Application of cryptotanshinone in preparation of medicine for relieving toxic and side effects of osimertinib
Through the combination of cryptanshinone and osimirinib, the skin toxicity problem caused by osimirinib is solved, the skin toxicity symptoms are significantly improved, and the clinical application scope of osimirinib is expanded. The use of cryptanshinone is low and safe.
Patent Information
- Application Number
- CN202510450404.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-11
- Publication Date
- 2025-07-11
AI Technical Summary
The prior art lacks effective means to alleviate the skin toxic side effects caused by osimertinib, resulting in a decline in the quality of life of patients and interruption of treatment.
The combination of cryptanshinone and osimertinib was used to significantly inhibit the skin toxic reaction induced by osimertinib, including apoptosis of keratinocytes, DNA damage and inflammatory factors expression through in vitro cell experiments and in vivo experiments in mice.
It significantly reduces the skin toxicity caused by osimertinib, improves rash and desquamation, improves the safety and clinical application of osimertinib, is low in dose and high in safety, and is suitable for preparation of various dosage forms.
Smart Images
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of medicine, and specifically relates to the application of cryptotanshinone in the preparation of a drug for reducing the toxic and side effects of osimertinib, and is the application of cryptotanshinone in the preparation of a drug for treating the skin toxicity induced by the epidermal growth factor receptor inhibitor osimertinib. Background Art
[0002] Osimertinib (trade name Tagrisso) is an oral third-generation EGFR-TKI, which has significant efficacy against non-small cell lung cancer with EGFR mutations, especially has a special effect on the T790M drug-resistant mutation. It can irreversibly inhibit EGFR with EGFRm (Exon 19 deletion [Ex19del] / L858R) and EGFR T790M drug-resistant mutations, thus showing good efficacy clinically. Osimertinib shows a longer progression-free survival (PFS) than the first-generation EGFR-TKIs (such as erlotinib or gefitinib) in patients with advanced NSCLC positive for the EGFR T790M mutation.
[0003] In the FLAURA study, compared with standard EGFR-TKIs (erlotinib or gefitinib), osimertinib as first-line treatment significantly prolonged the median PFS to 18.9 months, while that of the control group was 10.2 months. Based on this research result, osimertinib was approved for first-line treatment, and more and more clinical guidelines recommend it as the first-line preferred option for NSCLC with sensitive EGFR mutations.
[0004] Compared with the first-generation and second-generation EGFR-TKIs, the severity of the skin toxicity of osimertinib is lower, but the incidence rate is still above 30%. Common skin adverse reactions include acneiform rash, pruritus, skin dryness and paronychia.
[0005] At present, there is a lack of effective prevention and treatment means for the skin toxicity induced by osimertinib. Clinically, only dose reduction or drug withdrawal can be adopted, which not only seriously affects the quality of life of patients, but also can lead to dose adjustment or treatment interruption, and ultimately results in tumor progression. Therefore, in-depth study of the molecular mechanism of skin toxicity caused by osimertinib and finding effective therapeutic drugs based on this have important practical significance for the clinical safe, effective and wide application of osimertinib.
[0006] Cryptotanshinone is one of the main active ingredients of Salvia miltiorrhiza. Research shows that cryptotanshinone has various effects such as anti-inflammatory, anti-tumor and organ protection, and has good therapeutic effects and high safety. However, there is currently no research indicating that it can be used to treat the toxic and side effects caused by osimertinib. Summary of the Invention
[0007] The object of the present invention is to provide the use of cryptotanshinone in the preparation of a drug for reducing the toxic and side effects of osimertinib, which is a drug capable of alleviating the toxic reactions induced by osimertinib and can reduce the skin toxic and side effects caused by the single use of osimertinib.
[0008] The CAS number of cryptotanshinone provided by the present invention is 35825-57-1, and its molecular formula is C 19 H 20 O3, and its molecular weight is 296.36.
[0009] The present invention is proved by in vitro cell experiments and in vivo mouse experiments that cryptotanshinone has a significant inhibitory effect on osimertinib-induced skin toxicity, can be used to treat osimertinib-induced skin toxicity, and thus expands the clinical application of osimertinib.
[0010] The skin toxic and side effects of osimertinib include: hair loss and skin rash in mice.
[0011] Furthermore, the skin toxic and side effects of osimertinib include: up-regulation of the expression level of c-PARP, an apoptosis-related protein of keratinocytes, and up-regulation of the expression of γ-H2A.X, a DNA damage marker.
[0012] Furthermore, the skin toxic and side effects of osimertinib include: up-regulation of the levels of TNFα and IL6.
[0013] Preferably, the dosage of cryptotanshinone used in the in vivo model is 10 mg / kg, 20 mg / kg, and 40 mg / kg. Research shows that these dosage levels are safe and effective dosage levels.
[0014] The drug provided by the present invention is prepared from cryptotanshinone or its pharmaceutically acceptable salt and a carrier.
[0015] The present invention provides an effective therapeutic drug for skin toxicity induced by EGFR inhibitors such as osimertinib. Research shows that compared with the group using osimertinib alone, the combined use of cryptotanshinone can significantly inhibit the apoptosis of keratinocytes in the epidermal tissue of mice and reduce the damage of the epidermal tissue of mice. In addition, the dosage of cryptotanshinone is low and has no effect on the survival of animals, the normal function of the skin, etc., indicating that cryptotanshinone as a protective agent is reasonable and safe and has high feasibility.
[0016] Preferably, the combined use of 50 mg / kg osimertinib and 10 mg / kg, 20 mg / kg, and 40 mg / kg cryptotanshinone is used.
[0017] The pharmaceutically acceptable carrier is a filler, a wetting agent, a binder, a disintegrant, or a lubricant.
[0018] The pharmaceutical dosage form is a liquid preparation or a solid preparation, including oral solid preparations, oral liquid preparations, injections, lyophilized powder injections, large volume infusions, patches, ointments, gels, soft capsules or suppositories.
[0019] The beneficial effects of the present invention are as follows:
[0020] (1) The present invention provides a new use of cryptotanshinone in the preparation of pharmaceutical preparations for treating osimertinib drug toxicity. Cryptotanshinone can significantly improve the rash, desquamation and inflammatory reactions caused by osimertinib. By combining cryptotanshinone with osimertinib, the toxic and side effects are effectively reduced, the safety of osimertinib is improved, and its clinical application is expanded.
[0021] (2) Cryptotanshinone has a low dosage, high safety and moderate price, and can be made into a suitable dosage form according to needs, with high clinical feasibility. Description of the Drawings
[0022] Figure 1 It shows the survival rate of keratinocytes reversed by cryptotanshinone reduced by osimertinib.
[0023] Figure 2 It shows that cryptotanshinone inhibits the apoptosis of keratinocytes induced by osimertinib, and the expression of apoptosis-related protein Cleaved PARP in keratinocytes is down-regulated after the combination of cryptotanshinone and osimertinib.
[0024] Figure 3 It is a representative diagram of cryptotanshinone down-regulating the DNA damage marker γ-H2A.X.
[0025] Figure 4 It shows that cryptotanshinone restores the levels of inflammatory factors TNFα and IL6 in keratinocytes.
[0026] Figure 5 It is a diagram of the appearance of the abdominal skin of a mouse. Cryptotanshinone improves the skin toxicity induced by osimertinib.
[0027] Figure 6 It shows that cryptotanshinone inhibits the histopathological tissue damage of the skin of mice caused by osimertinib, and the hematoxylin-eosin (H&E) staining of the skin tissue of mice. Detailed Embodiments
[0028] The present invention will be further described below in conjunction with specific embodiments. The following embodiments are only used to illustrate the present invention and are not used to limit the scope of application of the present invention. Without departing from the spirit and essence of the present invention, any modification or replacement of the methods, steps or conditions of the present invention belongs to the scope of the present invention.
[0029] Unless otherwise specified, the test methods used in the following examples are conventional methods; the materials, reagents, etc. used are reagents and materials that can be obtained from commercial sources unless otherwise specified.
[0030] The CAS number of cryptotanshinone is 35825-57-1, and its molecular formula is C 19 H 20 O3, with a molecular weight of 296.36, purchased from Macklin. Human keratinocyte HaCaT was purchased from the Cell Resource Center of the Institute of Basic Medicine, Chinese Academy of Medical Sciences & School of Basic Medicine, Peking Union Medical College. Osimertinib (CAS number: 1421373-65-0) was purchased from TargetMol.
[0031] Example 1: Using human keratinocyte HaCaT as the research object, 5 groups were set up, namely the control group, the osimertinib group, the low-dose cryptotanshinone treatment group, the medium-dose cryptotanshinone treatment group, and the high-dose cryptotanshinone treatment group. The dose of osimertinib was 0.5 μM, the doses of cryptotanshinone were 1, 2, and 4 μM, and the control group was the solvent dimethyl sulfoxide (DMSO). After co-administering for 48 h, the CCK8 staining was used to detect the survival rate of HaCaT cells. The calculation formula was: the survival rate of cells in the drug-administered group = (the average absorbance value of cells in the drug-administered group) / (the average absorbance value of cells in the control group) × 100%.
[0032] The experimental results are specifically described as follows: As Figure 1 shown, the survival rate of cells in the osimertinib group was 43.0% ± 10.5% (p < 0.001), the survival rate of cells in the 1 μM cryptotanshinone treatment group was 50.3% ± 9.1% (p = 0.6681), the survival rate of cells in the 2 μM cryptotanshinone treatment group was 69.6% ± 8.3% (p = 0.0090), and the survival rate of cells in the 4 μM cryptotanshinone treatment group was 84.8% ± 8.8% (p < 0.001). It shows that the combination of cryptotanshinone can significantly increase the survival rate of keratinocytes under the action of osimertinib ( Figure 1 ).
[0033] Example 2: Using human keratinocyte HaCaT as the research object, 5 groups were set up, namely the control group, the osimertinib group, the low-dose cryptotanshinone treatment group, the medium-dose cryptotanshinone treatment group, and the high-dose cryptotanshinone treatment group. The dose of osimertinib was 0.5 μM, the doses of cryptotanshinone were 1, 2, and 4 μM, and the control group was the solvent dimethyl sulfoxide (DMSO). After co-administering for 48 h, the Western Blot technique was used to detect the expression of apoptosis-related proteins in cells.
[0034] The specific experimental results are described as follows: The Western Blot results showed that after osimertinib acted on keratinocytes alone, the expression of apoptosis-related protein c-PARP was significantly up-regulated. After co-treatment with cryptotanshinone, the protein expression level of c-PARP in cells was significantly decreased, indicating that co-treatment with cryptotanshinone could significantly reverse osimertinib-induced apoptosis of keratinocytes( Figure 2 ).
[0035] Example 3: Using human keratinocyte HaCaT as the research object, 5 groups were set up, namely the control group, the osimertinib group, the low-dose cryptotanshinone treatment group, the medium-dose cryptotanshinone treatment group, and the high-dose cryptotanshinone treatment group. The dose of osimertinib was 0.5 μM, and the doses of cryptotanshinone were 1, 2, and 4 μM. The control group was the solvent dimethyl sulfoxide (DMSO). After co-administration for 48 h, the expression of the cell DNA damage marker γ-H2A.X was detected by Western Blot technology.
[0036] The Western Blot results showed that after osimertinib acted on keratinocytes alone, the protein expression of the DNA damage marker γ-H2A.X was significantly up-regulated. After co-treatment with cryptotanshinone, the protein expression level of the cell DNA damage marker γ-H2A.X was significantly decreased( Figure 3 ).
[0037] Example 4: Using human keratinocyte HaCaT as the research object, 5 groups were set up, namely the control group, the osimertinib group, the low-dose cryptotanshinone treatment group, the medium-dose cryptotanshinone treatment group, and the high-dose cryptotanshinone treatment group. The dose of osimertinib was 0.5 μM, and the doses of cryptotanshinone were 1, 2, and 4 μM. The control group was the solvent dimethyl sulfoxide (DMSO). After co-administration for 48 h, the expression levels of the cell inflammatory factors TNFα and IL6 were detected by ELISA technology.
[0038] After osimertinib acted on keratinocytes alone, the levels of TNFα and IL6 increased significantly. After co-treatment with cryptotanshinone, the expression levels of TNFα and IL6 decreased significantly( Figure 4 ).
[0039] Example 5: Fifty male C57BL / 6 mice, 6 - 8 weeks old and weighing 20 - 23 g, were randomly divided into 5 groups, namely the control group, the osimertinib group, the low - dose cryptotanshinone treatment group, the medium - dose cryptotanshinone treatment group, and the high - dose cryptotanshinone treatment group, with 10 mice in each group. They were continuously administered drugs by gavage for 6 weeks. The dose of osimertinib was 50 mg / kg / day, and the doses of cryptotanshinone were 10 mg / kg / day, 20 mg / kg / day, and 40 mg / kg / day. The control group was given the solvent cyclodextrin solution. On the 42nd day of drug administration, the mice were anesthetized, weighed, and their abdominal hair was removed. Then, the symptoms of skin toxicity reactions were observed and photographed. Subsequently, the mice were sacrificed and their abdominal skin tissues were taken,
[0040] In the group treated with osimertinib alone, obvious hair loss and rash appeared in the abdominal skin tissues of the mice. In the low - dose cryptotanshinone treatment group, there was slight hair loss in the abdominal skin of the mice. The above phenomena did not occur in the control group, the medium - and high - dose cryptotanshinone treatment groups, indicating that cryptotanshinone can significantly improve the skin hair loss and rash symptoms induced by osimertinib( Figure 5 ).
[0041] The results of H&E staining of the abdominal skin tissues of the mice showed that after administration of osimertinib, the epidermis of the mice was severely damaged, showing phenotypes such as karyopyknosis and hair follicle atrophy. Compared with the mice in the osimertinib group, after treatment with cryptotanshinone, the epidermal thickness of the mice returned to normal, the proportion of karyopyknosis decreased, and the hair follicles returned to normal( Figure 6 ).
Claims
1. Use of cryptotanshinone in the preparation of a drug for reducing the toxic and side effects of osimertinib, wherein the CAS number of cryptotanshinone is 35825-57-1.
2. The application according to claim 1, wherein Cryptotanshinone can reduce the rash, hair loss and inflammatory reaction caused by osimertinib, restore the epidermal thickness of mice, reduce the proportion of karyopyknosis, and restore the number of hair follicles.
3. The application according to claim 1, wherein Cryptotanshinone can reduce the apoptosis of keratinocytes and the release of inflammatory factors caused by osimertinib, down-regulate DNA damage markers, and increase the survival rate of keratinocytes.
4. The application according to claim 1, wherein The drug is prepared from cryptotanshinone or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier as a filler, wetting agent, binder, disintegrant or lubricant.
5. The application according to claim 1, characterized in that The pharmaceutical dosage form is a liquid preparation or a solid preparation.