CN115887461A - Application of EGFR inhibitor in preparation of medicine for treating spinal cord metastases - Google Patents

Abstract

The invention relates to application of a third-generation EGFR inhibitor in preparation of a medicine for treating spinal cord metastases, and particularly provides application of a compound shown in a formula I or a pharmaceutically acceptable salt thereof in preparation of a medicine for treating non-small cell lung cancer spinal cord metastases. The compound of the formula I or the pharmaceutically acceptable salt thereof has good curative effect on non-small cell lung cancer spinal cord metastases and has good clinical application prospect.

Description

Application of EGFR inhibitor in preparation of medicine for treating spinal cord metastases

Technical Field

The invention relates to the field of clinical medicine and pharmacy; in particular to application of a third-generation EGFR inhibitor in preparing a medicine for treating spinal cord metastases.

Background

Spinal cord tumors (spinal cord neoplasms) refer to neoplasms that occur in various structures of the spinal cord and spinal canal, resulting in various degrees of loss of spinal nerve function. Spinal cord tumors can be classified as primary and secondary spinal cord tumors. The primary spinal cord tumor refers to tumor occurring in adjacent tissues such as spinal cord and spinal canal nerves, blood vessels, meninges, adipose tissue, bony structures and embryonic residual tissue, and accounts for about 4-5% of the primary central nervous system tumor, and 56% of the primary central nervous system tumor is benign tumor. The secondary spinal cord tumor can be transferred from other systemic malignant tumors or malignant tumors in other parts of the central nervous system through blood, cerebrospinal fluid and other routes, and the tumors are not rare. Secondary tumors account for approximately 85% of spinal cord injuries caused by tumors.

At present, although there are some drugs available in clinical practice for treating spinal cord tumor, due to limited therapeutic effect and poor prognosis, there is still an urgent need to research related drugs, which can effectively treat spinal cord tumor and improve quality of life and survival time of patients with spinal metastasis.

The compound of the formula I is a small-molecule antitumor compound with independent intellectual property rights in Howson, and the methanesulfonic acid acitinib is the methanesulfonic acid salt of the compound of the formula I, can target EGFR to play antitumor activity, and belongs to EGFR-TKI:

the therapeutic effect of the compound of formula I on the treatment of advanced non-small cell lung cancer is well established, but the therapeutic effect in spinal metastases has not been studied.

Disclosure of Invention

The invention aims to provide more and more effective therapeutic drugs for treating the spinal cord metastases by researching the therapeutic effect of the compound shown in the formula I or the pharmaceutically acceptable salt thereof on the spinal cord metastases, particularly the spinal cord metastases caused by non-small cell lung cancer, and has wide clinical application prospects.

In one aspect, the present invention provides a use of a compound of formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for treating a myeloma,

wherein the pharmaceutically acceptable salt is selected from one or a combination of the following: mesylate, fumarate, maleate, acetate, hydrochloride, phosphate or sulfate.

In a preferred embodiment of the invention, the spinal myeloma is a neurogenic tumor, a meningioma, a neuroepithelial tissue tumor, an embryonic residual tissue tumor, a vascular tumor, or a metastatic tumor; metastases are preferred.

In a further preferred embodiment of the invention, the myelomas are derived from non-small cell lung cancer.

In a further preferred embodiment of the invention, the non-small cell lung cancer is locally advanced or metastatic lung cancer; preferably, the non-small cell lung cancer is bone metastases.

In a further preferred embodiment of the invention, the non-small cell lung cancer is caused by EGFR mutation, preferably one or more of exon 19 deletion, L858R mutation, T790M mutation, G719X mutation, S768I mutation or L861Q mutation.

In a further preferred embodiment of the invention, the unit dose of a compound of formula I or a pharmaceutically acceptable salt thereof is selected from 50mg to 300mg, preferably 55mg to 260mg, more preferably 55mg, 110mg, 220mg or 260mg, further preferably 55mg or 110mg.

In some embodiments, the method of treatment administers a composition to a patient at a frequency selected from the group consisting of: unit doses of 3 times a month, 4 times a month, 5 times a month, 2 times a month, 1 time a week, 2 times a week, 3 times a week, 4 times a week, 5 times a week, 6 times a week, 7 times a week, 1 time a day, 1 time two days, 2 times a day, 3 times a day.

In a further preferred embodiment of the invention, the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily at a daily dose of 110mg to 220mg per day, preferably 165mg per day.

In another aspect, the present invention provides a composition of a compound of formula I or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, diluent or excipient, preferably, the composition comprises lactose, microcrystalline cellulose, sodium carboxymethyl starch, sodium stearyl fumarate or magnesium stearate.

In a further preferred embodiment of the invention, the compounds of the formula I or their pharmaceutically acceptable salts are suitable for oral administration.

A unit dose (e.g., without limitation, a daily unit dose) of a compound of formula I or a pharmaceutically acceptable salt thereof is 50mg to 300mg (inclusive of ± 10%) of the compound of formula I, such as, without limitation, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 310, 320, 330mg, and values between any two of the foregoing values (not expressly listed but considered to be stated); more specifically, 55mg, 110mg, 220mg or 260mg may be used. Note: the dosages described in the formula are all based on the mesylate salt of the compound of formula I. It can be prepared in unit dosage form in a daily dose, administered once daily, and should be avoided from eating within 1 hour before administration to 2 hours after administration.

In a further preferred embodiment of the invention, the compound of formula I or a pharmaceutically acceptable salt thereof is administered alone, i.e. without the need to use it in combination with other drugs having an anti-tumor effect, but without excluding the use of some adjunctive drugs which do not have an anti-tumor effect.

In a further preferred embodiment of the present invention, the compound of formula I or a pharmaceutically acceptable salt thereof can be used in combination with other drugs for treating non-small cell lung cancer or bone marrow metastases, wherein the other drugs for treating non-small cell lung cancer or bone marrow metastases include cisplatin, carboplatin, pemetrexed, doxycycline, erlotinib, gefitinib, erlotinib, lapatinib, afatinib, or dacatinib.

In some embodiments, the route of administration is selected from: intramuscular, intraperitoneal, intravenous, subcutaneous, transdermal, intradermal, intranasal, intraocular, oral, sublingual, intratumoral, peritumoral.

Drawings

FIG. 1 is a scheme of animal experimental design.

Figure 2 is spinal cord and intracranial BLI signals in mice after treatment with amitinib.

Figure 3 is the fluorescent signal in the spinal cord of mice after treatment with amitinib.

Figure 4 is mouse intracranial fluorescence signal following treatment with amitinib.

Figure 5 is the change in body weight of mice after treatment with amitinib.

Figure 6 is a graph of survival of mice after treatment with amitinib.

FIG. 7 is the results of H & E and Ki-67 staining of mouse spinal cord tissue after treatment with Amitinib.

Detailed Description

Embodiments of the present application will be described in detail below with reference to specific examples. The following examples are merely illustrative of the present application and should not be construed as limiting the scope of the present application.

1. Animal testing

1. Purpose of the experiment:

the in vivo efficacy experiment proves the inhibition effect of the compound on the non-small cell lung cancer spinal cord metastasis.

2. Establishment of non-small cell lung cancer spinal cord metastasis mouse model

A spinal cord metastasis model is established by intracranial in-situ injection of tumor cells. The tumor seeding process is carried out in a super clean bench and is aseptic operation. The tumor cells used in the experiment are non-small cell lung cancer PC9-Luc cells transfected with luciferase, and the concentration of cell suspension during tumor seeding is 1.3X 108/ml. After intraperitoneal injection of pentobarbital sodium (50 mg/kg), the skin of the cranial vertex of the mouse is cut from two ears to two eyes along the center for about 1-2cm, and the skull is repeatedly wiped with a cotton swab to remove the fascia and expose cranial sutures. Punching at the position 1-1.5mm below the coronal suture of the skull and 2-2.5mm below the sagittal suture, keeping the skull of the mouse horizontal during punching, vertically inserting a cranial drill, slowly drilling at a low speed to avoid damaging brain tissues, and generally punching the right skull of the mouse. The mouse is placed on a brain stereotaxic apparatus, the needle point of the syringe is aligned with the hole on the skull, the needle insertion depth is 3.5mm away from the skull, the needle return is 0.5mm, 3 mu L of cell suspension (namely 4 multiplied by 105 cells) is injected, and the injection speed is 1 mu L/min. After the cell injection, the needle is withdrawn after the cell injection is finished for 1-2 minutes, and the wound is pressed by a cotton swab. The skin wound was adhered using medical adhesive. The mice are placed on a heating blanket, and after the mice are recovered, the mice are placed back into an animal room for continuous breeding.

3. Test protocol

In a mouse model of spinal cord metastasis, when the fluorescence signal in the spinal column of the mouse reaches 2X 10 ⁷ The effect of long-term administration of amitinib (once a day) was observed at photon/second (about 2-3 weeks). Mice were divided into three groups-control, almetinib 5mg/kg and Almetinib 25 mg/kg. BLI signals were measured once a week and the body weight and survival of the mice over two months were recorded. H mouse spinal cord tissue&E and Ki67 immunohistochemistry were used to observe cell proliferation status. The specific experimental protocol design is shown in figure 1.

H & E staining:

animal spinal cord tissue was fixed in 4% paraformaldehyde solution and dehydrated with different concentrations of xylene and ethanol. Rinsed with distilled water and stained with hematoxylin stain for 15 minutes. Next, alcohol differentiation with hydrochloric acid, eosin staining, alcohol dehydration, xylene remained transparent. The final step of installation is accomplished by balm neutrality. All processes must work with the vent. All sections were observed and captured at 400 x using an olympus BX51 microscope (Tokyo, japan).

Immunohistochemistry:

frozen spinal cord tissue was obtained in a thickness of 10 μm using a cryomicrotome (CM 1900, leica, germany) and sectioned. Subsequently, after deparaffinization, antigen retrieval and removal of endogenous peroxidase steps, a primary antibody solution of Ki67 (1. All sections were observed and captured at 400 x using an olympus BX51 microscope (Tokyo, japan).

4. Results of the experiment

(1) Intracranial and intraspinal BLI signaling in mice

As can be seen from the results of fig. 2, the BLI signals in the spinal cord and the intracranial space of the control mice were very strong, but the BLI signals in the spinal cord and the intracranial space of the mice in the armenitinib group were significantly reduced and dose-dependent. Experimental results show that the amitinib can effectively inhibit spinal cord metastasis and intracranial metastasis of tumor cells in a mouse body.

(2) Intracranial and intraspinal immunofluorescence signals in mice

After 16 days of modeling, growth and metastasis of PC9-Luc cells were assessed weekly using bioluminescence imaging. The bioluminescent signal in spinal cord was significantly reduced in mice of the 25mg/kg group of Almetinib as compared to the control group, indicating that growth and metastasis of spinal cord and intracranial tumor cells were inhibited. As can be seen from Table 1 and FIG. 3, the fluorescence signal in the spinal cord of the control mice was significantly increased, the bioluminescent signal in the spinal cord of the mice in the group of 5mg/kg of Almetinib was not significantly changed, and the bioluminescent signal in the spinal cord of the mice in the group of 25mg/kg of Almetinib was significantly decreased. The result shows that the amitinib can effectively inhibit the proliferation of tumor cells in the spinal cord. As can be seen from Table 2 and FIG. 4, the intracranial fluorescence signals of the control mice significantly increased, the intracranial bioluminescence signals of the 5mg/kg Almerinib mice did not significantly change, and the intracranial bioluminescence signals of the 25mg/kg Almerinib mice significantly decreased. The result shows that the amitinib can effectively inhibit the proliferation of tumor cells in the intracranial.

TABLE 1 bioluminescent signals in the spinal cord of mice

Note: ND means animal death was not detected.

TABLE 2 intracranial bioluminescent signals in mice

Note: ND means animal death was not detected.

(3) Survival of mice after amitinib treatment

As can be seen from the results in fig. 5, the body weight of the mice did not significantly decrease after administration of amitinib, relative to the control group. As can be seen from the results of fig. 6 and table 3, the survival time of nude mice in the group administered with amitinib was longer than that of the control group.

Table 3 percent survival of spinal cord transferred mice%

(4) Mouse spinal cord H & E and Ki-67 staining results

As can be seen from the results in fig. 7, after treatment, spinal cords of high dose ametinib and control mice were dissected for H & E and Ki-67 staining. The H & E results showed a significant reduction in tumor area in the group treated with Almetinib, and the Ki-67 immunohistochemistry results showed a significant inhibition of tumor cell proliferation in the high dose Almetinib group compared to the control group (control group: (19.3. + -. 3.07)%; almetinib-treated group: (55.35 ±. 7.07)%). The result shows that the amitinib can effectively inhibit NSCLC spinal cord metastasis in vivo.

Claims (10)

1. The application of the compound of the formula I or the medicinal salt thereof in preparing the medicament for treating the spinal cord tumor,

wherein the pharmaceutically acceptable salt is selected from one or a combination of the following: mesylate, fumarate, maleate, acetate, hydrochloride, phosphate or sulfate.

2. The use according to claim 1, wherein the spinal myeloma is a neurogenic tumor, a meningioma, a neuroepithelial tissue tumor, an embryonic residual tissue tumor, a vascular tumor, or a metastatic tumor; metastases are preferred.

3. The use of claim 2, wherein the spinal cord is derived from non-small cell lung cancer.

4. The use of claim 3, wherein the non-small cell lung cancer is locally advanced or metastatic lung cancer; preferably, the non-small cell lung cancer is bone metastases.

5. Use according to claim 3, wherein the non-small cell lung cancer is caused by an EGFR mutation, preferably one or more of an exon 19 deletion, an L858R mutation, a T790M mutation, a G719X mutation, an S768I mutation or an L861Q mutation.

6. Use according to claim 3, wherein the non-small cell lung cancer is a non-squamous cell carcinoma, preferably an adenocarcinoma, more preferably a stage II or III adenocarcinoma; optionally, the locally advanced or metastatic non-small cell lung cancer develops brain metastases.

7. Use according to claim 1, wherein the unit dose of the compound of formula I or a pharmaceutically acceptable salt thereof is selected from 50mg to 300mg, preferably 55mg to 260mg, more preferably 55mg, 110mg, 220mg or 260mg, further preferably 55mg or 110mg.

8. Use according to claim 1, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is administered once daily at a daily dose of 110mg to 220mg per day, preferably 165mg per day.

9. Use according to claim 1, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is a composition further comprising a pharmaceutically acceptable carrier, diluent or excipient, preferably wherein the composition comprises lactose, microcrystalline cellulose, sodium carboxymethyl starch, sodium stearyl fumarate or magnesium stearate.

10. Use according to any one of claims 7 to 9, wherein the compound of formula I or a pharmaceutically acceptable salt thereof is for oral administration or injection.

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