CN116327752A - Application of mulberry furan G in preparing melanoma treating medicine - Google Patents
Application of mulberry furan G in preparing melanoma treating medicine Download PDFInfo
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- CN116327752A CN116327752A CN202310454621.6A CN202310454621A CN116327752A CN 116327752 A CN116327752 A CN 116327752A CN 202310454621 A CN202310454621 A CN 202310454621A CN 116327752 A CN116327752 A CN 116327752A
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- YLQBMQCUIZJEEH-UHFFFAOYSA-N Furan Chemical compound C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 title claims abstract description 50
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- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 description 1
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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
- A61K31/335—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
- A61K31/352—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 condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Oncology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses an application of mulberry furan G in preparing a melanoma treatment drug. The invention discovers that the mulberry furan G has a certain effect of inhibiting the growth and metastasis of melanoma for the first time, and further researches the treatment effect and mechanism research of the mulberry furan G on the proliferation of the melanoma. The mulberry furan G can be used as an active ingredient and applied to medicines for treating metastatic melanoma, so that a new application of the mulberry furan G is developed, and a new choice is provided for treating metastatic melanoma.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of mulberry furan G in preparation of a melanoma treatment drug.
Background
Malignant melanoma is a malignant tumor which originates from melanocytes and has high invasiveness and drug resistance, and epidemiological data worldwide show that the incidence of malignant melanoma is third in skin malignant tumors, but the death rate of malignant melanoma is the first in skin malignant tumors. Melanoma is extremely poor in prognosis and easy to metastasize in early stage, and biological treatment, skin-directed treatment and radiotherapy are also adjuvant therapies applied to the treatment of melanoma in addition to chemotherapy. The traditional primary method of treating metastatic melanoma is chemotherapy with dacarbazine, temozolomide, fotemustine or taxanes, however, these chemotherapy methods do not increase patient survival. Since melanoma cells exhibit inherent resistance to chemotherapeutic drugs through a variety of mechanisms, other therapeutic strategies need to be sought. In recent years, immunotherapy has been developed in the field of melanoma treatment by improving the ability of the immune system to recognize and kill cancer cells. Currently, a large number of PD-1 antibodies, CTLA-4 monoclonal antibodies, interferon (IFN) and the like are used.
The mulberry furan G is a flavonoid compound extracted from mulberry, and the structural formula is shown in figure 1. Modern pharmacological researches show that mulberry branches and barks contain a large amount of bioactive components, have important pharmacological characteristics, and subsequently find active components with functions of reducing blood sugar, blood fat and the like, so that the research on the phytochemical components is more and more important. However, the research on whether the mulberry furan G has anti-tumor effect is very little at present.
Disclosure of Invention
The invention aims to provide an application of mulberry furan G in preparing a melanoma treatment drug.
The structural formula of the mulberry furan G is shown as the following formula:
in order to achieve the above purpose, the present invention adopts the following technical means:
application of Mortiered G in preparation of medicine for preventing and treating melanoma is provided.
Further, the mulberroside G can treat metastatic melanoma by inhibiting proliferation of melanoma cells.
Furthermore, the mulberry furan G can further treat metastatic melanoma by inhibiting the migration of melanoma cells.
Furthermore, the mulberry furan G can further treat metastatic melanoma by inhibiting the expression of melanoma cell immune checkpoints.
Furthermore, the mulberry furan G can further treat metastatic melanoma by inhibiting the transformation of melanoma cell epithelium and stroma.
The invention is to find the effect of the mulberry furan G in inhibiting the transfer of melanoma for the first time, and can be developed as a chemotherapy means for preventing and treating the melanoma by using the mulberry furan G alone or in combination.
The mulberry furan G provided by the invention can be used as an active ingredient, is applied to medicines for treating metastatic melanoma, exploits a new application of the mulberry furan G, and provides a new choice for treating metastatic melanoma.
Drawings
FIG. 1 shows the effect of Mortiered furang on inhibition of B16F10 cell proliferation.
FIG. 2 shows the effect of Mulberry G on A375 cell proliferation inhibition.
FIG. 3 shows the effect of Mortiered G on B16F10 cell scratch repair.
FIG. 4 shows the effect of Mulberry G on A375 cell scratch repair.
FIG. 5 is the effect of Mortiered G on PD-1 expression in A2058 cells.
FIG. 6 is the effect of Mulberry G on A2058 cell epithelial-mesenchymal transition.
Detailed Description
The invention will now be described in further detail with reference to the drawings and specific examples, which should not be construed as limiting the invention. Modifications and substitutions to methods, procedures, or conditions of the present invention without departing from the spirit and nature of the invention are intended to be within the scope of the present invention. The experimental procedures and reagents not shown in the formulation of the examples were all in accordance with the conventional conditions in the art.
Example 1
CCK8 experiment
When murine melanoma cells B16F10 and human melanoma cells a375 grew to log phase, the cells were digested with trypsin, washed to blow off, and seeded into 96-well plates at 5000 cells per well with a volume of 100 μl per well. Cells were inoculated 24 hours in advance for CCK8 experiments. Mulberry furan G was dissolved in DMSO solution to prepare a mother solution at a concentration of 10 Mmol/L. After the cells were attached, they were randomly divided into a control group and a group of different concentrations of Mortierella mulberrosins G (final concentrations of 0.9375, 2.5, 3.75, 5, 10, 15, 30, 40, 60. Mu. Mol/L), and a blank group containing no cells but DMEM complete medium was provided, with 3 multiplex wells per group. 100 mu L of DMEM complete medium is added into a control group, 100 mu L of complete medium containing corresponding concentration of medicines is added into each concentration group of the mulberry furan G respectively, 10 mu L of CCK8 solution is added into each group of cells when the cells are cultured for 24 hours, after incubation is carried out for 2 hours at 37 ℃, absorbance (OD) of each hole is measured at a wavelength of 450nm by adopting an enzyme-labeling instrument, and the cell survival rate is calculated:
as can be seen from fig. 1 and 2, the mulberry furan G showed significant cell proliferation inhibitory activity against both murine melanoma cells B16F10 and human melanoma cells a 375. The mulberry furan G has obvious inhibition effect on the activity of murine melanoma cells B16F10 and human melanoma cells A375 within the range of 0.9375-60 mu mol/L and is concentration dependent.
Example 2
Scratch repair experiment
Taking murine melanoma cells B16F10 and human melanoma cells A375 in a logarithmic growth phase, inoculating the murine melanoma cells B16F10 and the human melanoma cells A375 in a 6-well plate, and carrying out cell streaking on the surface layer of single cells by using a 200 mu L sterile gun head until the cells adhere to the wall and reach 70% -80% in the next day.
After the scratch, the cells were washed 3 times with sterile PBS, and replaced with fresh serum-free DMEM medium containing 5. Mu.M of Mortierella xylostella G, and placed in an incubator for culturing for 24 hours. Cells were removed at 0h and 24h during the 24h incubation period, and cell scarification was observed under a microscope and photographed. The image J software was used to calculate the distance area between cell scratches and the cell migration efficiency.
As shown in FIGS. 3 and 4 (Scale bar:50 μm), in the system containing 5. Mu.M of Mulberry Furan G, the migration ability of murine melanoma cells B16F10 and human melanoma cells A375 was decreased compared to the solvent set. The above results demonstrate that mulberrosine G inhibits the ability of murine melanoma cell B16F10 and human melanoma cell A375 to migrate.
Example 3
Real-time quantitative PCR experiments
Inoculating human melanoma cells A2058 in logarithmic growth phase into cell culture dish, changing fresh culture medium containing 5 μm Mortierella dichotoma G when the cells adhere to wall, and culturing in incubator for 24 hr. Sample cells were collected, washed 2 times with pre-chilled PBS, 1mL Trizol was added, and the mixture was allowed to stand for 10min. Taking cell lysate, transferring the cell lysate into an enzyme-free 1.5mL centrifuge tube, adding 0.2mL of chloroform, stirring and mixing uniformly by vortex, standing for 5min, and centrifuging at 12000rpm for 15min. The supernatant was taken into a new enzyme-free 1.5mL centrifuge tube, added with an equal volume of isopropanol, gently inverted, allowed to stand for 10min, and centrifuged at 12000rpm for 10min. The supernatant was discarded, 1mL of absolute ethanol was added, and the mixture was centrifuged at 12000rpm for 5min. The absolute ethanol was discarded, left at room temperature until the ethanol evaporated, and 30. Mu.L of absolute water was added for dissolution. RNA concentration and purity were measured. Subsequently, reverse transcription was performed according to the instructions of the reverse transcription kit of the biological company of Nanjinopran. After cDNA was obtained, the cDNA was subjected to a real-time fluorescent quantitative PCR assay kit (America Biolabs, nanjinouzan) to obtain a relative quantification using the DeltaCT method, and GAPDH was used as an internal reference gene.
mRNA expression levels of human melanoma cell A2058 immune checkpoints and genes related to the epithelial-mesenchymal transition process are detected by a real-time quantitative PCR experiment. As shown in FIG. 5, mRNA expression levels of immune checkpoint PD-1 were down-regulated 24h after 5. Mu.M of SANGFUG treatment of human melanoma cells A2058. As shown in FIG. 6, 24 hours after 5. Mu.M of SANGFUG treatment of human melanoma cells A2058, the mRNA expression level of the epithelial-mesenchymal transition marker VIMENTIN was decreased, and the mRNA expression level of growth factor VEGF regulating the epithelial-mesenchymal transition was decreased. Taken together, the mulberroside G may inhibit the expression and epithelial-mesenchymal transition processes of human melanoma cell a2058 immune checkpoint, further inhibiting its invasion and migration.
Claims (4)
1. Application of Mortiered G in preparation of medicine for treating and/or preventing melanoma is provided.
2. Use of samaraofuran G in the manufacture of a medicament for the treatment and/or prevention of metastasis of melanoma in vivo.
3. Use according to claim 1 or 2, characterized in that: the mulberry furan G is used as the only active ingredient or one of active ingredient groups in the medicine.
4. Use according to claim 1 or 2, characterized in that: the medicament also includes a pharmaceutically acceptable excipient.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202310454621.6A CN116327752A (en) | 2023-04-25 | 2023-04-25 | Application of mulberry furan G in preparing melanoma treating medicine |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202310454621.6A CN116327752A (en) | 2023-04-25 | 2023-04-25 | Application of mulberry furan G in preparing melanoma treating medicine |
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2023
- 2023-04-25 CN CN202310454621.6A patent/CN116327752A/en active Pending
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