Application of gnetum alcohol in treatment of adriamycin cardiomyopathy and diseases caused by adriamycin cardiomyopathy
By using terpineol to prepare drugs in various dosage forms, the progression of doxorubicin cardiomyopathy can be inhibited, solving the problem of poor treatment efficacy of doxorubicin cardiomyopathy in existing technologies, and providing a safe and effective treatment approach and drug option.
Patent Information
- Application Number
- CN202511552419.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-10-28
- Publication Date
- 2025-12-19
AI Technical Summary
In the existing technology, the treatment methods for doxorubicin cardiomyopathy are limited and the effects are not ideal. Although dextromethorphan has a cardioprotective effect, it poses a carcinogenic risk. There is an urgent need to develop safe and effective alternative drugs.
Using terpineol or its pharmaceutically acceptable salts, drugs for the prevention or treatment of doxorubicin cardiomyopathy are prepared by inhibiting cardiac atrophy, decreased cardiac function, reduced left ventricular ejection fraction, and myocardial fibrosis. Dosage forms include emulsions, microemulsions, and gels. In vitro and in vivo experiments are conducted to verify their efficacy.
Buying methimazole significantly improves cardiomyocyte vitality, restores cardiac structure and function, inhibits myocardial fibrosis, reduces drug side effects, provides a safe and effective treatment for doxorubicin-induced cardiomyopathy, and broadens the range of drug options.
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Figure CN121154601A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedicine, specifically relating to the application of terpineol in the treatment of doxorubicin cardiomyopathy and its related symptoms. Background Technology
[0002] Doxorubicin-induced cardiomyopathy (DIC) is a common and serious complication of anthracycline antitumor drugs such as doxorubicin. Its clinical features include cardiac atrophy, decreased cardiac function, and increased myocardial fibrosis. DIC is often progressive and irreversible, and currently available drugs are limited, mainly relying on reducing chemotherapy doses or combining with cardioprotective agents, but the efficacy remains unsatisfactory. Therefore, there is an urgent need to develop safe and effective new drugs for the prevention and treatment of DIC. Dexpromethazine is the only FDA-approved cardioprotective agent for combating anthracycline-induced cardiotoxicity; however, dexpromethazine has carcinogenic potential and may increase the risk of secondary malignancies in patients. Therefore, there is an urgent need to develop alternative treatment strategies or drugs to minimize DIC.
[0003] Gnetol is a natural small-molecule component derived from plants of the genus *Gnetol*, and it exhibits good biocompatibility. Gnetol is an organic compound with the chemical formula C2. 14 H 12 O4, with a molecular weight of 244.243 and CAS number 86361-55-9, is a white to light yellow solid powder. Metformin is primarily used as a candidate drug for treating COVID-19 and osteoporosis. Its mechanism of action involves inhibiting the activity of enzymes such as COX-1 and HDAC, and it also possesses anti-inflammatory and antioxidant properties. Currently, there is no clear research to support the efficacy of metformin in treating heart disease. Summary of the Invention
[0004] Purpose of the invention: In view of the problems existing in the prior art, the present invention provides the application of methoxyphen ...
[0005] The present invention also provides a pharmaceutical composition for the prevention or treatment of doxorubicin cardiomyopathy and the symptoms thereof.
[0006] Technical solution: In order to achieve the above objectives, the present invention relates to the use of methoxyphenoxyol or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of cardiomyopathy.
[0007] The use of the methoxyphenoxyol or its pharmaceutically acceptable salt in the preparation of a medicament for the prevention or treatment of doxorubicin cardiomyopathy.
[0008] The use of the methoxyphenoxyol or its pharmaceutically acceptable salt in the preparation of a medicament for the prevention or treatment of symptoms caused by doxorubicin cardiomyopathy.
[0009] The use of the aforementioned acetamipridol or its pharmaceutically acceptable salt in the preparation of a drug for the prevention or treatment of doxorubicin cardiomyopathy by inhibiting cardiac atrophy, decreased cardiac function, reduced left ventricular ejection fraction and increased myocardial fibrosis.
[0010] The use of the methoxyphenoxyol or its pharmaceutically acceptable salt in the preparation of a medicament for the prevention or treatment of doxorubicin cardiomyopathy by inhibiting the decrease in heart weight and the reduction in cardiomyocyte area.
[0011] The use of the aforementioned terpineol or its pharmaceutically acceptable salt in the preparation of drugs for the prevention or treatment of doxorubicin cardiomyopathy by reducing myocardial structural damage and improving cardiac function.
[0012] The pharmaceutical composition for doxorubicin cardiomyopathy of the present invention is characterized in that it comprises terpineol or a pharmaceutically acceptable salt thereof as the sole therapeutic ingredient or in combination with other pharmaceutical ingredients for treating doxorubicin cardiomyopathy, and also comprises raw materials, excipients or carriers required for formulation.
[0013] The dosage form of the pharmaceutical composition preferably includes emulsions, microemulsions, gels, solutions, tinctures, films, ointments, creams, or patches.
[0014] The use of the pharmaceutical composition described in this invention in the preparation of reagents or drugs for treating doxorubicin cardiomyopathy.
[0015] The use of the pharmaceutical composition described in this invention in the preparation of a medicament for the prevention or treatment of symptoms caused by doxorubicin cardiomyopathy.
[0016] This invention provides the application of metronidazole in the preparation of drugs for the prevention or treatment of doxorubicin-induced cardiomyopathy and its related symptoms. Through in vitro cell experiments and studies using a mouse model of doxorubicin-induced cardiomyopathy, this invention demonstrates that metronidazole can significantly improve cardiomyocyte viability and enhance cardiac structure and function. Furthermore, this invention finds that metronidazole has significant efficacy against doxorubicin-induced cardiomyopathy in cell and animal models, improving cardiac function and reducing myocardial tissue structural damage.
[0017] This invention applies Gnetol to directly treat rat-derived H9C2 cardiomyocytes in vitro, and finds that Gnetol treatment in vitro can significantly inhibit rat H9C2 cardiomyocyte damage; this invention applies Gnetol intraperitoneally to prevent or treat doxorubicin-induced cardiomyopathy in mice, and finds that Gnetol significantly alleviates doxorubicin cardiotoxicity and improves cardiac function in mice; the Gnetol described in this invention can be used to prepare drugs against doxorubicin cardiomyopathy, providing a new approach and means for the treatment of doxorubicin cardiomyopathy.
[0018] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:
[0019] 1. The gentamicin used in this invention has high safety, is naturally derived, and has low toxicity and side effects. In an doxorubicin-induced cardiomyopathy model, it can significantly improve cardiac morphology and functional indicators. It can be administered via multiple routes and has broad prospects for clinical application.
[0020] 2. This invention utilizes gentianol to treat rat H9C2 cardiomyocytes in vitro, significantly increasing cardiomyocyte viability. Intraperitoneal injection of gentianol to treat DIC mice increases left ventricular and heart weight, increases cardiomyocyte area, improves cardiac function, and effectively inhibits myocardial fibrosis in doxorubicin-induced cardiomyopathy mice.
[0021] 3. This invention, through in vitro and in vivo studies, has discovered that berberine possesses significant potential for treating doxorubicin cardiomyopathy and can be developed as a novel anti-doxorubicin cardiomyopathy drug, providing a new approach and method for its treatment. Simultaneously, this invention offers entirely new options and ideas for current anti-doxorubicin cardiomyopathy drugs, broadening the selection scope and contributing to the development of this technological field.
[0022] 4. This invention uses a lower therapeutic dose of methoxyphen ... Attached Figure Description
[0023] Figure 1 The image shows the viability of mouse cardiomyocytes after direct in vitro treatment with Gnetol; among which... This indicates that p < 0.001.
[0024] Figure 2 The figure shows the heart and body weight of mice in each group after Gnetol treatment for DIC. This indicates that p < 0.05; This indicates that p < 0.01; This indicates that p < 0.001.
[0025] Figure 3 The image shows the results of echocardiography in mice; This indicates that p < 0.05; This indicates that p < 0.01; This indicates that p < 0.001.
[0026] Figure 4 This is a diagram showing the results of Masson staining of mouse myocardial tissue; the blue area in the diagram represents the degree of fibrosis. Detailed Implementation
[0027] The present invention will be further described below with reference to specific embodiments and accompanying drawings.
[0028] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were purchased from conventional biochemical reagent companies.
[0029] Eight-week-old wild-type mice (WT) were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd. Normal mouse feed was purchased from Jiangsu Xietong Biotechnology Co., Ltd.
[0030] Gnetol was purchased from TCI AMERICA (G0371), CAS No. 86361-55-9.
[0031] Doxorubicin was purchased from MCE (HY-15142R), CAS No. 25316-40-9.
[0032] Dextromethorphan was purchased from MCE (HY-B0581), CAS No. 24584-09-6.
[0033] Example 1
[0034] Gnetol direct treatment in vitro to detect the viability of rat H9C2 cardiomyocytes
[0035] This embodiment establishes an adriamycin-induced myocardial injury model using rat H9C2 cardiomyocytes. H9C2 cardiomyocytes were seeded in DMEM high-glucose medium containing 10% FBS and 1% penicillin-streptomycin solution and cultured in a 37°C, 5% CO2 incubator. When cell confluence reached 80%-90%, the cells were digested with a 0.25% trypsin-0.02% EDTA mixture and passaged in 96-well plates, with 10,000 cells seeded per well. Cell grouping and treatment: H9C2 cardiomyocytes were divided into a control group, an adriamycin model group, and a pretreatment group using doxorubicin. Control group: 200 μL of fresh medium without the drug was added, and the cells were cultured for 26 h. Adriamycin-induced cardiomyocyte injury model group: 200 μL of fresh medium without the drug was added for pre-culture for 2 h, then the original medium was removed, and 200 μL of fresh medium containing 1 μM adriamycin was added, and the cells were cultured for another 24 h. Pretreatment group with gypsum: 200 μL of fresh culture medium containing different concentrations of gypsum (0.5 μM, 1 μM, 2 μM, 5 μM, 10 μM, 15 μM, 20 μM, 25 μM, 30 μM) was added and pre-cultured for 2 h, followed by the addition of 1 μM doxorubicin and continued culturing for 24 h. Six replicates were set for each concentration.
[0036] Cell viability assay: Cell viability was assessed using the CCK-8 assay. 10 μL of CCK-8 solution was added to each well and incubated at 37℃ in a 5% CO2 incubator for 1 h. The absorbance (OD) of each well was measured at 450 nm using a microplate reader. Cell viability (%) = (OD value of experimental group - OD value of blank group) / (OD value of control group - OD value of blank group) × 100%. Experiments confirmed that treatment with 1 μM doxorubicin for 24 h significantly reduced cell viability to 60%, meeting the modeling requirements.
[0037] Compared with the doxorubicin model group, pretreatment with different concentrations of terbinafine for 2 h, at 2 μM and above, significantly improved the survival rate of H9C2 cardiomyocytes after doxorubicin injury (P < 0.05 or P < 0.01). Specifically, the cell survival rate increased by approximately 10% in the 2 μM terbinafine pretreatment group and by approximately 20% in the 30 μM terbinafine pretreatment group. With increasing terbinafine concentration, cell survival rate showed a certain upward trend. Figure 1In summary, treatment with 1 μM doxorubicin for 24 h significantly reduced cell viability, establishing a doxorubicin-induced myocardial injury model and causing cardiomyocyte damage. Pretreatment with different concentrations of metronidazole (1 μM - 30 μM) for 2 h effectively improved the survival rate of H9C2 cardiomyocytes after doxorubicin injury, with the improvement ranging from 10% to 20%. This indicates that metronidazole has a certain protective effect against doxorubicin-induced H9C2 cardiomyocyte injury, and its protective effect may exhibit a certain concentration-dependent effect, providing experimental evidence for metronidazole as a potential cardioprotective drug.
[0038] Example 2
[0039] Effects of intraperitoneal injection of Gnetol on mouse heart
[0040] Eight-week-old male C57BL / 6J mice were randomly divided into: a control group (intraperitoneal injection of PBS once daily for 4 weeks); a doxorubicin-DOX model group (DIC): mice were intraperitoneally injected with DOX at a dose of 5 mg / kg once weekly for 4 weeks, totaling 20 mg / kg; a doxorubicin + gynostemma pentaphyllum group (GNE): mice were intraperitoneally injected with DOX at a dose of 5 mg / kg once weekly for 4 weeks, totaling 20 mg / kg; gynostemma pentaphyllum solution (3.75 mg / kg) was injected intraperitoneally 2 hours before the first DOX treatment once daily for 4 weeks; mice were allowed free access to food; and a doxorubicin + dextromethorphan positive control group (DEX): mice were intraperitoneally injected with dextromethorphan solution (50 mg / kg) 2 hours before the first DOX treatment once weekly for 4 weeks, totaling 200 mg / kg; mice in all groups had a normal diet during the treatment period.
[0041] Mice were housed in an SPF-grade animal facility for one month.
[0042] After 28 days of rearing, blood was collected from the eyes of mice. The mice were then euthanized by cervical dislocation, and the hearts were removed as quickly as possible. The surface moisture was dried, the heart weight was measured, and the heart-to-tibia length ratio (HW / TL) was calculated. The test results are shown below. Figure 2 .
[0043] from Figure 2 As can be seen, compared with the control group (CON), the heart weight of mice in the doxorubicin model group (DIC) was significantly reduced, indicating cardiac atrophy. In contrast, the heart weight of mice in the gentamicin group (GNE) was significantly increased compared with the doxorubicin model group, and the effect was better than the DEX positive control group even at a significantly lower dosage. This indicates that gentamicin can improve doxorubicin-induced cardiac atrophy, restore heart weight, and is safer to use at a lower dosage.
[0044] Example 3
[0045] Effects of intraperitoneal injection of Gnetol on cardiac function in mice
[0046] The mice treated in Example 2 were observed using echocardiography to assess their cardiac function. The steps for echocardiography are as follows:
[0047] ① The mice were shaved in the abdomen and chest cavity, and then anesthetized with isoflurane gas at a flow rate of 1.5-2%. The heart rate of the mice was kept stable at 380-400 beats / min before testing.
[0048] ②Using the Vevo3100 high-resolution in vivo imaging system (Vevo3100LT, Canada) for ultrasound examination. The heart was quickly located, and images and related parameters of the mouse heart's long and short axes were recorded in the B-model and M-model, respectively.
[0049] ③ The left ventricular ejection fraction (EF%) and left ventricular shortening fraction (FS%) were calculated using the Vevo3100 system. The results are as follows: Figure 3 .
[0050] from Figure 3 It can be seen that, compared with the heart of WT mice, the left ventricular ejection fraction and left ventricular shortening fraction of DIC mice were abnormally reduced, indicating cardiac systolic dysfunction. However, intraperitoneal injection of Gnetol significantly increased the left ventricular ejection fraction and left ventricular shortening fraction of DIC mice, basically restoring them to the level of the control group. The treatment effect was better than that of the positive control drug.
[0051] Example 4
[0052] Observation of cardiomyocyte size and tissue fibrosis in myocardial tissue
[0053] After euthanizing the mice treated in Example 2 by cervical dislocation, the mice were rapidly perfused systemically with 4% paraformaldehyde solution to flush out residual blood from the myocardial tissue. The mouse hearts were then removed and fixed overnight in 4% paraformaldehyde. The fixed hearts were then dehydrated, cleared, and embedded in paraffin. Paraffin sections (5 μm) were prepared after embedding.
[0054] The specific steps for the cardiac troponin (cTNT) immunofluorescence staining experiment are as follows:
[0055] ① Treat the slides with 0.2% Triton-X100 for 5 min, then wash with PBS 3 times (5 min each time).
[0056] ② Block with 5% BSA solution for 2 h, and wash with PBS 3 times (5 min / time) after blocking.
[0057] ③ Incubate cTNT antibody (1:400, diluted with PBS) at 4ºC overnight, remove the antibody, and wash 3 times with PBS (5 min / time).
[0058] ④ Incubate with fluorescent anti-rabbit secondary antibody (1:500, diluted with PBS) for 1 h. After incubation, remove the secondary antibody and wash with PBS 3 times (5 min / time).
[0059] ⑤ Incubate with DAPI (5 μg / ml, diluted with PBS) for 20 min. After incubation, remove the DAPI staining solution and wash with PBS 3 times (5 min each time).
[0060] ⑥ Finally, the slides were mounted with an anti-fluorescence quencher, and fluorescence images were taken using a laser confocal microscope (Carl Zeiss LSM880, Germany). The results are as follows: Figure 4 As shown above.
[0061] Masson staining of the slides was performed using the Masson trichrome staining kit (Sevier Biosciences, G1006), as follows:
[0062] ① Soak the slices in solution A at room temperature overnight (about 15 h).
[0063] ② Mix equal volumes of solution B and solution C (prepare fresh for immediate use), immerse the slices in the mixture of A and B for 1 min, rinse briefly with running water, and then differentiate for 1 min with 1% hydrochloric acid alcohol (concentrated hydrochloric acid: anhydrous ethanol = 1:100) until the cell nuclei are grayish-black and the background is almost colorless or light gray.
[0064] ③ Rinse briefly with running water, drain excess water from the sections, and immerse the sections in solution D for 6 minutes. At this point, the tissue will turn bright red. Drain the sections slightly (do not let them dry), and immediately immerse them in solution E for about 1 minute. This step is for differentiation. Differentiation continues until the collagen fibers turn light red and the fibers turn red, about 1-2 minutes.
[0065] ④ After slightly draining the E solution, immerse the slide directly in the F solution for 2-30 seconds without washing with water.
[0066] ⑤ The slices were washed and differentiated in three consecutive tanks of 1% glacial acetic acid for about 8 seconds each. Then they were dehydrated in three consecutive tanks of anhydrous ethanol for about 5 seconds, 10 seconds, and 30 seconds respectively. Finally, they were dehydrated in two tanks of n-butanol for 30 seconds and 2 minutes respectively.
[0067] ⑥ Finally, the slides were cleared with xylene in two baths, 5 minutes each time, then mounted with neutral resin. After complete drying, the slides were photographed under an optical microscope. The results are as follows: Figure 4 As shown below.
[0068] from Figure 4The results show that the area of cardiomyocytes in mice with doxorubicin cardiomyopathy was significantly smaller than that in normal mice, with a marked increase in myocardial fibrosis. Gnetol effectively increased the area of cardiomyocytes and inhibited the progression of myocardial fibrosis in DIC mice. This indicates that Gnetol has great potential for treating the early progression of doxorubicin cardiomyopathy.
[0069] In summary, this invention has demonstrated in in vitro experiments that Gnetol effectively inhibits the decline in the viability of mouse cardiomyocytes; and in in vivo studies, it has shown that Gnetol effectively inhibits the progression of doxorubicin cardiomyopathy and the occurrence of myocardial fibrosis. Both in vitro and in vivo experiments demonstrate that Gnetol significantly reduces disease progression in mice with doxorubicin cardiomyopathy, making it an effective target for the treatment of this disease. It can be developed as a novel anti-doxorubicin cardiomyopathy drug or drug target, providing a new approach and method for the detection and treatment of doxorubicin cardiomyopathy.
Claims
1. Use of methoxyphenoxyol or its pharmaceutically acceptable salts in the preparation of medicines for the prevention or treatment of cardiomyopathy.
2. The application according to claim 1, characterized in that, The use of the gentamicin or a pharmaceutically acceptable salt thereof is preferred in the preparation of a medicament for the prevention or treatment of doxorubicin cardiomyopathy.
3. The application according to claim 1, characterized in that, The use of the aforementioned terpineol or its pharmaceutically acceptable salt in the preparation of a medicament for the prevention or treatment of symptoms caused by doxorubicin cardiomyopathy.
4. The application according to claim 1, characterized in that, The use of the aforementioned terpineol or its pharmaceutically acceptable salt in the preparation of medicaments for the prevention or treatment of doxorubicin cardiomyopathy by inhibiting cardiac atrophy, decreased cardiac function, reduced left ventricular ejection fraction, and increased myocardial fibrosis.
5. The application according to claim 1, characterized in that, The use of the aforementioned terpineol or its pharmaceutically acceptable salt in the preparation of medicaments for the prevention or treatment of doxorubicin cardiomyopathy by inhibiting the decrease in heart weight and the reduction in cardiomyocyte area.
6. The application according to claim 1, characterized in that, The use of the aforementioned terpineol or its pharmaceutically acceptable salt in the preparation of medicaments for the prevention or treatment of doxorubicin cardiomyopathy by reducing myocardial structural damage and improving cardiac function.
7. A pharmaceutical composition for doxorubicin cardiomyopathy, characterized in that, It includes the use of gynostemma pentaphyllum or its pharmaceutically acceptable salt as the sole therapeutic ingredient or in combination with other ingredients of doxorubicin cardiomyopathy drugs, as well as the raw materials, excipients or carriers required for the formulation.
8. The pharmaceutical composition for treating hypertrophic cardiomyopathy according to claim 7, characterized in that, The dosage form of the pharmaceutical composition preferably includes emulsions, microemulsions, gels, solutions, tinctures, films, ointments, creams, or patches.
9. Use of the pharmaceutical composition of claim 7 in the preparation of a reagent or medicament for treating doxorubicin cardiomyopathy.
10. Use of the pharmaceutical composition of claim 7 in the preparation of a medicament for the prevention or treatment of symptoms caused by doxorubicin cardiomyopathy.