Combined pharmaceutical composition for treating osimertinib drug-resistant non-small cell lung cancer
Through the combined use of JK184 and osimertinib, the problem of drug resistance of non-small cell lung cancer after osimertinib treatment was solved, efficient tumor suppression rate was achieved, and toxicity was avoided, and a new treatment plan was provided. Wide clinical application potential.
Patent Information
- Application Number
- CN202411485693.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-23
- Publication Date
- 2024-12-27
AI Technical Summary
Patients with non-small cell lung cancer often develop drug resistance after osimertinib. The existing combination treatment methods have problems with low safety and less efficacy as expected. New drug combinations are needed for treatment.
The combination of JK184 and osimertinib is used, with a molar ratio of 1:100, including pharmaceutically acceptable pharmaceutical excipients, prepared in the form of water infusion, powder, etc., for the treatment of osimertinib resistance. Cell lung cancer.
The combined drug composition has a suppression rate of drug-resistant tumors of more than 95%, and does not produce toxicity higher than that of osimertinib alone, providing an effective treatment plan and having broad clinical application prospects.
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Figure CN119185315A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of medical technology and specifically relates to a combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer. Background Art
[0002] Non-small cell lung cancer (NSCLC) accounts for 85% of all lung cancer cases. It can be divided into three major subtypes based on morphological classification: large cell lung cancer, squamous cell carcinoma, and adenocarcinoma. The discovery of activating mutations in the gene encoding epidermal growth factor receptor (EGFR) tyrosine kinase in patients with non-small cell lung cancer (NSCLC) and the subsequent development of targeted therapy using small molecule EGFR tyrosine kinase inhibitors (TKIs) have greatly changed the treatment landscape for these tumors.
[0003] Somatic activating mutations in the EGFR tyrosine kinase domain are present in approximately 15% of Caucasians and nearly 50% of Asian patients with advanced non-small cell lung cancer. Almost 90% of these mutations include deletions in exon 19 or L858R point mutations within exon 21. These genetic changes act as oncogenic drivers, leading to ligand-independent activation of EGFR downstream signaling, thereby promoting cell proliferation, survival, and migration.
[0004] Although the survival rate of patients has been significantly improved after TKI treatment, most patients have developed drug resistance. The first and second generation TKIs (such as gefitinib) produced T790M mutations after patients took the drugs, while the third generation TKI-osimertinib overcomes the drug resistance caused by T790M mutations of the first and second generation TKIs and is currently the first and second line drug for the clinical treatment of NSCLC patients with EGFR mutations.
[0005] The molecular formula of JK184 is C19H18N4OS, and the molecular weight is 350.44. This small molecule is an effective Hedgehog (Hh) inhibitor. The preliminary research results of the inventor team of the present invention confirmed that JK184 can promote ferroptosis of osimertinib-resistant cell lines and aggravate the degree of autophagy of osimertinib-resistant cell lines.
[0006] Although osimertinib has a strong clinical activity, patients will inevitably develop secondary resistance to this treatment; at the same time, clinically, combined treatments for patients with drug-resistant non-small cell lung cancer, such as chemotherapy combined with targeted therapy, immunotherapy combined with targeted therapy, and the combined use of multiple targeted drugs, have low safety, poor patient prognosis, and unsatisfactory efficacy. Therefore, it is urgent to develop new drug combinations to treat osimertinib-induced resistance. Summary of the invention
[0007] In view of the deficiencies in the prior art, the object of the present invention is to provide a combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer, and to provide a drug combination scheme for the treatment of osimertinib-resistant non-small cell lung cancer.
[0008] To achieve the above object, the present invention adopts the following technical solutions: A combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer, the pharmaceutical composition comprising JK184 and osimertinib.
[0009] Preferably, the molar ratio of JK184 to osimertinib in the pharmaceutical composition is 1:100.
[0010] Furthermore, the pharmaceutical composition also includes a pharmaceutically acceptable pharmaceutical excipient, which is a pharmaceutically acceptable salt, excipient or carrier.
[0011] Preferably, the carrier is any one or more of a filler, a wetting agent, a binder, a disintegrant or a lubricant.
[0012] Furthermore, the pharmaceutical composition is in the form of any one or more of an aqueous extract, a powder, a lotion, a tincture, an oil, an emulsion, an ointment, a plaster or an aerosol.
[0013] Another object of the present invention is to provide a use of JK184 in combination with osimertinib in the preparation of a medicament for treating osimertinib-resistant non-small cell lung cancer.
[0014] To achieve the above object, the present invention adopts the following technical solutions: The use of JK184 in combination with osimertinib in the preparation of a drug for the treatment of osimertinib-resistant non-small cell lung cancer.
[0015] The beneficial effects of the present invention are: The present invention provides a combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer, and provides a drug combination scheme for treating osimertinib-resistant non-small cell lung cancer. The combined pharmaceutical composition comprises JK184 and osimertinib, and the combination of JK184 and osimertinib has a synergistic effect, so the combined pharmaceutical composition has an inhibition rate of more than 95% on drug-resistant tumors, and does not produce toxicity higher than that of osimertinib alone. The experimental results of the present invention provide a theoretical basis for the clinical combined use of osimertinib combined with JK184 and an effective scheme for treating osimertinib-resistant non-small cell lung cancer, and have broad application prospects in the field of medicine.
[0016] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, some of the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0018] Figure 1 : The inhibitory effect of JK184 combined with osimertinib on H1975OR cells; Figure 2 : Synergistic index of JK184 and osimertinib combined against H1975OR cells; Figure 3 : Actual photos of tumors in HCC827OR-inoculated tumor-bearing mice after combined use of JK184 and osimertinib; Figure 4 : Tumor volume change curve of HCC827OR-inoculated tumor-bearing mice after combined use of JK184 and osimertinib; Figure 5 : Body weight change curve of HCC827OR-inoculated tumor-bearing mice after combined use of JK184 and osimertinib. DETAILED DESCRIPTION
[0019] In order to better describe the present invention, the following is further illustrated by specific examples. The methods in the following examples are conventional methods unless otherwise specified.
[0020] The technical solutions described in the present invention, unless otherwise specified, are all conventional solutions in the art; the reagents or materials described, unless otherwise specified, are all from commercial channels.
[0021] 1.1 Experimental Materials: JK184 (MCE, HY-13307), osimertinib (MCE, HY-15772), CCK8 kit (AbMole, M4839).
[0022] HCC827 osimertinib-resistant cells (HCC827OR) and H1975 osimertinib-resistant cells (H1975OR) were constructed by our laboratory through increasing concentration gradients. The specific operation method is: the IC50 of the parental HCC827 and H1975 is determined by the CCK8 method, and half of the IC50 concentration is selected as the starting concentration to treat the parental cells. After two stable passages, the concentration is doubled; after the drug-resistant concentration exceeds 100nM, it is increased by 100nM; after the drug-resistant concentration exceeds 1μM, it is increased by 1μM. Finally, the experiment is carried out with drug-resistant cells that can grow stably at a concentration of 2μM.
[0023] Fetal bovine serum, brand Gibco; cell culture medium, RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin.
[0024] 1.2 Experimental method: CCK8 method to determine cell proliferation inhibition rate HCC827, H1975 and their drug-resistant cells in the logarithmic growth phase were taken, digested, digested, resuspended after centrifugation, and inoculated into 96-well plates. Each well contained 3x103 cells in 100μl complete medium. The plates were cultured in a cell culture incubator at 37°C and 5% CO2, and the drugs were added after 24h.
[0025] Experimental group 1: different concentrations of JK184 were added to make the concentrations in 100 μl culture medium 0, 1, 2, 5, 10, 15, 20, 30, 50 nM / L respectively; Experimental group 2: different concentrations of osimertinib were added to make the concentrations in 100 μl of culture medium 0, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20 μM / L respectively; Experimental Group 3: different concentrations of combined drugs were added, 1nM / LJK184+0.1μM / L Osimertinib, 2nM / LJK184+0.2μM / L Osimertinib, 5nM / LJK184+0.5μM / L Osimertinib, 10nM / LJK184+0.5μM / L Osimertinib, 20nM / LJK184+1μM / L Osimertinib, 15nM / LJK184+1μM / L Osimertinib, 20nM / LJK184+2μM / L Osimertinib, 30nM / LJK184+5μM / L Osimertinib, 50nM / L JK184+20μM / L Osimertinib; 0.1% DMSO was added to the control group and the culture medium was used as a blank control.
[0026] After 72 hours of action, 10 μl CCK8 was added to each well and placed in an incubator for 1.5 hours. The 96-well plate was taken out and the absorbance (OD) was measured at 450 nm using an ELISA reader. The inhibition rate of cell proliferation was calculated according to the following formula: Proliferation rate = (OD value of the drug-treated group - OD value of the blank group) / (OD value of the control group - OD value of the blank group) × 100%, inhibition rate = 1-proliferation rate.
[0027] CompuSyn software was used to calculate the combination index CI. The names of different drug combinations, concentration gradients and inhibition rates were entered into the software and calculated using the Chou-Talalay formula built into the software. When CI ≤ 0.9, the two drugs were considered to have a potential synergistic effect. The above steps were repeated three times for statistical analysis, and statistical difference analysis was performed in GraphPad Prism 8 software. If p < 0.05, it indicated a significant difference.
[0028] 1.3 Experimental Results The results of the CCK8 experiment are shown in Tables 1 and Figure 1~2 As shown: JK184 or osimertinib alone can inhibit the proliferation of H1975OR cells to a certain extent. The combined use of JK184 and osimertinib significantly enhanced the inhibitory effect on H1975OR cells, achieving a synergistic effect.
[0029] Table 1 Inhibitory effect of combined use of JK184 and osimertinib on H1975OR cells
[0030] 2.1 Experimental Materials: JK184 (MCE, HY-13307), osimertinib (MCE, HY-15772), CCK8 kit (AbMole, M4839).
[0031] HCC827 osimertinib-resistant cells (HCC827OR) and H1975 osimertinib-resistant cells (H1975OR) were constructed by our laboratory through increasing concentration gradients. The specific operation method is: the IC50 of the parental HCC827 and H1975 is determined by the CCK8 method, and half of the IC50 concentration is selected as the starting concentration to treat the parental cells. After two stable passages, the concentration is doubled; after the drug-resistant concentration exceeds 100nM, it is increased by 100nM; after the drug-resistant concentration exceeds 1μM, it is increased by 1μM. Finally, the experiment is carried out with drug-resistant cells that can grow stably at a concentration of 2μM.
[0032] Fetal bovine serum, brand Gibco; cell culture medium, RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin.
[0033] Four-week-old male nude mice were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.
[0034] 2.2 Experimental methods: Testing the effect of JK184 combined with osimertinib in tumor-bearing mice Four-week-old male nude mice were housed in an SPF environment for one week, and then 4×10 6 HCC827ORs were inoculated into the back of each mouse using a disposable sterile syringe.
[0035] One week later, the mice were given drugs by intraperitoneal injection and divided into 1 control group and 3 experimental groups, with 5 mice in each group. Experimental group 1 was injected with J1K84 at 5 mg / kg. Experimental group 2 was injected with osimertinib at 5 mg / kg, and experimental group 3 was injected with JK184 and osimertinib at 5 mg / kg. The control group was injected with an equal amount of solvent. The weight and tumor volume of mice were recorded once a week. The length and width of the tumor volume were measured with a vernier caliper. Tumor volume = (length x width2) / 2.
[0036] After 8 weeks of administration, the mice were euthanized, the tumors were removed and photographed, and GraphPadPrism 8 was used to statistically analyze the change curves of mouse body weight and tumor volume.
[0037] 2.3 Experimental Results The results of the tumor-bearing mouse experiment were as follows Figures 3-5 As shown: JK184 or osimertinib alone can inhibit the proliferation of HCC827OR cells in nude mice to a certain extent. The combined use of JK184 and osimertinib significantly enhanced the inhibitory effect on HCC827OR cells, achieving a synergistic effect.
[0038] The present invention provides a combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer, and provides a drug combination scheme for treating osimertinib-resistant non-small cell lung cancer. The combined pharmaceutical composition comprises JK184 and osimertinib, and the combination of JK184 and osimertinib has a synergistic effect, so the combined pharmaceutical composition has an inhibition rate of more than 95% on drug-resistant tumors, and does not produce toxicity higher than that of osimertinib alone. The experimental results of the present invention provide a theoretical basis for the clinical combined use of osimertinib combined with JK184 and an effective scheme for treating osimertinib-resistant non-small cell lung cancer, and have broad application prospects in the field of medicine.
[0039] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.
[0040] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.
Claims
1. A combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer, characterized in that: The pharmaceutical composition comprises JK184 and osimertinib.
2. The combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer according to claim 1, characterized in that: The molar ratio of JK184 to osimertinib in the pharmaceutical composition is 1:
100.
3. The combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer according to claim 1, characterized in that: The pharmaceutical composition further comprises a pharmaceutically acceptable pharmaceutical excipient, which is a pharmaceutically acceptable salt, excipient or carrier.
4. The combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer according to claim 3, characterized in that: The carrier is any one or more of a filler, a wetting agent, a binder, a disintegrant or a lubricant.
5. The combined pharmaceutical composition for treating osimertinib-resistant non-small cell lung cancer according to claim 3, characterized in that: The pharmaceutical composition is in the form of any one or more of aqueous infusion, powder, lotion, tincture, oil, emulsion, ointment, plaster or aerosol.
6. Application of JK184 in combination with osimertinib in the preparation of drugs for the treatment of osimertinib-resistant non-small cell lung cancer.