WO2022186649A1 - Anticancer drug use of phthalazinone derivative as cancer treatment agent for patients with resistance to parp inhibitor - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing a phthalazinone derivative as an active ingredient for treating PARP inhibitor-resistant cancer. More specifically, the present invention relates to a pharmaceutical composition for preventing or treating PARP inhibitor-resistant cancer, the composition containing a phthalazinone derivative of chemical formula 1 as an active ingredient. The pharmaceutical composition containing the phthalazinone derivative according to the present invention exhibits an effective antiapoptotic effect on PARP inhibitor-resistant cancer cells.

Description

Anticancer drug use of phthalazinone derivatives as a cancer treatment for patients with resistance to PARP inhibitors

The present invention relates to a pharmaceutical composition containing a phthalazinone derivative as an active ingredient for the treatment of human poly(ADP ribose) polymerase inhibitor (PARP inhibitor)-resistant cancer. More specifically, the present invention relates to a compound of Formula 1 below 4-[3-(3-[(cyclopropylamino)methyl]azetidine-1-carbonyl)-4-fluorobenzyl]phthalazine-1(2H )-On or a pharmaceutically acceptable salt thereof, as an active ingredient, relates to a pharmaceutical composition for preventing or treating PARP inhibitor-resistant cancer, and to a method for preventing or treating PARP inhibitor-resistant cancer using the composition .

Some cancer patients achieve a complete response (complete remission) to conventional therapies such as radiation therapy and/or chemotherapy and targeted therapy, but the majority of cancer patients are moderate and non-response; The disease progresses in most cancer patients. Progression of the disease also occurs in a certain number of cancer patients who have received such treatment. Attempts to treat some advanced cancer patients with these conventional therapies or with maintenance therapy are limited.

The relatively rapid acquisition of resistance to cancer therapeutics remains a major obstacle to successful cancer treatment. Although mutations conferring some specific resistance have indeed been shown to represent acquired drug resistance in many cancer patients, the relative contribution of mutations and non-mutant mechanisms to drug resistance and the role of tumor cell subpopulations are somewhat unclear.

New therapeutic approaches are needed to successfully address heterogeneity within cancer cell populations and the emergence of treatment-resistant cancer cells.

Olaparib is human poly (ADP ribose) polymerase [Poly (ADP Ribose) Polymerase; PARP-1, PARP-2 and PARP-3; Hereinafter, collectively referred to as 'PARP')], it inhibits the proliferation of specific tumor cell lines in vitro and tumor growth in vivo as a monotherapy or combination therapy with established chemotherapy.

PARP is a family of enzymes involved in DNA repair and consists of 18 components. These enzymes are located in the cell nucleus and are involved in the repair of DNA damage. PARP inhibitors have been used to target BRCA1/2-defective cells that undergo DNA single-strand breaks (SSBs).

The BRCA1 or BRCA2 gene is responsible for homologous-mediated repair of damaged double-stranded DNA. However, the use of PARP in cells in which the BRCA1/2 gene is deleted increases these DNA single-strand breaks (SSBs) in the presence of a PARP inhibitor, and these single-strand breaks (SSBs) cause irreversible toxic double-strand breaks during replication ( DSBs: double-strand breaks, eventually leading to cell death due to genomic instability caused by an impaired homologous-mediated repair system.

The DNA-binding domains of both PARP-1 and PARP-2 contain two zinc finger motifs that promote migration of repair enzymes to the site of DNA damage. PARP-1 and PARP-2 catalyze the transfer of ADP-ribose units from nicotinamide adenine dinucleotide (NAD+) to nuclear receptor proteins, resulting in the formation of ADP-ribose polymers. This is a key process for repairing DNA damage caused by DNA-damaging chemotherapeutic agents and radiation through base-excision repair (BER)-mediated single-strand break repair. Therefore, PARP inhibition causes the death of cancer cells with defective DNA damage repair mechanisms.

As an example of a PARP inhibitor, olaparib (Olaparib, AZD-2281), rucaparib (Rucaparib, AG-014699), niraparib (Niraparib, MK-4827), talazoparib (Talazoparib, BMN-673), veliparib ( Veliparib, ATB-888), Fluzoparib (SHR-3162), Pamiparib (BGN-290), Senaparib (Senaparib, IMP-4297), Stenoparib (Stenoparib, 2X-121), AZD -5305 etc. are mentioned.

On the other hand, US Patent No. 9,682,973 discloses that the compound of Formula 1 below has anticancer activity as a PARP inhibitor 4-[3-(3-[(cyclopropylamino)methyl]azetidine-1-carbonyl)-4-fluorobenzyl]p Thalazin-1(2H)-one, or a pharmaceutically acceptable salt thereof, is disclosed. In particular, the hydrochloride salt of the compound of Formula 1 is one of the promising therapeutic drug candidates as anticancer agents.

PARP inhibitors known as such conventional effective anticancer agents are also known to cause resistance to cancer treatment, thereby becoming an obstacle to successful cancer treatment. It is known that such PARP inhibitor-induced resistance is caused by various mechanisms (Cancers 2020, 12, 1607; doi:10.3390/cancers12061607). Among these various resistance development mechanisms, a reversion mutation of the BRCA2 gene is known to be the main cause of PARP inhibitor resistance (Pettitt et al. CANCER DISCOVERY , October, 2020; DOI: 10.1158/2159-8290.CD-19). -1485 ). It is also disclosed that backmutations in these genes, such as BRCA, are the mechanism of acquired resistance observed against several clinically tested PARP inhibitors, such as olaparib, thalazoparib, and rucaparib (Lin et al. Cancer Discov , February , 2019; DOI: 10.1158/2159-8290.CD-18-071 ).

[Prior art literature]

[Patent Literature]

US Patent 9,682,973

[Non-patent literature]

Cancers 2020, 12, 334; doi: 10.3390/cancers12020334

Cancers 2020, 12, 1607; doi:10.3390/cancers12061607

Petittt et al. CANCER DISCOVERY , October, 2020; DOI: 10.1158/2159-8290.CD-19-1485

Lin et al. Cancer Discov, February, 2019; DOI: 10.1158/2159-8290.CD-18-071

The present inventors have identified the compound of formula 1 4-[3-(3-[(cyclopropylamino)methyl]azetidine-1-carbonyl)-4-fluorobenzyl]phthalazin-1(2H)-one Or, the present invention was completed by confirming that a pharmaceutically acceptable salt thereof can be usefully used not only for various cancers but also for the treatment of cancers resistant to PARP inhibitors, including olaparib.

An object of the present invention is the compound of formula 1 4-[3-(3-[(cyclopropylamino)methyl]azetidine-1-carbonyl)-4-fluorobenzyl]phthalazin-1(2H)-one Or to provide a pharmaceutical composition for preventing or treating cancer with resistance to a PARP inhibitor, containing a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a method for preventing or treating cancer having resistance to a PARP inhibitor, comprising administering the composition to a subject.

In one aspect for achieving the above object, the present invention provides a compound of Formula 1 4-[3-(3-[(cyclopropylamino)methyl]azetidine-1-carbonyl)-4-fluorobenzyl ]Provides a pharmaceutical composition for preventing or treating PARP inhibitor-resistant cancer, containing phthalazin-1(2H)-one as an active ingredient.

Also, a salt form of the compound of Formula 1 (Compound 1) may be included in the scope of the present invention. For example, inorganic acids (hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, etc.), organic carboxylic acids (acetic acid, haloacetic acid such as trifluoroacetic acid, propionic acid, maleic acid, succinic acid, malic acid, citric acid, tartaric acid, salicylic acid, etc.) ), and acidic sugars (glucuronic acid, galacturonic acid, gluconic acid, ascorbic acid, etc.), acidic polysaccharides (hyaluronic acid, chondroitin sulfate, arginic acid, etc.), or sulfonic acids, such as chondroitin sulfate, sugar esters organic sulfonic acids (methanesulfonic acid, p-toluenesulfonic acid, etc.) may be included.

In the present invention, the term "solid cancer" has characteristics distinguishing it from blood cancer, and includes bladder, breast, intestine, kidney, lung, liver, brain, esophagus, gallbladder, ovary, pancreas, stomach, cervix, and thyroid gland. It is a cancer consisting of a mass caused by abnormal cell growth in various solid organs such as , prostate and skin.

In the present invention, the term "PARP inhibitor-resistant cancer" refers to a cancer in which resistance is induced by repeated and long-term use of a PARP inhibitor, an anticancer agent that has been mainly used for cancer treatment such as solid cancers such as ovarian cancer and breast cancer, and blood cancer. When a PARP inhibitor, which initially exhibits a clear therapeutic effect by inhibiting cell division of cancer cells, is continuously administered to cancer patients for a long period of time, the therapeutic effect decreases. because it does Cancers resistant to PARP inhibitors in the present invention include, but are not limited to, breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, stomach cancer, blood cancer, and the like.

In the present invention, the term “PARP inhibitor” is an abbreviation for human poly(ADP ribose) polymerase inhibitor, and includes PARP inhibitors known in the art and PARP inhibitors to be developed later. PARP inhibitors include Olaparib (AZD-2281), Rucaparib (AG-014699), Niraparib (MK-4827), Talazoparib (BMN-673), Veliparib (Veliparib, ATB) -888), fluzoparib (SHR-3162), pamiparib (Pamiparib, BGN-290), senaparib (Senaparib, IMP-4297), stenoparib (Stenoparib, 2X-121), AZD-5305, etc. However, it is not limited thereto.

In the present invention, the term "prevention" refers to any action that inhibits or delays the occurrence, spread, and recurrence of cancer by administration of the pharmaceutical composition according to the present invention, and "treatment" refers to any activity that causes suspicion of cancer by administration of the pharmaceutical composition. And it means any action that improves or beneficially changes the symptoms of the affected individual.

Cancers that can be prevented or treated by the composition of the present invention include breast cancer, ovarian cancer, pancreatic cancer, prostate cancer, stomach cancer, blood cancer, and the like, preferably olaparib-resistant cancer, but is not limited thereto.

According to a specific embodiment of the present invention, the composition of the present invention, for example, the hydrochloride of the compound of Formula 1 shows resistance to olaparib cancer cell lines (SNU 601/Olaparib-resistant cells, SNU-638/Olaparib-resistant cells, SNU- 668/Olaparib-resistant cells, MCF7/Olaparib-resistant cells, MDA-MD-468/Olaparib-resistant cells, SNU-5126) were confirmed to exhibit a concentration-dependent antiproliferative effect (FIG. 3) . In addition, it was confirmed that the hydrochloride of the compound of Formula 1 exhibits an antiproliferative effect in a concentration-dependent manner against the obtained cancer cell line (HCC1428), which has a conventional BRCA2 back mutation and is known to have resistance to a PARP inhibitor (Fig. 2 and Table 5). Considering that the conventional BRCA2 reversion mutation is known to be the main mechanism for inducing resistance of cancer cells to PARP inhibitors, the hydrochloride salt of the compound of Formula 1 can be widely applied not only to olaparib-resistant cancers, but also to PARP inhibitor-resistant cancers. It is expected.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier. As used herein, the term “pharmaceutically acceptable” means exhibiting non-toxic properties to cells or humans exposed to the composition. The carrier may be used without limitation as long as it is known in the art, such as buffers, preservatives, softening agents, solubilizers, isotonic agents, stabilizers, bases, excipients, lubricants, and the like.

In addition, the pharmaceutical composition of the present invention can be used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. have. Furthermore, it may be used in the form of an ointment, lotion, spray, patch, cream, powder, suspension, gel, or gel-form external preparation for skin. Carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient, for example, starch, calcium carbonate (calcium) in the phthalazinone derivative of Formula 1 above. carbonate), sucrose or lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. may be used.

On the other hand, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not to cause side effects, and the effective dose level depends on the patient's health condition, Cancer type, severity, drug activity, sensitivity to drug, administration method, administration time, administration route and excretion rate, treatment period, factors including drugs used in combination or concomitantly, and other factors well known in the medical field. can The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the phthalazinone derivative compound of Formula 1 contained in the composition of the present invention may vary depending on the age, sex, and weight of the patient. In general, the compound of Formula 1 (Compound 1) of the present invention contains a solid content 0.1 to 1000 mg per day, preferably 1 to 500 mg, more preferably 1 to 300 mg daily or every other day, intermittently, or divided into 1 to 3 times a day may be administered. However, the dosage may be increased or decreased depending on the route of administration, disease severity, sex, weight, age, etc., so the dosage does not limit the scope of the present invention in any way.

In another aspect, the present invention provides a method for preventing or treating PARP inhibitor-resistant cancer, comprising administering a compound of Formula 1 or a pharmaceutically acceptable salt thereof to an individual in need thereof.

As used herein, the term "individual" refers to monkeys, cows, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits or guinea pigs, including humans, who have or can develop the cancer. It means all animals, including, and can effectively prevent or treat the cancer by administering the pharmaceutical composition of the present invention to an individual. The pharmaceutical composition of the present invention may be administered in parallel with a conventional therapeutic agent.

The term "administration" of the present invention means providing a predetermined substance to a patient by any suitable method, and the administration route of the composition of the present invention may be administered through any general route as long as it can reach the target tissue. have. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, may be administered intrarectally, but is not limited thereto. In addition, the pharmaceutical composition of the present invention may be administered by any device capable of transporting an active substance to a target cell. Preferred administration modes and formulations are intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, drip injections, and the like. For injection, aqueous solvents such as physiological saline and Ringer's solution, vegetable oil, higher fatty acid esters (eg, ethyl oleate), and non-aqueous solvents such as alcohols (eg, ethanol, benzyl alcohol, propylene glycol, glycerin, etc.) Stabilizers for preventing deterioration (e.g., ascorbic acid, sodium hydrogen sulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH control, to inhibit microbial growth Pharmaceutical carriers such as preservatives (eg, phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.) may be included.

The pharmaceutical composition of the present invention may further include a known anticancer agent in addition to the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and may be used in combination with other known treatments for the treatment of these diseases. Other treatments include, but are not limited to, chemotherapy, radiation therapy, hormone therapy, bone marrow transplantation, stem-cell replacement therapy, other biological therapies, immunotherapy, and the like.

Examples of anticancer agents that may be included in the pharmaceutical composition of the present invention include mechloethamine, chlorambucil, phenylalanine, mustard, cyclophospha as DNA alkylating agents. Cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), streptozotocin, busulfan, thiotepa, cisplatin ( cisplatin) and carboplatin; oxaliplatin, fluorouracil (5-Fluorouracil, 5-FU), gemcitabine, dactinomycin (actinomycin D) as anti-cancer antibiotics, doxorubicin (adriamycin), liposomal doxorubicin, daunorubicin, idarubicin, mitoxantrone, plicamycin, mitomycin C and bleomycin; And as plant alkaloids vincristine, vinblastine, vinorelbine, eribulin, paclitaxel, docetaxel, lead-paclitaxel (nab-paclitaxel) ), etoposide, teniposide, topotecan and irinotecan, and the like, but are not limited thereto.

In another embodiment, the present invention provides compound 4-[3-(3-[(cyclopropylamino)methyl]azetidine-1-carbonyl) of Formula 1 for use in the prevention or treatment of PARP inhibitor-resistant cancer -4-fluorobenzyl]phthalazin-1(2H)-one, or a pharmaceutical use thereof.

In the details of this aspect, the above description of the pharmaceutical composition according to the one aspect and the above description of the method for preventing or treating PARP inhibitor-resistant cancer according to another aspect may be applied as it is.

According to the present invention, the composition containing the phthalazinone derivative of Formula 1 as an active ingredient exhibits an effective killing effect of PARP inhibitor-resistant cancer cells. Therefore, the composition containing the compound of Formula 1 (Compound 1) or a pharmaceutically acceptable salt thereof of the present invention as an active ingredient can be usefully used as a pharmaceutical composition for the prevention or treatment of PARP inhibitor-resistant cancer.

1 is a graph showing the cell viability (%) and antiproliferative effect according to the concentration of the hydrochloride of Compound 1 in breast cancer cell lines and gastric cancer cell lines.

1a is a graph showing the cell viability (%) results according to the concentration of the hydrochloride of Compound 1 in breast cancer cell lines and gastric cancer cell lines.

Figure 1b is a diagram showing the anti-proliferative effect according to the concentration of the hydrochloride of Compound 1 in breast cancer cell lines and gastric cancer cell lines.

Figure 2a is a graph showing the cell viability (%) results according to the concentration of the hydrochloride of Compound 1 in olaparib-resistant cell lines.

Figure 2b is a graph showing the cell viability (%) results according to the concentration of the hydrochloride of Compound 1 in a breast cancer cell line having a BRCA2 back mutation.

Figure 3a is a diagram showing the anti-proliferative effect according to the concentration of the hydrochloride of Compound 1 in olaparib-resistant cell lines.

Figure 3b is a diagram showing the anti-proliferative effect in the cell line derived from the PDX model of olaparib-resistant patients according to the hydrochloride concentration of Compound 1.

Figure 3c is a diagram showing the antiproliferative effect according to the concentration of the hydrochloride of Compound 1 in a breast cancer cell line having a BRCA2 back mutation.

In the present invention, the term "compound of formula 1" is also simply referred to as "compound of formula 1", "compound 1" or "compound of the present invention". Similarly, “salt of compound of formula 1” is also referred to as “salt of compound of formula 1”, “salt of compound 1” or “salt compound of the present invention”. For example, when the compound of Formula 1 exists in the form of a hydrochloride salt, it may be referred to as "hydrochloride salt of the compound of formula 1", "hydrochloride salt of compound 1", or "hydrochloride compound of the present invention".

As used herein, the term “pharmaceutically acceptable salt” refers to the free base form of the compound of Formula 1 can be prepared by any suitable method in the art. For example, the free base can be combined with an inorganic acid (eg, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, phosphoric acid, etc.), an organic acid (eg acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid). , malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyl acid (such as glucuronic or galacturonic acid), alpha hydroxyl acid (such as citric or tartaric acid), amino acids (such as aspartic or glutamic acid), It can be prepared by treatment with an aromatic acid (such as benzoic acid or cinnamic acid), a sulfonic acid (such as p-toluenesulfonic acid or ethanesulfonic acid), etc. The hydrochloride salt of the compound of formula (1) is most preferred.

The compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof can be easily obtained by, for example, the method described in US Patent No. 9,682,973, or by preparation according thereto.

The compound of Formula 1 or a pharmaceutically acceptable salt thereof exhibits PARP inhibitory activity and can be usefully used for cancer treatment.

In the pharmaceutical composition of the present invention, one or more pharmaceutically acceptable excipients are suitably included in the range capable of achieving the effects of the present invention.

As used herein, the term "excipient" refers to a pharmaceutically acceptable inert ingredient used in the manufacture of pharmaceutical formulations. Excipients include, but are not limited to, for example, diluents, binders, disintegrants, super-strong disintegrants, glidants, pH adjusters, lubricants, fillers, stabilizers, antioxidants, film coatings. The excipient used in the present invention may be appropriately selected by referring to literature known in the art, for example, Handbook of Pharmaceutical Excipients, Sixth edition, Pharmaceutical Press, which is incorporated herein by reference.

Hereinafter, the present invention will be described in more detail through examples. However, the following examples are for illustrating the present invention, and the present invention is not limited by the following examples.

Materials and Methods

[Cell Culture]

The TNBC breast cancer cell line, MDA-MB-468, was purchased from ATCC and cultured according to the supplier's instructions. MCF-7 and HCC1428 were luminal breast cancer cell lines, and SNU-601, SNU-638, and SNU-668 were gastric cancer cell lines, purchased from KCLB and cultured according to the supplier's instructions. SNU-5126 cells were obtained from a patient with BRCA2 580del4 mutation and HR-positive, HER2-negative IDC breast cancer and resistant to olaparib (Accession No.: KCLRF-BP-00510). Briefly, each cell was cultured in RPMI1640 medium supplemented with 10% fetal bovine serum (FBS) and 10 μg/ml gentamicin at 37° C. and 5% CO ₂ conditions. Information on the cell lines used in Examples is shown in Table 1.

Example 1: Evaluation of the anticancer effect of the hydrochloride of Compound 1

Example 1-1: Confirmation of doubling time of cell lines

The cell line was aliquoted in a 6-well plate with a cell number of 1 x 10 ⁵ and then cultured overnight at 37° C. and 5% CO ₂ conditions. Cell lines were measured every 24 or 48 hours. The daily growth rate was normalized to the value of the first day. The doubling times of the cell lines are shown in Table 2.

Cell culture was carried out for 11 days (corresponding to 5 or more cell divisions), the average doubling time of cells was 47 hours, and it was confirmed that the cell line was in a state suitable for the experiment for 11 days.

Example 1-2: Evaluation of the anti-proliferative effect of the hydrochloride salt of Compound 1: CFA analysis

A designated number of cells was aliquoted into a 6-well plate and incubated at 37° C. in 5% CO ₂ for 36 hours. After incubation, the cells were treated with various concentrations of the hydrochloride salt of the compound of formula 1 (the hydrochloride of compound 1) for 11 days (a time period of 5 or more cell divisions). After removing the culture medium, the cells were washed with phosphate buffered saline (PBS) and stained with 0.1% Coomassie blue solution (Sigma Aldrich) for 1 hour at room temperature. In the next step, excess staining solution was removed and the plates were washed with PBS and air dried. Cell colonies were colony counter (Bid-Rad, California, USA); Cell viability and IC ₅₀ of hydrochloride of compound 1 were determined using SigmaPlot software version 10.0 (SPSS). Each experiment was repeated three times.

The cell viability (%) results according to the concentration of the hydrochloride of Compound 1 in breast cancer cell lines and gastric cancer cell lines are shown in Table 3 and FIG. 1 .

Example 2: Evaluation of the anticancer effect of compound 1 hydrochloride in olaparib-resistant breast cancer and gastric cancer, and breast cancer with BRCA2 back mutation

Olaparib, a PARP inhibitor, is a drug used for the treatment of ovarian cancer and breast cancer. However, long-term administration of PARP inhibitors such as olaparib induces resistance in cancer cells that do not respond to PARP inhibitors.

Example 2-1: Establishment of olaparib-resistant breast cancer and gastric cancer cell lines

Breast cancer and gastric cancer cell lines were exposed to olaparib concentrations from 0.1 μM to 10 μM for 7 months while gradually increasing concentrations to establish olaparib-resistant cell lines. When the olaparib-resistant cell line is cultured in an incubator at 37°C and 5% carbon dioxide concentration by adding 10% fetal bovine serum and 10ug/mL gentamicin to RPMI1640 medium, 10 μM olaparib is produced by colony forming (CFA). assay) was defined as when the cell viability was reduced to less than 20% for 14 days by the method, and the establishment of a resistant cell line was verified through DNA fingerprinting analysis.

Example 2-2: Confirmation of anti-proliferative effect of olaparib-resistant breast cancer and gastric cancer, and breast cancer with BRCA2 reversion mutation

[Correction by Rule 91 20.04.2022]
0, 0.001, 0.01, 0.05, 0.1, 0.25, 0.5, 1, 2.5, 5, 10 μM of the hydrochloride salt of the compound of Formula 1 in each of the olaparib-resistant breast cancer and gastric cancer cell lines pretreated with olaparib in the method of Example 2-1 After treatment with a concentration of It was confirmed through the CFA performance result that it exhibits an excellent anti-proliferative effect (FIG. 3). The IC ₅₀ of the hydrochloride salt of the compound of Formula 1 against olaparib-resistant cell lines is shown in Table 5 and FIG. 2a below. In addition, the same experiment as in Example 1-2 was also performed on the cancer cell line HCC1428 (source: Korea Cell Line Research Foundation KCLB) known to have conventional BRCA2 reversion mutation and PARP inhibitor resistance, and the results are shown in Table 5 and FIG. 2b below. , are shown together in 3c.

Hydrochloride of Compound 1 effectively inhibited cell proliferation activity at an acceptable concentration (average steady-state blood concentration confirmed in clinical trials of hydrochloride of Compound 1: 1.4 to 5.6 μM) in cell lines that acquired resistance to olaparib. Steady state refers to a state in which the plasma concentration is maintained at a constant value because the dose and removal amount of the drug are the same.

As a result, compared with non-resistant breast cancer (MCF7/parental cells, MDA-MB-468/parental cells) and gastric cancer (SNU-601/parental cells, SNU-638/parental cells, SNU-668/parental cells), , Olaparib-resistant breast cancer cell line (MCF7/Olaparib-resistant cells, MDA-MB-468/Olaparib-resistant cells, SNU-5126) and olaparib-resistant gastric cancer cell line (SNU-601/Olaparib-resistant cells, SNU-638/Olaparib) -resistant cells, SNU-668/Olaparib-resistant cells), it was confirmed that the hydrochloride of compound 1 still showed anticancer efficacy. In addition, it was confirmed that the hydrochloride of Compound 1 showed anticancer efficacy against HCC1428, a cancer cell line known to have resistance to conventional BRCA2 reversion mutations and PARP inhibitors.

[Correction by Rule 91 20.04.2022]

Claims (5)

1. A pharmaceutical for the prophylaxis or treatment of human poly(ADP ribose) polymerase inhibitor (PARP inhibitor)-resistant cancer, comprising as an active ingredient a compound of Formula 1 or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients Composition:

   
2. According to claim 1, wherein the PARP inhibitor is olaparib (Olaparib, AZD-2281), rucaparib (Rucaparib, AG-014699), niraparib (Niraparib, MK-4827), talazoparib (Talazoparib, BMN-673) , Veliparib (ATB-888), Fluzoparib (Fluzoparib, SHR-3162), Pamiparib (BGN-290), Senaparib (Senaparib, IMP-4297), Stenoparib (Stenoparib, 2X- 121), and any combination thereof.
3. The pharmaceutical composition of claim 1, wherein the PARP inhibitor is olaparib.
4. The pharmaceutical composition according to claim 1, wherein the active ingredient is the hydrochloride salt of the compound of formula (1).
5. A method for preventing or treating PARP inhibitor-resistant cancer, comprising administering the pharmaceutical composition of any one of claims 1 to 4 to a subject other than a human.

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