Application of CDK4 / 6 inhibitor in preparation of medicine for preventing or treating ototoxicity caused by platinum chemotherapy medicine and radiotherapy

Abstract

The invention discloses application of a CDK4 / 6 inhibitor in preparation of medicines for preventing or treating ototoxicity caused by platinum chemotherapy and radiotherapy. Through combined use of the CDK4 / 6 inhibitor and anti-cancer treatment, cochlea hair cell loss can be significantly reduced, a hearing threshold is reduced, and the anti-tumor effect of cis-platinum / radiotherapy is not affected. The device is especially suitable for breast cancer, head and neck cancer and other tumor patients needing long-term use of platinum drugs or radiotherapy.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a new use of CDK4 / 6 inhibitors in preventing or treating ototoxicity caused by platinum-based chemotherapeutic drugs and radiotherapy. Background Art

[0002] Platinum-based chemotherapeutic drugs include cisplatin, carboplatin, nedaplatin, cycloplatin, oxaliplatin, and lobaplatin, etc., which are commonly used first-line tumor chemotherapeutic drugs in clinical practice. Although such chemotherapeutic drugs have strong killing ability against tumor cells, the extremely strong ototoxic side effects of such drugs not only limit their further extensive and long-term use in clinical practice, but also significantly reduce the quality of life of patients. Research shows that among patients receiving cisplatin chemotherapy, about 30-40% of adult patients and up to 70% of pediatric patients will develop irreversible hearing loss (van AS JW, Cochrane Database Syst Rev, 2016). In addition, radiotherapy is the main treatment method for head and neck tumors. However, the hearing of patients with head and neck tumors receiving radiotherapy is significantly reduced (Wang J, Translational Oncology, 2015). Therefore, developing drugs that can prevent hearing damage after platinum-based drug chemotherapy and radiotherapy has broad clinical application prospects. Summary of the Invention

[0003] The present invention provides a use of a CDK4 / 6 inhibitor in the preparation of a drug for preventing or treating ototoxicity caused by platinum-based chemotherapeutic drugs and radiotherapy, wherein the CDK4 / 6 inhibitor is used in combination with a platinum-based chemotherapeutic drug or radiotherapy.

[0004] Preferably, the CDK4 / 6 inhibitor is selected from at least one of palbociclib, ribociclib, and abemaciclib.

[0005] Preferably, the platinum-based chemotherapeutic drug is selected from at least one of cisplatin, carboplatin, nedaplatin, oxaliplatin, and lobaplatin.

[0006] Preferably, the radiotherapy includes at least one of conventional external irradiation, intracavitary afterloading radiotherapy for nasopharynx, or stereotactic radiotherapy.

[0007] Preferably, the ototoxicity is manifested as cochlear hair cell loss and / or an increase in hearing threshold by 20-50 dB.

[0008] Preferably, the CDK4 / 6 inhibitor is used 0-8 hours before radiotherapy or administration of a platinum-based chemotherapeutic drug.

[0009] Preferably, the tumors targeted by the use include at least one of breast cancer, lung cancer, and head and neck tumors.

[0010] Preferably, the dosage of the CDK4 / 6 inhibitor is 35 - 75 mg / kg / day, and the dosage of the platinum-based chemotherapeutic drug is 4 - 32 mg / kg / week.

[0011] Preferably, the drug is an oral preparation or an injection, and is used simultaneously or sequentially with a platinum-based chemotherapeutic drug or radiotherapy.

[0012] Meanwhile, the present invention also provides a tumor treatment drug composition, comprising a therapeutically effective amount of a CDK4 / 6 inhibitor, a platinum-based chemotherapeutic drug, and / or a radiotherapy adjuvant (mass of CDK4 / 6 inhibitor: mass of platinum-based chemotherapeutic drug 35 - 75 mg: 4 - 32 mg), and is used for synergistically inhibiting tumor growth and reducing ototoxic side effects.

[0013] Preferably, the radiotherapy rays are α, β, γ rays emitted by radioactive isotopes; X-rays with different energies generated by X-ray therapy machines and various accelerators; electron beams, fast neutrons, proton beams, negative pion beams, and other heavy particle beams generated by various accelerators, etc., but not limited thereto.

[0014] Preferably, the tumor is one or more of breast cancer, lung cancer, liver cancer, colorectal cancer, gastric cancer, esophageal cancer, intracranial tumor, nasopharyngeal tumor, oral and maxillofacial tumor, etc., but not limited thereto.

[0015] The present invention first discovers that the CDK4 / 6 inhibitor can prevent cochlear hair cell damage and hearing loss caused by platinum-based chemotherapy and radiotherapy by inhibiting terminally differentiated hair cells from entering the cell cycle. phase cycle, thus reducing the apoptosis of hair cells to The inhibitor, when used in combination with anti-cancer treatment, can not only maintain the tumor killing effect but also protect the auditory system by inhibiting the CDK4 / 6 pathway. Animal experiments show that the combined treatment regimen increases the survival rate of cochlear hair cells in cisplatin-treated mice by 30% - 60%, reduces the hearing threshold by 20 - 40 dB, and the tumor inhibitory effect is equivalent to that of cisplatin alone. Description of the Drawings

[0016] Figure 1 It is a diagram showing the survival status of cochlear hair cells and changes in hearing function of mice in the cisplatin group / cisplatin + CDK4 / 6 inhibitor group.

[0017] Figure 2 It is a diagram showing the change in tumor volume of mice in the cisplatin group / cisplatin + CDK4 / 6 inhibitor group.

[0018] Figure 3 It is a diagram showing the survival status of cochlear hair cells of mice in the radiotherapy group / radiotherapy + CDK4 / 6 inhibitor group.

[0019] Figure 4 In a - d shows the anti-apoptotic effect of the CDK4 inhibitor on the hearing hair cells of mice. Detailed implementation mode

[0020] The following further describes in detail the preferred embodiments of the present invention in conjunction with the accompanying drawings: The following embodiments facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following embodiments are all conventional methods unless otherwise specified. The test materials used in the following embodiments are all purchased from conventional biochemical reagent companies unless otherwise specified.

[0021] Hearing test method: In this experiment, auditory brainstem response (ABR) was used to conduct pure-tone hearing tests on mice. After the mice were anesthetized, a signal electrode, a reference electrode, and a ground electrode were inserted. Through the TDT test system, sound waves of 4 kHz - 32 kHz and 20 dB - 90 dB were played to determine the responses of the mice to sound waves of different frequencies and intensities, thereby obtaining the hearing thresholds of each frequency band of the mice. Hair cell staining technique: In this experiment, after the hearing tests of the mice were completed, the mice were euthanized, the cochleas were taken out, fixed with 4% paraformaldehyde at 4 °C for 24 hours, then decalcified with 10% EDTA, the basilar membranes were dissected out, and the basilar membranes were stained with the primary antibody (1:400) overnight. After washing with PBS, they were stained with the secondary antibody (1:200), and stained with phalloidin (a hair cell marker) (1:400) for one hour, followed by DAPI staining. Subsequently, the samples were placed on slides and sealed, and observed under a confocal microscope.

[0022] Tumor transplantation: Inject 1X10 6 breast cancer tumor cells into the mammary fat pad of mice. Two weeks later, when the tumor volume of the mice reaches 150 - 250 mm 3 , start the drug administration experiment. Measure the length and width of the tumor daily, and calculate the change in tumor volume according to the formula volume = length × width × width / 2.

[0023] Example 1: Detect the effects of cisplatin and CDK4 inhibitor on cochlear hair cells and hearing in Balb / c mice Wild-type Balb / c (female, 4 weeks) mice were divided into the following three groups and given drugs respectively: saline, cisplatin, CDK4 inhibitor, cisplatin + CDK4 inhibitor (5 mice in each group). Cisplatin (16 mg / kg) was given on the 1st day of the experiment, and the CDK4 inhibitor (75 mg / kg, and the CDK4 inhibitor was given 4 hours before cisplatin on the 1st day) was given on the 1st, 3rd, and 5th days of the experiment. Pure-tone hearing tests were performed before drug administration and 14 days after drug administration. At the same time, after 14 days of drug administration, the cochleas of the mice were taken for immunofluorescence staining to observe the loss of hair cells in each group of mice.

[0024] Example 2: Detect the effects of cisplatin and CDK4 inhibitor on tumors in mice Wild-type Balb / c mice were transplanted with triple-negative breast cancer cells in the mammary fat pads. After successful tumor modeling, the tumor-bearing mice were divided into three groups for treatment as described above. After 28 days of drug administration, the tumors of the mice were sampled, and the therapeutic effect of the drug on the tumors was evaluated by comparing the tumor sizes. The CDK4 inhibitors used in the present invention are all marketed drugs, which have passed clinical phase I, II, and III tests and have been widely used to treat estrogen receptor-positive breast cancer for more than 10 years in many countries around the world, including China and the United States. They have been proven to be safe.

[0025] Example 3: Detection of the effects of radiotherapy and CDK4 inhibitors on cochlear hair cells and hearing in mice Wild-type Balb / c mice were divided into the following three groups for separate treatment: non-radiotherapy control, radiotherapy, and radiotherapy + CDK4 inhibitor. After 36 hours of radiotherapy, the cochleas of the mice were sampled for immunofluorescence staining to observe the loss of hair cells in each group of mice.

[0026] Figure 1 a-e in it show the chemoprotective effect of CDK4 inhibitors on cochlear hair cells in mice with hearing. Compared with the cisplatin group, there was almost no loss of cochlear hair cells in the cisplatin + CDK4 inhibitor group of mice, and at the same time, the hearing thresholds at each frequency increased less. This study shows that inhibiting CDK4 can effectively rescue cisplatin-induced cochlear hair cell and hearing loss.

[0027] Figure 1 d demonstrates that the combination of drugs can reduce more than 20 - 50% of hair cell damage. The protective effect in the high-frequency region (24k - 32k) suggests that CDK4 inhibitors may be more suitable for children (sensitive to high-frequency hearing) or patients with head and neck cancer radiotherapy (the cochlear high-frequency region is vulnerable to radiation damage).

[0028] Figure 1 e demonstrates that CDK4 inhibitors can selectively reduce high-frequency hearing loss caused by cisplatin chemotherapy (the protection efficiency in the range of 16k - 32kHz reaches 30 - 80%), providing a key experimental basis for the development of the "chemotherapy - ear protection" combination therapy.

[0029] Figure 2 It shows that CDK4 inhibitors do not affect the therapeutic effect of cisplatin on tumors in mice. As can be seen from the figure, compared with the control group (saline group), the tumors of mice in the cisplatin group and the cisplatin + CDK4 inhibitor group were significantly smaller, and at the same time, there was no difference in the tumor sizes between the cisplatin + CDK4 inhibitor group of mice and the cisplatin group of mice.

[0030] Figure 3a-g show the protective effect of CDK4 inhibitor on the γ-ray radiotherapy of mouse cochlear hair cells. Compared with the significantly reduced cochlear hair cells in the radiotherapy group, there was almost no loss of cochlear hair cells in the mice of the radiotherapy + CDK4 inhibitor group. This study indicates that inhibiting CDK4 can effectively rescue the loss of cochlear hair cells caused by radiotherapy.

[0031] Figure 4 a-d show the anti-apoptotic effect of CDK4 inhibitor on mouse cochlear hair cells. On the 7th / 14th day after drug administration, compared with the cisplatin group, the number of p53-positive hair cells in the cisplatin + cdk4 inhibitor group was significantly reduced. p53 is a marker of apoptosis, indicating that inhibiting CDK4 can effectively rescue the loss of cochlear hair cells caused by platinum drugs through anti-apoptosis.

[0032] The above content is a further detailed description of the present invention in combination with specific preferred embodiments. It cannot be determined that the specific implementation of the present invention is only limited to these descriptions. For those of ordinary skill in the technical field to which the present invention belongs, without departing from the concept of the present invention, several simple deductions or substitutions can be made, which should all be regarded as belonging to the protection scope of the present invention.

Claims

1. Use of a CDK4 / 6 inhibitor in the preparation of a drug for preventing or treating ototoxicity caused by platinum chemotherapy drugs and radiotherapy, characterized in that: The CDK4 / 6 inhibitor is used in combination with a platinum-based chemotherapeutic drug or radiotherapy.

2. The use according to claim 1, wherein: The CDK4 / 6 inhibitor is selected from at least one of palbociclib, ribociclib, and abemaciclib.

3. The use according to claim 1, characterized in that: The platinum-based chemotherapeutic drug is selected from at least one of cisplatin, carboplatin, nedaplatin, oxaliplatin, and lobaplatin.

4. The use according to claim 1, characterized in that: The radiotherapy includes at least one of conventional external beam radiotherapy, intracavitary brachytherapy for nasopharynx, or stereotactic radiotherapy.

5. The use according to claim 1, characterized in that: The ototoxicity is manifested as cochlear hair cell loss and / or a 20 - 50 dB increase in hearing threshold.

6. The use according to claim 1, characterized in that: The CDK4 / 6 inhibitor is used 0 - 8 hours before radiotherapy or administration of the platinum-based chemotherapeutic drug.

7. The use according to claim 1, characterized in that: The tumors targeted by the use include at least one of breast cancer, lung cancer, and head and neck tumors.

8. The use according to claim 1, characterized in that: The administration dose of the CDK4 / 6 inhibitor is 35 - 75 mg / kg / day, and the administration dose of the platinum-based chemotherapeutic drug is 4 - 32 mg / kg / week.

9. The use according to claim 1, wherein: The drug is an oral preparation or an injection, and is used synchronously or sequentially with the platinum-based chemotherapeutic drug or radiotherapy.

10. A tumor treatment pharmaceutical composition, characterized in that: It contains a therapeutically effective amount of a CDK4 / 6 inhibitor, a platinum-based chemotherapeutic drug, and / or a radiotherapy adjuvant, and is used to synergistically inhibit tumor growth and reduce ototoxic side effects.

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