows: gag forward 5'-GTGTGGAAAATCTCTAGCAGTGG-3' (SEQ ID NO.:1), Gag reverse 5'-CGCTCTCGCACCCATCTC-3' (SEQ ID NO.: 2); beta-actin-forward, 5'-GGGAAATCGTGCGTGACAT-3' (SEQ ID NO.:3), beta-actin-reverse, 5'-GTCAGGCAGCTCGTAGCTCTT-3' (SEQ ID NO.: 4).
5. Compound cytotoxicity assays
2x104Cells/well were seeded in 96-well cell culture plates, the compound to be tested was added, while a control group without compound was set, and after 24 hours, cell survival was tested using the MTT method: discarding 100. mu.L of culture medium/well, adding 20. mu.L/well of 5mg/mL MTT, and continuously incubating for 4h in a cell culture box; adding 100 mu L/hole DMSO, and standing overnight at 37 ℃ until the formazan is completely dissolved; the microplate reader detected OD570 and OD 630.
6. Compound immune activation assay
Healthy human peripheral blood PBMC cells, 2x10, were obtained by Ficoll density gradient centrifugation6The cells/well were seeded on 24-well cell culture plates, the compound to be tested was added while a control group without compound and PHA-P (5. mu.g/mL) was set, the cells were collected after 48 hours, stained with specific antibody, examined by flow cytometry, and CD4 was selected using anti-CD4-PE antibody (L3T 4; eBioscience)+T cells, cell surface CD25 and CD69 molecule expression was analyzed.
Example 1
FICZ activates HIV-1 latently infected Jurkat CD4+T cells
HIV-1 latently infected CD4 in human peripheral blood+The number of T cells is too low, every 100 ten thousand of CD4 +1 latently infected cell can be separated from T cell, which increases the difficulty of HIV-1 latently infection research. Currently, molecular mechanism research of HIV-1 latent infection, development of therapeutic means, and the like mainly utilize a cell model of latent infection.
Jurkat CD4 latently infected with HIV-1+T cells, finding that Compound FICZ significantly activates HIV-1 latency, EC50Up to 47nM, and the control drug SAHA EC50Then 700nM, compound FICZ showed nearly 15-fold activation compared to SAHA (figure 2). The results show that the compound FICZ activates HIV-1 latently infected Jurkat CD4+T cells.
Example 2
FICZ vs Jurkat CD4+T cells show lower cytotoxicity
To test the cytotoxicity of FICZ, different concentrations of FICZ were compared with HIV-1 latently infected Jurkat CD4+T cells were co-cultured for 48 hours and cell survival was examined using the MTT method. FICZ was found to have low cytotoxicity, and at FICZ concentrations as high as 50. mu.M, cells maintained cell viability as high as 82%.
Example 3
FICZ activates HIV-1 CD4 in resting infection+Replication in T cells
To validate the activation effect of FICZ on HIV-1 latent infection, HIV-1 infected subjects who were treated with antiretroviral drugs for more than 5 years and were unable to detect viral load in plasma were recruited, and CD4 was isolated as a resting infection of peripheral blood+T cells. Treatment of statically infected CD4 with Compound FICZ+After 48 hours on T cells, RNA was collected and HIV-1 replication was detected by quantifying the level of HIV-1gag mRNA. FICZ (100nM) treatment enhanced HIV-1 replication, CD4 isolated from 4 infected subjects+In T cells, the fold enhancement was 2.6-60 fold, respectively (FIG. 4). The results show that FICZ activates HIV-1 CD4 in the resting infection+Replication in T cells.
Example 4
FICZ resting CD4 for HIV-1 infected persons+T cells show lower cytotoxicity
To further test the cytotoxicity of FICZ, the statically infected CD4 purified from HIV-1 infected subjects was treated with FICZ+T cells, cells harvested after 48 hours, trypan blue staining, cell survival counted, FICZ (100nM) treatment found, CD4+T cells still maintained over 87% cell viability (figure 5). The results show that FICZ is silent to HIV-1 infected CD4+T cells showed lower cytotoxicity.
Example 5
FICZ vs. Primary CD4+T cells did not show non-specific immune activation
Abnormal activation of the immune system caused by HIV-1 infection can cause impairment of immune system function, and is an important factor for body pathogenesis. Thus, the activation of immune cells by FICZ was examined. PBMC cells from healthy human peripheral blood, high concentration FICZ (17,50,150 and500 μ M), specific antibody staining, flow cytometry detection, using anti-CD4-PE antibody (L3T 4; eBioscience) selection of CD4+T cell, analyzing cell surface CD25 and CD69 molecule expression to detect FICZ to CD4+Nonspecific activation of T cells. FICZ was found to be on primary CD4+T cells showed no non-specific immune activation (fig. 6).
Conclusion
Compound FICZ can activate HIV-1 latently infected Jurkat CD4+T cells, 15-fold more active than the clinical trial compound SAHA; treatment of isolated resting CD4 from HIV-1 infected persons with antiretroviral drugs+T cells, verifying that FICZ activates HIV-1 replication; FICZ exhibits low cytotoxicity; in addition, FICZ showed no non-specific activation of immune cells. In conclusion, the compound FICZ can efficiently activate HIV-1 latent infection and shows low cytotoxicity and no nonspecific immune activation; the FICZ can be combined with antiviral treatment, is used for eliminating latent HIV-1, and has clinical application value.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.
Figure IDA0001020081490000011
Figure IDA0001020081490000021

Claims (17)

1. The application of the compound shown in the formula I or the pharmaceutically acceptable salt thereof is characterized in that the compound is used for preparing a pharmaceutical composition or a preparation for activating the latent infected AIDS virus,
Figure FDA0003357464740000011
the compound of the formula I is a compound FICZ (6-formyl indolo [3,2-b ] carbazole).
2. The use of claim 1, wherein activating a latently infected HIV comprises activating HIV replication.
3. The use of claim 1, wherein the HIV comprises an HIV-1 type virus.
4. The use of claim 1, wherein the latent infection comprises a latent infection in an immune cell.
5. The use of claim 4, wherein said immune cells comprise T cells.
6. The use of claim 5, wherein said T cells comprise CD4+T cells.
7. The use of claim 1, wherein the latent infection comprises a quiescent infection.
8. The use of claim 1, wherein said pharmaceutical composition further comprises an antiretroviral drug.
9. The use of claim 1, wherein the pharmaceutical composition is in the form of an oral dosage form or an injectable formulation.
10. The use of claim 1, wherein the pharmaceutical composition comprises 0.001 to 99 wt% of the compound of formula I or a pharmaceutically acceptable salt thereof, based on the total weight of the composition.
11. A pharmaceutical composition, wherein the composition comprises: active ingredient (a) a compound of formula I, or a pharmaceutically acceptable salt thereof;
Figure FDA0003357464740000012
an active ingredient (b) an antiretroviral drug;
and (c) a pharmaceutically acceptable carrier;
the compound of the formula I is a compound FICZ (6-formyl indolo [3,2-b ] carbazole).
12. The pharmaceutical composition of claim 11, wherein the antiretroviral drug is selected from the group consisting of: an HIV reverse transcriptase inhibitor, an HIV integrase inhibitor, an HIV proteolytic enzyme inhibitor, an HIV-cell membrane fusion inhibitor, an HIV-co-acceptor binding inhibitor, or a combination thereof.
13. A kit, comprising:
(i) a first container, and an active ingredient (a) a compound of formula I, or a pharmaceutically acceptable salt thereof, or a medicament containing the active ingredient (a) contained in the first container;
(ii) a second container, and an active ingredient (b) an antiretroviral drug, or a drug containing an active ingredient (b), contained in the second container; and
(iii) instructions for the combined administration of active ingredient (a) and active ingredient (b) for the treatment of AIDS;
the compound of the formula I is a compound FICZ (6-formyl indolo [3,2-b ] carbazole).
14. The kit of claim 13, wherein the drugs in the first and second containers are a single formulation comprising active ingredient (a) and a single formulation comprising active ingredient (b).
15. The kit of claim 13, wherein the antiretroviral drug is selected from the group consisting of: an HIV reverse transcriptase inhibitor, an HIV integrase inhibitor, an HIV proteolytic enzyme inhibitor, an HIV-cell membrane fusion inhibitor, an HIV-co-acceptor binding inhibitor, or a combination thereof.
16. A method for activating a latently infected hiv in vitro, comprising the steps of:
(a) contacting a cell latently infected with an aids virus with a compound of formula I of claim 1, or a pharmaceutically acceptable salt thereof, thereby activating the latently infected aids virus.
17. The method of claim 16, wherein in step (a), the compound of formula I, or a pharmaceutically acceptable salt thereof, is added to a cell culture system and contacted with cells latently infected with hiv.
CN201610428916.6A 2016-06-16 2016-06-16 Compound for activating latent infected AIDS virus and application thereof in AIDS treatment Active CN107510694B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610428916.6A CN107510694B (en) 2016-06-16 2016-06-16 Compound for activating latent infected AIDS virus and application thereof in AIDS treatment

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610428916.6A CN107510694B (en) 2016-06-16 2016-06-16 Compound for activating latent infected AIDS virus and application thereof in AIDS treatment

Publications (2)

Publication Number Publication Date
CN107510694A CN107510694A (en) 2017-12-26
CN107510694B true CN107510694B (en) 2022-03-29

Family

ID=60720744

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610428916.6A Active CN107510694B (en) 2016-06-16 2016-06-16 Compound for activating latent infected AIDS virus and application thereof in AIDS treatment

Country Status (1)

Country Link
CN (1) CN107510694B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110960669B (en) * 2019-12-06 2022-01-21 中山大学 HIV-1 latent infection activator thiostrepton

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1338468A (en) * 2000-08-21 2002-03-06 上海博德基因开发有限公司 Polypeptide-human arylhydrocarbon receptor protein 15.75 and polynucleotide for coding it
EP1075256B1 (en) * 1998-05-05 2003-07-30 Institut National De La Sante Et De La Recherche Medicale (Inserm) Arylhydrocarbon receptor ligand antagonists
CN101466363A (en) * 2006-05-03 2009-06-24 西姆莱斯有限责任两合公司 Ah receptor antagonists
WO2010148350A1 (en) * 2009-06-18 2010-12-23 Cornell University Photoproducts of tryptophan, their synthesis and uses thereof
CN104023713A (en) * 2011-09-07 2014-09-03 德国癌症研究中心 Means and methods for treating and/or preventing natural ahr ligand-dependent cancer
CN107488635A (en) * 2017-09-22 2017-12-19 中国人民解放军第三军医大学第三附属医院 Monoclonal cell and its production and use

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8729058B2 (en) * 2009-10-27 2014-05-20 Michael Zasloff Methods and compositions for treating and preventing viral infections

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1075256B1 (en) * 1998-05-05 2003-07-30 Institut National De La Sante Et De La Recherche Medicale (Inserm) Arylhydrocarbon receptor ligand antagonists
CN1338468A (en) * 2000-08-21 2002-03-06 上海博德基因开发有限公司 Polypeptide-human arylhydrocarbon receptor protein 15.75 and polynucleotide for coding it
CN101466363A (en) * 2006-05-03 2009-06-24 西姆莱斯有限责任两合公司 Ah receptor antagonists
WO2010148350A1 (en) * 2009-06-18 2010-12-23 Cornell University Photoproducts of tryptophan, their synthesis and uses thereof
CN104023713A (en) * 2011-09-07 2014-09-03 德国癌症研究中心 Means and methods for treating and/or preventing natural ahr ligand-dependent cancer
CN107488635A (en) * 2017-09-22 2017-12-19 中国人民解放军第三军医大学第三附属医院 Monoclonal cell and its production and use

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"3-methylcholanthrene activates human immunodeficiency virus type 1 replication via aryl hydrocarbon receptor";Ohata, H等;《MICROBIOLOGY AND IMMUNOLOGY》;20031231;第47卷(第5期);参见标题和第368页左栏第3段 *
"Evidence for New Light-Independent Pathways for Generation of the Endogenous Aryl Hydrocarbon Receptor Agonist FICZ";Smirnova, A等;《CHEMICAL RESEARCH IN TOXICOLOGY》;20151219;第29卷(第1期);参见标题和摘要 *
"Tryptophan Metabolism Activates Aryl Hydrocarbon Receptor-Mediated Pathway To Promote HIV-1 Infection and Reactivation";Yan-Heng Zhou等;《AMERICAN SOCIETY FOR MICROBIOLOGY》;20191217;第10卷(第6期);参见摘要 *

Also Published As

Publication number Publication date
CN107510694A (en) 2017-12-26

Similar Documents

Publication Publication Date Title
AU2021228008B2 (en) Use of nucleoside compound in treatment of coronavirus infectious diseases
CA3048643A1 (en) Hiv immunotherapy with no pre-immunization step
CA3028982A1 (en) Hiv pre-immunization and immunotherapy
JP2018524335A (en) HIV preimmunization and immunotherapy
JP2013541010A5 (en)
JP2015508782A (en) Ingenol derivatives in reactivation of latent HIV virus
US20240115604A1 (en) Methods of manufacturing genetically-modified lymphocytes
Nair et al. Effect of methamphetamine on expression of HIV coreceptors and CC-chemokines by dendritic cells
US10765664B2 (en) Treatment of infectious diseases
CN107510694B (en) Compound for activating latent infected AIDS virus and application thereof in AIDS treatment
JP5283106B2 (en) Hepatitis C virus inhibitor
CN114246847B (en) Application of chalcone compounds in treatment of coronavirus infection
WO2019067977A1 (en) Use of senolytic agents to eliminate persistent hiv reservoirs
EP4153752A1 (en) Interferon- inducing oligonucleotide duplexes and methods of use
JP2020525459A (en) Treatment to eliminate HIV reservoir and reduce viral load
WO1997037661A1 (en) Preventive and remedy for viral infections
CN103054867B (en) Application of fangchinoline for preparing medicine for treating or preventing HIV
CN115607550A (en) Application of a kind of TLR7/8 agonist in inhibiting HIV
Molyer Yildirir SMAC Mimetics Sensitize HIV-Infected Cells to MG1-Mediated Death
CN113712963B (en) Application of BET inhibitor BMS-986158 in preparation of anti-AIDS drug
US11247996B2 (en) Small molecule inhibitors for transcription factors
WO2025098496A1 (en) Pharmaceutical composition and methods for treating hiv-1 infection
Blanco Regulation of HIV latency dynamics in monocytes and monocyte-derived macrophages
van der Ploeg et al. TNFα-producing CD4+ T cells dominate the SARS-CoV-2-specific T cell response in COVID-19 outpatients and are associated with durable antibodies
Frau Development of Interfering Strategies Against the Innate Immune Inhibition by the VP35 Ebola virus Protein, a Key Viral Target

Legal Events

Date Code Title Description
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
CP03 Change of name, title or address
CP03 Change of name, title or address

Address after: Building 2, No. 225, Chongqing South Road, Huangpu District, Shanghai, 200025

Patentee after: Shanghai Institute of Immunology and Infection, Chinese Academy of Sciences

Address before: No. 411, Hefei Road, Huangpu District, Shanghai 200025

Patentee before: INSTITUT PASTEUR OF SHANGHAI, CHINESE ACADEMY OF SCIENCES