WO2017129094A1 - Use of egfr/her2 receptor tyrosine kinase inhibitor in preparing drugs for treating cancers induced by her2 mutation - Google Patents

Abstract

The present invention relates to the use of an EGFR/HER2 receptor tyrosine kinase inhibitor in preparing drugs for treating cancers induced by HER2 mutation. In particular, the present invention relates to the use of a compound as shown in formula A or a pharmaceutically acceptable salt thereof in preparing drugs for treating cancers induced by HER2 mutation.

Description

Use of an EGFR/HER2 receptor tyrosine kinase inhibitor for preparing a medicament for treating HER2 mutation cancer Technical field

Use of an EGFR/HER2 receptor tyrosine kinase inhibitor for the preparation of a medicament for treating a HER2 mutant cancer.

Background technique

Lung cancer has become the leading cause of cancer deaths worldwide, both male and female. According to the statistics of the 2015 China Cancer Registration Annual Report, the incidence and mortality of lung cancer in China both occupy the first place. The incidence and mortality of lung cancer increase with age. Generally, the incidence of lung cancer increases significantly after the age of 40, reaching a peak at around 75 years old, and then decreases (Shi Yuankai, Sun Yan. Handbook of Clinical Oncology. Beijing: People Health Press, 2015: 315-341). In lung cancer, non-small cell lung cancer (NSCLC) accounts for approximately 85% of all lung cancer patients (Siegel R, Ma J, Zou Z, et al. Cancer statistics. CA Cancer J Clin, 2014, 64(1): 9- 29). Adenocarcinoma is the most common pathological type in non-small cell lung cancer in developed countries, accounting for about 40%. Most NSCLCs are locally advanced or distantly transferred and cannot be surgically removed.

The first line of treatment for metastatic NSCLC depends on the type of pathology and genetic changes. EGFR-TKI is recommended for patients with positive EGFR mutations, and crizotinib is recommended for ALK-positive patients (Shi Yuankai, Sun Yan. Handbook of Clinical Oncology. Beijing: People's Medical Publishing House, 2015: 315-341). Non-squamous cell carcinoma patients with negative gene expression are recommended to receive pemetrexed or other platinum-containing combination chemotherapy, and combined with endostatin (YH-16) or cetuximab (Cetuximab). For patients with first-line treatment to achieve disease control (complete remission, partial remission, and stabilization), maintenance therapy is optional. Currently, evidence-based medical evidence supports pemetrexed (non-squamous cell carcinoma) and gemcitabine. For patients with EGFR mutations, epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) can be maintained. For second-line treatment, docetaxel, pemetrexed, and EGFR-TKI are available. There is no clear recommended treatment for third-line treatment, and a large number of clinical trials are exploring other effective treatment methods (Shi Yuankai, Sun Yan. Handbook of Clinical Oncology [M]. Beijing: People's Medical Publishing House, 2015: 315-341) . In addition to the above mentioned targets, lung cancer studies have also revealed a number of genetic abnormalities, including amplification of MET and FGFR1, PIK3CA, AKT, KRAS, NRAS, BRAF, MEK1, AKT1, FGFR2, DDR2 and HER2 mutations, And RET and ROS1 rearrangement, etc. (Mazières J, Peters S, Lepage B, et al. Lung cancer that harbors an HER2 mutation: Epidemiologic characteristics and therapeutic perspectives. J Clin Oncol, 2013, 31(16): 1997-2003).

The human epidermal factor receptor 2 (HER2) gene belongs to the HER tyrosine receptor family, and the HER2 mutation mainly occurs in exon 20, accounting for 2-4% of NSCLC patients (Clin Cancer Res. 2012, 18 : 4910-4918; Cancer Res. 2005, 65: 1642-1646; Lung Cancer. 2011, 74: 139-144.). However, in 2013, Barlesi et al found that 10,000 patients with advanced NSCLC were found in a large molecular marker study. The HER2 mutation rate was 0.9% (J Clin Oncol, 2013, 31 (suppl): abstr 8000). Mazières et al reported that 65 (1.7%) HER2 mutations were detected in 3800 lung adenocarcinoma patients (J Clin Oncol, 2013, 31(16): 1997-2003). HER2 mutations are mutually exclusive with other driver gene abnormalities (EGFR, KRAS, BRAF mutations, and ALK rearrangements, etc.) (Lung Cancer, 2015, 87(1): 14-22). Suzuki et al (Lung Cancer, 2015, 87(1): 14-22) also reported that HER2 mutation is an independent prognostic factor for poor prognosis in patients with invasive non-small cell lung adenocarcinoma (p=0.005). In clinical trials, Tomizawa et al observed that a patient with lung-adenostatic HER2-mutant lung adenocarcinoma who was treated with platinum-based chemotherapy and gefitinib was effective for trastuzumab plus vinorelbine (as a third-line treatment). (Lung Cancer, 2011, 7: 139-144). Another study of the clinical features and efficacy of Mazières et al in patients with HER2 mutations in NSCLC showed that a total of 16 patients with HER2 mutations in non-small cell lung adenocarcinoma received HER2 targeted drug therapy, some of whom received 2 (3 cases) or 4 (1 case) different HER2 targeting drugs (J Clin Oncol, 2013, 31(16): 1997-2003). A total of 16 patients with HER2 mutations in non-small cell lung adenocarcinoma received 22 HER2 targeted drug treatments with measurable outcomes, including 4 disease progression (PD) and 7 disease stabilization (SD, 32%). ), 11 patients had partial remission (PR, 50%), and the disease control rate (DCR) was 82%. Of these, 15 patients treated with trastuzumab had a DCR of 96%, 4 patients with afatinib had a DCR of 100%, and 2 patients had lapatinib and 1 The efficacy of patients using masatinib was evaluated as PD. The median PFS of 15 patients who used HER2 targeted drugs on the first line was 5.1 months (Mazières J et al, J Clin Oncol, 2013, 31(16): 1997-2003). Another clinical trial of natalin monotherapy or temsirolimus in the treatment of HER2 mutant non-small cell lung cancer (NCT01827267), published in September 2014, showed that 13 patients took neratinib monotherapy Among the subjects, PR 0 cases, SD accounted for 54%, SD ≥ 12 weeks accounted for 31%; that is, ORR was 0, DCR was 54%. Of the 14 patients taking neratinib plus temsirolimus, 3 patients (21%) had PR, 11 patients (79%) had SD, and 12 patients (64%) had SD ≥ 12 weeks; that is, ORR was 21%, DCR It is 100%. The PFS of neratinib alone or in combination with temsirolimus was 2.9 months and 4 months, respectively (http://www.bioportfolio.com/resources/trial/131948/Neratinib-With-and-Without -Tems irolimus-for-Patients-With-HER2-Activating-Mutations-in.html). The above multiple test results show that some HER2 and / or EGFR targeted drugs may have anti-tumor efficacy in HER2 mutant non-small cell lung cancer patients, while some HER2 and / or EGFR targeted drugs are ineffective, as for a specific Whether HER2 and/or EGFR targeted drugs are effective for HER2-mutated cancers is difficult to predict.

CN102471312B discloses a compound represented by the following formula A (chemical name (E)-N-[4-[[3-chloro-4-(2-pyridylmethoxy)phenyl]amino]-3-cyano- 7-Ethoxy-6-quinolinyl]-3-[(2R)-1-methylpyrrolidin-2-yl]prop-2-enamide), and disclosed that it has strong EGFR and HER2 Inhibition and anticipation of its possible use in the treatment of cancers overexpressing EGFR and HER2,

CN102933574B discloses a series of pharmaceutically acceptable salts of the compounds of formula A. CN103974949B discloses crystalline forms of dimaleate salts of the compounds of formula A.

However, none of the above publications disclose the effect of the compound of formula A on the treatment of HER2-mutant cancer.

Summary of the invention

The present inventors have surprisingly found that Compound A or a pharmaceutically acceptable salt thereof has an amazing effect on the treatment of HER2 mutant cancer, and thus completed the present invention.

In a preferred embodiment of the invention, the cancer is a lung cancer, a breast cancer or a gastrointestinal cancer. Preferably, the lung cancer is non-small cell lung cancer, and further preferably a HER2 mutant non-small cell lung adenocarcinoma, particularly a late stage non-small cell lung adenocarcinoma patient with a HER2 mutation. Preferably, the gastrointestinal cancer is gastric cancer.

In the present invention, the HER2-mutated cancer refers to a cancer driver mutation in which a HER2 gene can be detected in these cancer patients, and the type of the mutation is mainly a non-frameshift insertion mutation of exon 20, Followed by some missense mutations and small insertion and deletion mutations in the tyrosine kinase domain and other segments, such as 772-775 YVMA duplication (the amino acid at position 772-775 in the original protein is a YVMA, due to the insertion of 12 genes in the gene The base forms another pair of YVMA, causing a repetition of YVMA), G776delinsVC (the amino acid G of 776 becomes V due to the insertion of 3 bases at the corresponding site, and the amino acid C is inserted, which can also be expressed as insG776V C Or G776>VC), P780_Y781insGSP (the insertion of GSP three amino acids between amino acids 780-781 due to the insertion of 9 bases at the corresponding site), V777L (the amino acid V of 777 due to a missense mutation of one base becomes L), L755S, S310F, S310Y, G309A, G309E, D769Y, D769H, V842I, L866M, R896C, L755_E757delinsS (3 amino acids 755-757 are treated with an ammonia due to deletion of 6 bases at the corresponding site) Substituent S substitution), L869R, L841V, G776V, L755-T759Del (amino acid deletion 755-759 due to deletion of 15 bases at the corresponding site), G776L, V777L, L755P, and S779_P780insVGS (VESS three amino acids inserted between amino acids 779-780 due to insertion of 9 bases at the corresponding site) (amino acid sequence number refers to P04626ERBB2_HUMAN).

In a preferred embodiment of the invention, the cancer is a cancer that is negative for EGFR mutation or negative for ALK fusion gene, or both are negative.

In a preferred embodiment of the invention, the cancer is a cancer that progresses after chemotherapy, radiation therapy or targeted therapy. That is, the patient with the cancer is not controlled after chemotherapy, radiotherapy or targeted therapy, and continues to progress. The chemotherapy described therein may be treated with various conventional chemotherapeutic drugs, such as alkylating agents (eg, cyclophosphamide, ifosfamide, melphalan, busulfan, nimestin, ramustine, da Carbazine, temozolomide, nitrogen mustard, dibromomannitol, etc.), platinum complexing agents (eg cisplatin, carboplatin, oxaliplatin, etc.), metabolic antagonists (eg methotrexate, 5-fluorouracil, Fluoride, gemcitabine, capecitabine, fulvestrant, pemetrexed, etc.), plant alkaloids (eg vincristine, vinblastine, vindesine, etoposide, docetaxel, paclitaxel, y Liticon, vinorelbine, mitoxantrone, vinorelbine, topotecan, etc.), hormone anticancer agents (such as leuprolide, goserelin, dutasteride, dexamethasone, tamoxifen Fen et al), proteasome inhibitors (eg, bortezomib, lenalidomide, etc.), aromatase inhibitors (eg, exemestane, letrozole, anastrozole, etc.), preferably selected from carboplatin Chemotherapy is performed in one or more of cisplatin, oxaliplatin, pemetrexed, gemcitabine or docetaxel. The targeted therapy can be treatment with one or more selected from the group consisting of an EGFR inhibitor and a VEGFR inhibitor. These targeted drugs are well known in the art, for example, the EGFR inhibitor may be selected from one or more of gefitinib, erlotinib, ectinib and afatinib; the VEGFR inhibitor is selected from the group consisting of One or more of nitinib, apatinib, and faritinib.

In the present invention, the compound A is preferably in the form of a pharmaceutically acceptable salt thereof, particularly a maleate or a dimaleate, in practical use.

In the present invention, the daily dose of Compound A or a pharmaceutically acceptable salt thereof may range from 1 mg/kg to 20 mg/kg, preferably from 2 mg/kg to 10 mg/kg, more preferably from 4 to 8 mg/kg. . For adults, it is preferably 100 mg to 1000 mg, preferably 240 to 560 mg, more preferably 320 to 480 mg, based on the compound A. Among them, for Asians, the daily dosage can range from 240 to 400 mg, especially 400 mg.

Compound A or a pharmaceutically acceptable salt thereof may also be formulated with a pharmaceutically acceptable carrier in the form of a composition well known in the art, such as tablets, capsules, granules, injections and the like. The invention also relates to the use of a pharmaceutical composition comprising Compound A for a HER2 mutated cancer.

DRAWINGS

Figure 1 shows the inhibition of the proliferation of tumor cells in vitro by Compound A and lapatinib (a dose-response curve).

detailed description

The invention is further described in the following examples, but these examples are not intended to limit the scope of the invention.

Example 1: Effect of Compound A and Lapatinib on Proliferation of ATCC H1781 Cells Cultured in Vitro.

1. Test drug

Drug name: dimaleate salt of Compound A (batch number S0915100514), lapatinib di-p-toluenesulfonate (batch number 20090105). Preparation method: all were prepared in DMSO.

2. Cell line

ATCC H1781 cells were obtained from Shanghai Pulmonary Hospital. The cells were HER2 mutation (InsG776V, C). The patient was a 66-year-old woman with gestational lung adenocarcinoma and was cultured in PRIM 1640 medium containing 10% fetal bovine serum (FBS).

3, reagents and instruments

PRIM 1640 was purchased from Gibco BRL; fetal bovine serum was purchased from Gibco; a multi-purpose microplate reader was purchased from BioTek; and Sulforhodamine B (SRB) was purchased from Sigma.

4. Test method

The inhibitory effect of drugs on the proliferation and growth of tumor cells was detected by SRB protein staining. The main steps are as follows:

The logarithmic growth phase cells were seeded in 96-well culture plates, and the corresponding concentrations of the drugs (1-10000 nM) were added, and the respective wells were set at the same concentration, and the corresponding concentration of the vehicle control was set. Tumor cells were cultured for 72 h at 37 ° C under 5% CO ₂ . The cells were stained with SRB at room temperature, finally dissolved in Tris solution, and the OD value was measured at a wavelength of 510 nm by a microplate reader (BioTek), and the cell growth inhibition rate was calculated by the following formula:

Inhibition rate = (OD value _{control hole-} OD value _{administration hole} ) / OD value _{control hole} × 100%

The half-inhibitory concentration IC _{50 was} calculated according to the nonlinear regression method according to the respective concentration inhibition rates.

5, test results

The results showed that Compound A had a good inhibitory effect on HER2 mutant lung adenocarcinoma cells, while lapatinib had a poor effect. The specific results are shown in Table 1 and Figure 1:

Table 1 Summary of Compound A and Lapatinib for In vitro Culture of Tumor Cells ATCC H1781 Value-Added IC ₅₀

drug	IC 50 (nM)	95% confidence interval	R 2
Compound A	59	38.92to 89.19	0.9805
Lapatinib	2430	1354to 4362	0.9593

R is the correlation coefficient.

Example 2: Effect of Compound A and Lapatinib on the Activity of HER2 Recombinant Protease in Vitro

1. Test drug

Compound A (batch number SHR120201-002-06), and lapatinib (batch number SHR115758-010-17) were all supplied by Jiangsu Hengrui Medicine, and staurosporin was purchased from MedChem (Monmouth Junction, NJ) (batch number MC). -2104).

2. Recombinant protein

Recombinant human protein HER2WT (Lot#W353-1) and 5 HER2 mutant proteins (A775_G776insYVMA:lot#Z1251-6; D769H:lot#K1683-5; D769Y:lot#P1688-9; V777_G778insCG:lot#Z1287-3; V777L:lot#K1850-3) are purchased from SignalChem (Richmond, BC V6V 2J2, CANADA). These recombinant proteins are the amino acid 676 amino acid to the C-terminal 1255 amino acid polypeptide, both from baculovirus in Sf9 insect cells. Expressed in it, and the N-terminus is labeled with GST. The EGFR gene index number is NM_004448. The protein purity of WT HER2 and the four HER2 muteins (A775_G776ins YVMA, D769H, D769Y, V777_G778insCG) was greater than 85%, and the other HER2 mutein V777L was greater than 90% pure.

3. Test method

Using bovine serum albumin (BSA) as a substrate, the reaction system (20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO) Add 10 μM-0.5 nM of the test compound (Compound A or the positive control staurosporine), and then add 33P-ATP 0.01 μCi/μl (Perkin Elmer) to initiate the phosphorylation reaction to determine the enzymatic activity of EGFR. . The results showed that Compound A had a strong inhibitory effect on the enzymatic activity of recombinant human wild type EGFR and exon 20 insertional mutation, and with the increasing concentration of compound A, the total enzyme activity of EGFR gradually decreased. A concentration dependent relationship is presented. Taking the logarithm of the concentration as the abscissa, the probability unit of the corresponding concentration inhibition rate is the ordinate, and the IC50 value is calculated by Prism4 software (GraphPad).

4 test results

The results showed that Compound A had a good inhibitory effect on HER2 mutant lung adenocarcinoma cells, of which lapatinib, neratinib and staurosporin were used as control compounds. The specific results are shown in Table 2:

Compound A ₅₀ Table 2 summarizes the in vitro activity of recombinant protease HER2 IC

Example 3: Effect of Compound A and Lapatinib on Proliferation of HER2 Mutant MCF10A Cell Line in Vitro.

1. Test drug

Drug name: Compound A (batch number S0915151219), lapatinib (batch number SHR115758-010-17). Preparation method: all were prepared in DMSO.

2. Cell line

MCF10A cells were purchased from ATCC, and the cells were used as mother cells, and the expression was slow with the vector GV341. Virus, then lentivirus infection to establish 19 mixed clone stable cell lines, including empty vector control (NC), HER2WT, HER2YVMAdup, P780_Y781insGSP, G776>VC, V777L, L755S, D769H, G776R, G776C, L755P, V842I, L866M, R896C, S310F, S310Y, G309A, G309E and D769Y. All cell cultures were supplemented with DMEM/F12 medium plus 5% horse serum, 20 ng/ml EGF, 10 μg/ml insulin, 0.5 μg/ml hydrocortisone, 1% penicillin/streptomycin (P/S) and 100 ng/ml Cholera Toxin.

3, reagents and instruments

DMEM/F12 (Gibco, 10-092-CVR), horse serum (source leaf organism, MP20006), insulin (source leaf organism, 11070-73-8), epidermal growth factor (Peprotech, AF-100-15-100) Cholera toxin (sigma, 9012-63-9),

Hydrocortisone (source leaf organism, 50-23-7), trypsin (Gibco, 25200-072), Puromycin (翊圣, 60210ES25), RIPA lysate (Dingguo, WB-0071), MTT (Genview, JT343).

4. Test method

The inhibitory effect of drugs on the proliferation and growth of tumor cells was detected by MTT susceptibility test. 10 concentration points for each drug, 1 detection time point, 2 double holes. Inoculate 2000 cells per well in a 96-well culture plate (100 μl/well of medium), add the appropriate concentration of the drug (0.00128-500 nM for compound A; 0.00128-500 μM for lapatinib), and set a duplicate hole for each concentration. Set the appropriate concentration of the vehicle control. Tumor cells were cultured for 72 h at 37 ° C under 5% CO 2 . Then use MTT test. The absorbance of each well was measured by an enzyme-linked immunosorbent assay at OD490nm, and the cell growth inhibition rate was calculated by the following formula:

Inhibition rate = (OD value control hole-OD value administration hole) / OD value control hole × 100%

The half-inhibitory concentration IC50 was calculated according to the nonlinear regression method according to the respective concentration inhibition rates.

5, test results

The results showed that Compound A had a good inhibitory effect on HER2 mutant lung adenocarcinoma cells, while the control compound lapatinib had a poor inhibitory effect. The specific results are shown in Table 3:

Table 3 Compound A mutant HER2 in vitro cell proliferation inhibition IC ₅₀ are summarized

Example 4: A compound A treatment of advanced non-small cell lung adenocarcinoma with HER2 mutation

Test method: The enrolled subjects were pathologically diagnosed patients with advanced non-small cell lung adenocarcinoma, and confirmed the presence of HER2 gene mutation (using the human HER2 gene detection kit fluorescence of Xiamen Aide Biomedical Technology Co., Ltd.) The PCR method (1st to 7th, 10th) and the NGS second-generation sequencing method (8th, 9th, and 11th cases) were performed on the tumor pathological section of the patient. Subjects who met the enrollment requirements were given Compound A at 320 mg/d and/or 400 mg/d orally once daily for continuous administration until the patient developed disease progression/intolerance. As of August 2016, 11 patients with advanced non-small cell lung adenocarcinoma with HER2 mutations had been enrolled. The average age of the subjects was 58.4 years. All subjects continued to progress after treatment with different means, such as docetaxel or pemetrexed chemotherapy, or targeted drugs such as gefitinib and afatinib. Adverse events were grade 1 to 2, including 1/2 grade diarrhea (4 cases), grade 2 fatigue (2 cases), grade 1 rash (2 cases) and grade 1 dyspnea (1 case), which were clinically controllable. There were no serious adverse events (SAE) and discontinuation due to adverse events, dose reduction, and early out-of-group. Six patients (54.5%) received PR, 3 patients (27.3%) had stable disease, and 2 patients (18.2%) had PD, ORR was 54.5%, and DCR was 81.8%. The median PFS was 6.2 months (95% CI 1.23-11.57). There are still 5 patients in the group treatment.

The above test results suggest that in patients with HER2 mutation in non-small cell lung adenocarcinoma, the compounds A320mg/d and 400mg/d are safe, well tolerated, clinically controllable, and have significant antitumor effects.

Details of patients who have been evaluated for efficacy are shown in Table 4 below. Among them, the meaning of each English abbreviation is as follows.

PD: disease progression, increase in diameter and minimum of the target lesion diameter by at least 20% and absolute increase in diameter sum by at least 5 mm (one or more new lesions are also considered disease progression); SD: disease stabilization, target The sum of the maximum diameters of lesions did not reach PR, or increased PD PR: partial remission, and the sum of target lesion diameters was reduced by at least 30% from baseline, at least for 4 weeks. UK: Unknown.

Claims (14)

1. Use of a compound of formula A or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a HER2 mutant cancer,

   
2. The use according to claim 1, wherein the cancer is selected from the group consisting of lung cancer, breast cancer and gastrointestinal cancer.
3. The use according to claim 2, wherein the lung cancer is non-small cell lung cancer.
4. The use according to claim 2, wherein the lung cancer is a lung adenocarcinoma.
5. The use according to claim 2, wherein the lung cancer is advanced lung cancer.
6. The use according to claim 1, wherein the cancer is a cancer that progresses after chemotherapy, radiation therapy or targeted therapy.
7. The use according to claim 6, wherein said chemotherapy is selected from the group consisting of an alkylating agent, a platinum complexing agent, a metabolic antagonist, a plant alkaloid, a hormone anticancer agent, a proteasome inhibitor, and an aromatase inhibitor. One or more of them; preferably one or more selected from the group consisting of carboplatin, cisplatin, oxaliplatin, 5-fluorouracil, vinorelbine, gemcitabine, pemetrexed or docetaxel Perform chemotherapy.
8. The use according to claim 6, wherein the targeted therapy is treatment with one or more selected from the group consisting of an EGFR inhibitor and a VEGFR inhibitor.
9. The use according to claim 8, wherein the EGFR inhibitor is selected from one or more of gefitinib, erlotinib, ectinib and afatinib.
10. The use according to claim 8, wherein the VEGFR inhibitor is selected from one or more of sunitinib, apatinib and faritinib.
11. The use according to claim 1, wherein the pharmaceutically acceptable salt of Compound A is a maleate salt, preferably a dimaleate salt.
12. The use according to claim 1, wherein the compound A or a pharmaceutically acceptable salt thereof is used in a compound amount of from 1 mg/kg to 20 mg/kg, preferably from 2 mg/kg to 10 mg/kg, more preferably 4, based on the compound A. ~8mg/kg.
13. The use according to claim 1, wherein the compound A or a pharmaceutically acceptable salt thereof is used in a compound amount of from 100 mg to 1000 mg, preferably from 240 to 560 mg, more preferably from 320 to 480 mg, based on the compound A.
14. The use according to any one of claims 1 to 13, wherein the compound A or a pharmaceutically acceptable salt thereof is prepared into a composition, which further contains a pharmaceutically acceptable carrier.