CN111374975A - Medicinal application of harmine - Google Patents
Medicinal application of harmine Download PDFInfo
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- CN111374975A CN111374975A CN201811609823.9A CN201811609823A CN111374975A CN 111374975 A CN111374975 A CN 111374975A CN 201811609823 A CN201811609823 A CN 201811609823A CN 111374975 A CN111374975 A CN 111374975A
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- harmine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
- A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
Abstract
The invention discloses a medicinal application of harmine. The application is the application of harmine or pharmaceutically acceptable salts and esters thereof in preparing the medicine for promoting the proliferation of myocardial cells. Experiments prove that the harmine can promote the proliferation of rat cardiac muscle cells and can also promote the proliferation of human induced pluripotent stem cell induced cardiac muscle cells (hiPSC-derived cardiac muscle cells). Therefore, the harmine can be used for preparing medicaments for promoting the proliferation of myocardial cells, medicaments for treating or preventing heart diseases and other related fields, and provides a new medicament and a new treatment idea for treating or preventing heart diseases such as myocardial infarction.
Description
Technical Field
The invention belongs to the field of medicines, and particularly relates to a medicinal application of harmine.
Background
Cardiovascular diseases have become the first killer threatening human health currently, and 4000 million heart failure patients worldwide have become the main cause of human death. It has been found that in mammals, cardiomyocytes gradually lose proliferative capacity after adult life, and once myocardial infarction occurs, the loss of cardiomyocytes cannot be reversed. About 20-40 million myocardial cells exist in adults, about 25% of myocardial cells are lost within hours after the occurrence of myocardial infarction, the proliferation capacity of the remaining myocardial cells is very limited and is not enough to recover the systolic function of the heart, and finally, the heart failure of patients dies. To solve this problem, besides surgery, basic transformation studies mainly include finding cardiac stem cells or precursor cells, transplanting the cells to the myocardial infarction area by induced differentiation, but lack of sufficient molecular markers and low transplantation efficiency limit the use of this technology; the fibroblasts are transdifferentiated into cardiomyocytes using reprogramming techniques, or supplemented to the missing myocardium by means of promoting endogenous cardiomyocyte proliferation. The existing research shows that the heart of lower vertebrates such as zebra fish, salamander and the like has strong regeneration capacity, and newborn suckling mice also have certain regeneration capacity, and the regeneration capacity is mainly realized through myocardial cell proliferation induced by injury, and the capacity disappears after the adult. Adult hearts have not been considered to regenerate in the past, but extensive evidence suggests that there is a slow renewal of mammalian cardiomyocytes, and studies have found that neonatal cardiomyocytes are derived from existing myocardium. Therefore, an increasing number of scientists are interested in a strategy that induces the proliferation of endogenous cardiomyocytes. Therefore, the search for drugs capable of inducing the proliferation of endogenous cardiac muscle cells is of great significance for the treatment of cardiovascular diseases of human beings.
Currently, the main disclosed uses of Harmine (Harmine, CAS No.442-51-3) are to promote bone growth (US2013315965A1), promote browning of white fat cells, resist fat, glucose intolerance and insulin resistance caused by high fat diet (CN104224780A), treat obesity and obesity syndromes, such as type 2 diabetes (WO2010080756A2), treat kidney disease, cancer (WO2010036567A2) and peronophythora litchi (CN 105037350A). However, no report is made about the induction of endogenous cardiomyocyte proliferation by harmine.
Disclosure of Invention
The invention aims to provide a new application of harmine or pharmaceutically acceptable salts and esters thereof.
The structural formula of the Harmine (Harmine) CAS No.442-51-3 is shown as the formula I:
the new application of the harmine or the pharmaceutically acceptable salt and ester thereof provided by the invention is the application of the harmine in preparing the medicine for promoting the proliferation of the myocardial cells.
The cardiomyocytes can be mammalian cardiomyocytes. The mammal includes a human.
The invention also protects the application of the harmine or the pharmaceutically acceptable salt and ester thereof in preparing the medicaments for preventing and/or treating cardiovascular diseases.
Further, in some embodiments of the invention, the cardiovascular disease is heart disease resulting from loss, injury or death of cardiomyocytes.
Further, in some embodiments of the present invention, the above-mentioned heart diseases include, but are not limited to, myocardial infarction and cardiomyopathy resulting from loss, damage or death of other myocardial cells, and the like.
The medicine for promoting myocardial cell proliferation or preventing and/or treating cardiovascular diseases, which is prepared from harmine or pharmaceutically acceptable salt or ester thereof as an active ingredient, also belongs to the protection scope of the invention.
When necessary, one or more pharmaceutically acceptable carriers can be added into the medicine; the carrier includes diluent, excipient, filler, binder, wetting agent, disintegrating agent, absorption enhancer, surfactant, adsorption carrier, lubricant, etc. which are conventional in the pharmaceutical field.
The above medicine can be made into various forms such as injection, tablet, powder, granule, capsule, oral liquid, paste, cream, etc.; the medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.
The above drugs can be introduced into body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue by injection, spray, nasal drop, eye drop, penetration, absorption, physical or chemical mediated method; or mixed or coated with other materials and introduced into body.
Experiments prove that the harmine can promote the proliferation of rat cardiac muscle cells and can also promote the proliferation of human induced pluripotent stem cell (hiPSC-derived cardiac muscle cells). Therefore, the harmine can be used for preparing medicaments for promoting the proliferation of myocardial cells, medicaments for treating or preventing heart diseases and other related fields, and provides a new medicament and a new treatment idea for treating or preventing heart diseases such as myocardial infarction.
Drawings
FIG. 1 is a graph showing the effect of small molecule harmine on the proliferation of myocardial cells in neonatal rats.
FIG. 2 is a graph showing the effect of small molecule harmine on the proliferation of human induced pluripotent stem cell-induced cardiomyocytes (hiPSC-derived cardiomyocytes).
Detailed Description
The present invention is described below with reference to specific embodiments, but the present invention is not limited thereto, and any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The Harmine used in the examples below was a solution of Harmine in DMSO.
In the following examples "FBS" is fetal bovine serum.
The cTnT-mAG-hGeminin (1/110) virus used in the following examples was constructed as follows:
the cTnT myocardial specific promoter (shown as a sequence 1 in a sequence table) and mAG-hGeminin (1/110) (GenBank: NM-015895) are assembled and cloned on a pShuttle vector (Addgene 16402) by pEASY-Uni Seamless Cloning and Assembly Kit (all-type gold CU101-01), and then are subjected to enzyme digestion by a restriction enzyme PmeI to collect a linear plasmid; the linearized plasmid was cotransformed with pAdEasy1DNA (Addgene 16400)BJ5183 is made competent, recombinant plasmid is selected and amplified, and 293A cells are transfected with the recombinant plasmid (7.5 × 10 is the day before transfection)5293A cells were plated in a 60mm dish and cultured in DMEM medium containing 5% fetal bovine serum until the cell count reached 1.0-1.5 × 106Then, transfecting the cells with the recombinant plasmid by using a calcium phosphate coprecipitation method, removing a culture solution containing coprecipitation particles the next day after transfection, washing with a PBS buffer solution, subpackaging the cells into a 6-hole plate (3 ml of DMEM culture solution containing 5% fetal calf serum is added into each hole), and standing for 6 hours to allow the cells to adhere to the wall; covering the agarose after 6 hours for the formation of viral plaques (plaques should form within 10-21 days, adding agarose/DMEM mixture every 4-5 days or when the medium turns yellow); after obtaining the initial virus, the adenovirus is purified by 2-3 rounds of amplification and cesium chloride density gradient centrifugation.
Human induced pluripotent stem cell-induced cardiomyocytes (hiPSC-derived pluripotent stem cells) used in the following examples were prepared according to the methods described in the following references: cell Res.2017Aug; 27(8) 1002-1019.doi 10.1038/cr.2017.84.Epub 2017Jun 16.
The Goat anti-Rabbit IgG (H + L) Highly Cross-Adsorbed Secondary Antibody used in the examples described below was a Secondary Antibody manufactured by Invitrogen having the trade name of A32731, available under the trade name Goat anti-Rabbit IgG (H + L) high hly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488.
The Goat anti-Mouse IgG (H + L) highly cross-Adsorbed Secondary Antibody used in the examples described below was a Secondary Antibody manufactured by Lifetechnologies under the trade designation A21424 under the trade name Goat anti-Mouse IgG (H + L) highlyCryCross-Adsorbed Secondary Antibody, Alexa Fluor 555.
Example 1 in vitro assay of Harpagine-derived substance for promoting proliferation of myocardial cells in rats
(1) Culture of cardiac muscle cells of SD rat
Cardiomyocytes from 3-day-old SD rats were isolated and cultured in DMEM high-sugar medium (Hyclone) + 5% horse serum (GIBCO) in a 37-degree, 5% carbon dioxide incubator.
(2) Experimental grouping and processing
The myocardial cells of SD rats were isolated, and 5% horse serum (GIBCO) + DMEM high-sugar medium culture (Hyclone) was added, cytarabine (final concentration 20umol/L) was added to suppress the growth of non-myocardial cells, cells were infected with cTnT-mAG-hGeminin (1/110) virus (MOI value of virus infection: 100) after 48 hours of attachment, and then changed to DMEM containing 0.5% FBS after 24 hours, and the cells were subjected to the divided-dose treatment, as follows:
a. experimental groups: harmine small molecules were added (final concentration in medium 5. mu. mol/l) for 24 hours.
b. Blank control group: the same amount of DMSO as in the a experiment group was added.
c. Positive control group: FBS was added to a final concentration of 10%, and DMSO was added in an amount equivalent to that in experiment a.
(3) Test method
After the 3 groups are treated for 24 hours, nuclei are stained with hoechst fluorescent dye, and then photographed by a high content living cell analyzer (Molecular Device), and mAG-hGeminin (1/110) positive cardiomyocytes and total cell numbers are analyzed.
(4) Results
The test results are shown in FIG. 1. As can be seen from FIG. 1, mAG-hGeminin (1/110) positive cardiomyocytes increased 7-fold after banglargine treatment compared to the blank control group (0.5% FBS + DMSO) and 3.5-fold compared to the positive control group (10% FBS + DMSO).
Example 2 in vitro assay of Harpagine-derived cardiac myocytes (hiPSC-derived cardiac myocytes) proliferation
hiPSC-derived cardiac cytocytes were obtained, treated with Harmine (final concentration in medium 5 μmol/l) for 48H, RPMI1640+ DMSO as control (control), washed three times with PBS after 48H, fixed with 4% PFA (paraformaldehyde) for 15min at room temperature, washed three times with PBS, blocked with 1% BSA/PBS/0.1% Triton at room temperature for 1H, and 1H followed by primary antibodies Ctnt (1:200, abcam, ab8295), Phospho-Histone H3(Ser10) antibody (1:200, CST,9701S) overnight at 4 ℃, washed 4 times with 1% BSA/PBS/0.02% Tween, secondary antibodies: goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary antibody (1:300, Invitrogen, A32731), goat anti-mouse IgG (H + L) highly cross-adsorbed secondary antibody (1:300, Life technologies, A21424), left at room temperature for 1H, washed 3 times with PBS/0.02% Tween, stained for nuclei with DAPI, photographed with a high content viable cell analyzer (Molecular Device), and analyzed for the number of cardiomyocytes and total number of cells positive to PH 3.
The test results are shown in FIG. 2. As can be seen from fig. 2, the peganine-treated PH 3-positive human induced pluripotent stem cell-induced cardiomyocytes (hiPSC-derived cardiomyocytes) were increased nearly 3-fold compared to the control group.
Sequence listing
<110> Beijing university
Pharmaceutical application of harmine
<130>GNCZJ182500
<160>1
<170>PatentIn version 3.5
<210>1
<211>836
<212>DNA
<213> Chicken (Gallus domistiaus)
<400>1
tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg atcctctaga 60
actatagcta gaattcgccc ttacgggccc cccctcgagg tcgggataaa agcagtctgg 120
gctttcacat gacagcatct ggggctgcgg cagagggtcg ggtccgaagc gctgccttat 180
cagcgtcccc agccctggga ggtgacagct ggctggcttg tgtcagcccc tcgggcactc 240
acgtatctcc gtccgacggg tttaaaatag caaaactctg aggccacaca atagcttggg 300
cttatatggg ctcctgtggg ggaaggggga gcacggaggg ggccggggcc gctgctgcca 360
aaatagcagc tcacaagtgt tgcattcctc tctgggcgcc gggcacattc ctgctggctc 420
tgcccgcccc ggggtgggcg ccggggggac cttaaagcct ctgcccccca aggagccctt 480
cccagacagc cgccggcacc caccgctccg tgggacgatc cccgaagctc tagagcttta 540
ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct 600
cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa 660
gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt 720
tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc 780
actcccagtt caattacagc tcttaaggct agagtactta atacgactca ctatag 836
Claims (5)
1. Application of harmine or its pharmaceutically acceptable salt and ester in preparing medicine for promoting myocardial cell proliferation is provided.
2. Use according to claim 1, characterized in that: the cardiac muscle cell is a mammalian cardiac muscle cell.
3. Use according to claim 2, characterized in that: the mammal includes a human.
4. Application of harmine or pharmaceutically acceptable salt and ester thereof in preparing medicine for preventing and/or treating cardiovascular diseases is provided.
5. Use according to claim 4, characterized in that: the cardiovascular disease is heart disease caused by myocardial cell loss or death due to various reasons.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811609823.9A CN111374975A (en) | 2018-12-27 | 2018-12-27 | Medicinal application of harmine |
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| CN201811609823.9A CN111374975A (en) | 2018-12-27 | 2018-12-27 | Medicinal application of harmine |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112451529A (en) * | 2020-12-18 | 2021-03-09 | 忻佑康医药科技(南京)有限公司 | New pharmaceutical application of Kb-NB 142-70 |
| CN114133390A (en) * | 2021-12-20 | 2022-03-04 | 新疆医科大学 | Harmine derivative and preparation method and application thereof |
| CN114642662A (en) * | 2020-12-21 | 2022-06-21 | 忻佑康医药科技(南京)有限公司 | Novel application of MK-5046 medicine |
| CN114652717A (en) * | 2020-12-22 | 2022-06-24 | 忻佑康医药科技(南京)有限公司 | New application of naphazoline hydrochloride medicine |
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| DE19807993A1 (en) * | 1998-02-26 | 1999-09-02 | Bayer Ag | Treating tumor necrosis factor mediated inflammatory disease, e.g. arteriosclerosis, using new or known beta-carboline derivatives |
| CN100509810C (en) * | 2000-03-15 | 2009-07-08 | 萨诺费－阿文蒂斯德国有限公司 | Substituted beta-carbolines |
-
2018
- 2018-12-27 CN CN201811609823.9A patent/CN111374975A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19807993A1 (en) * | 1998-02-26 | 1999-09-02 | Bayer Ag | Treating tumor necrosis factor mediated inflammatory disease, e.g. arteriosclerosis, using new or known beta-carboline derivatives |
| CN100509810C (en) * | 2000-03-15 | 2009-07-08 | 萨诺费－阿文蒂斯德国有限公司 | Substituted beta-carbolines |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112451529A (en) * | 2020-12-18 | 2021-03-09 | 忻佑康医药科技(南京)有限公司 | New pharmaceutical application of Kb-NB 142-70 |
| CN114642662A (en) * | 2020-12-21 | 2022-06-21 | 忻佑康医药科技(南京)有限公司 | Novel application of MK-5046 medicine |
| CN114642662B (en) * | 2020-12-21 | 2023-08-25 | 忻佑康医药科技(南京)有限公司 | Pharmaceutical use of MK-5046 |
| CN114652717A (en) * | 2020-12-22 | 2022-06-24 | 忻佑康医药科技(南京)有限公司 | New application of naphazoline hydrochloride medicine |
| CN114652717B (en) * | 2020-12-22 | 2024-03-22 | 忻佑康医药科技(南京)有限公司 | Pharmaceutical application of naphazoline hydrochloride |
| CN114133390A (en) * | 2021-12-20 | 2022-03-04 | 新疆医科大学 | Harmine derivative and preparation method and application thereof |
| CN114133390B (en) * | 2021-12-20 | 2023-10-27 | 新疆医科大学 | Harmine derivative as well as preparation method and application thereof |
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Application publication date: 20200707 |