Therapeutic agent for PARP inhibitor-resistant cancer

Abstract

To provide a therapeutic agent for PARP inhibitor-resistant cancer.SOLUTION: The present invention relates to a pharmaceutical composition for treating or preventing a solid cancer patient having resistance to a PARP inhibitor, and the pharmaceutical composition according to the present invention can effectively reduce the size of a tumor in a patient having resistance to a PARP inhibitor.SELECTED DRAWING: Figure 4

Description

[Technical Field]

[0001] This application claims the benefit of priority based on Korean Patent Application No. 10-2021-0064278 dated May 18, 2021, and all contents disclosed in the documents of said Korean patent application are incorporated herein by reference.

[0002] The present invention relates to a cancer therapeutic agent that can be used to treat patients with solid cancers that are resistant to PARP inhibitors. [Background technology]

[0003] The accumulation of DNA mutations is known to be one of the major causes of cancer development. Mammals grow and develop through continuous cell division from a single cell, the fertilized egg. During this process, DNA mutations (hereinafter referred to as "DNA damage") inevitably occur. DNA damage is repaired using various DNA repair mechanisms, such as homologous recombination (HR) and non-homologous end joining (NHEJ). Each DNA repair mechanism involves various proteins, and if mutations occur in some of these proteins, problems arise in the DNA repair mechanism, increasing the risk of cancer development by several to several hundred times. Generally, loss of homologous recombination function leads to genomic instability, which causes various genetic changes and ultimately leads to tumor development.

[0004] The BRCA1 / 2 genes, involved in repairing damaged DNA, suppress tumorigenesis. Mutations in the BRCA1 / 2 genes impair their function, preventing proper repair of damaged DNA and resulting in cumulative DNA damage, leading to cancer. This condition is known as homologous recombination deficiency (HRD). Breast cancer and ovarian cancer, both of which are well-known as homologous recombination-deficient tumors, are associated with BRCA1 / 2 gene mutations. Women with BRCA1 / 2 gene mutations are known to have an increased risk of developing breast cancer and ovarian cancer, up to 80% and 60%, respectively. However, BRCA1 / 2 gene mutations are also associated with gastric cancer, pancreatic cancer, prostate cancer, gallbladder cancer, biliary tract cancer, and colorectal cancer, in addition to breast cancer and ovarian cancer.

[0005] Poly(ADP-ribose) polymerase (PARP) proteins are essential for repairing errors that inevitably occur during DNA replication. They are activated by recognizing damaged DNA in the nucleus and then activate DNA repair-related proteins through a post-translational process (PARylation). To date, 17 members of the PARP family have been identified, but only PARP1 / 2 have been identified as DNA repair enzymes with poly(ADP-ribosylation) activity, and are known to be essential for cell survival.

[0006] Homologous recombination-deficient tumors are known to be sensitive to DNA damage induced by PARP inhibitors. Therefore, PARP inhibitors have great potential as cancer therapeutic agents in clinical settings. Indeed, Olaparib (Lynparza) TM ), rucaparib (Rucaparib: Rubraca TM ), Niraparib (ZEJULA TM ), Talazoparib (Talzenna TMPARP inhibitors such as niraparib are prescribed to patients with ovarian cancer, breast cancer, or prostate cancer who have BRCA1 / 2 mutations (germ-line mutations), and in particular, niraparib is used as a maintenance therapy for recurrent epithelial ovarian cancer, advanced serous ovarian cancer (fallopian tube cancer, or primary peritoneal cancer), etc. that have completely or partially responded to platinum-based anticancer chemotherapy.

[0007] 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile, which has been developed as a PARP inhibitor, has the structure of the following chemical formula 1. The compound of the following chemical formula 1 or a pharmaceutically acceptable salt thereof exhibits inhibitory activity against not only PARP1 / 2 but also tankyrase1 / 2. Tankyrase is known to be involved in the Wnt / β-catenin signaling pathway, DNA repair processes, and mitosis, which is closely related to the cell cycle. In addition, tankyrase1 / 2 ADP-ribosylates TRF-1, functioning as a positive regulator of telomere length and promoting telomere elongation by telomerase. In addition, the compound of the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof is expected to be effective in treating recurrent epithelial ovarian cancer, high-grade serous ovarian cancer, and the like that respond completely or partially to platinum-based anticancer chemotherapy.

[0008] <Chemical formula 1> TIFF2026026362000002.tif2664

[0009] While various PARP inhibitors have potential and promise as targeted therapeutic agents for cancer treatment, PARP inhibitors, including olaparib, have a high rate of innate / acquired resistance or refractory disease, just like other anticancer drugs. Drug resistance is becoming more prevalent in homologous recombination-deficient tumors, and research into this issue is ongoing, but no significant progress has been made to date.

[0010] Furthermore, it is not known whether the compound of Chemical Formula 1 can treat solid cancers resistant to anticancer drugs other than Chemical Formula 1, particularly solid cancers resistant to PARP inhibitors, which are used in conventional standard treatments.

[0011] Under such circumstances, the inventors of the present invention have extensively studied anticancer agents that can be used to treat patients who are resistant to PARP inhibitors, and as a result, have confirmed that a compound comprising Chemical Formula 1 of the present invention reduces tumor size in patients who are resistant to conventional PARP inhibitors (such as olaparib), thereby completing the present invention. [Prior art documents] [Patent documents]

[0012] [Patent Document 1] Korean Patent Registration No. 10-1136702 (Publication Date: 2012.4.20) [Patent Document 2] Korean Patent Registration No. 10-1146806 (Publication Date: 2012. 5. 22.) [Patent Document 3] Korean Patent Registration No. 10-1837047 (Publication Date: 2018.3.09.) Summary of the Invention [Problem to be solved by the invention]

[0013] An object of the present invention is to provide a pharmaceutical composition comprising 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof as a composition for treating solid tumors in patients resistant to PARP inhibitors.

[0014] Another object of the present invention is to provide a method for treating solid tumors in an individual having resistance to PARP inhibitors by administering 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof to the individual.

[0015] The present invention aims to provide 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof for use in treating solid tumors in patients with resistance to PARP inhibitors. [Means for solving the problem]

[0016] To achieve the above object, the present invention provides a pharmaceutical composition for treating solid tumors in patients with resistance to PARP inhibitors, comprising 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof.

[0017] In one embodiment of the present invention, the PARP inhibitor may be one or more selected from, but not limited to, olaparib, rucaparib, niraparib, and talazoparib.

[0018] In one embodiment of the invention, the patient may have a BRCA1 / 2 mutation. In one embodiment of the present invention, the patient may have a germline BRCA1 / 2 mutation.

[0019] In one embodiment of the present invention, the patient may have a somatic BRCA1 / 2 mutation. In one embodiment of the present invention, the patient may have a BRCA1 / 2 mutation and initially responded to a PARP inhibitor, but developed resistance during treatment and did not respond to the PARP inhibitor, or the cancer may have recurred.

[0020] In one embodiment of the present invention, the patient may have a BRCA1 / 2 mutation but has not responded to a PARP inhibitor.

[0021] In one embodiment of the present invention, the patient may not have a BRCA1 / 2 mutation and may have previously failed to respond to PARP inhibitors or experienced a recurrence of cancer.

[0022] In one embodiment of the present invention, the solid cancer may be, but is not limited to, ovarian cancer, breast cancer, prostate cancer, pancreatic cancer, colon cancer, gallbladder cancer, biliary tract cancer, or gastric cancer, which are known to be caused by BRCA1 / 2 mutations.

[0023] In one embodiment of the present invention, the solid cancer may be an advanced solid cancer, a recurrent solid cancer, or a metastatic solid cancer.

[0024] In one embodiment of the present invention, when the solid cancer is ovarian cancer, it may be advanced ovarian cancer, recurrent ovarian cancer, or high-grade serous ovarian cancer (including fallopian tube cancer or primary peritoneal cancer), and when the primary cancer is ovarian cancer and the metastatic cancer is ovarian cancer, it may be, but is not limited to, breast cancer, prostate cancer, pancreatic cancer, colorectal cancer, gallbladder cancer, biliary tract cancer, gastric cancer, liver cancer, or lung cancer.

[0025] In one embodiment of the present invention, the pharmaceutically acceptable salt of 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile may be a citrate salt.

[0026] The present invention also provides Provided is 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof for use in treating patients with solid tumors resistant to PARP inhibitors.

[0027] The present invention also provides Provided is a method for treating a patient with a solid cancer resistant to a PARP inhibitor by administering an effective amount of 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof.

[0028] The present invention also provides The present invention provides a use of 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof for the manufacture of a drug for treating patients with solid cancers resistant to PARP inhibitors. [Effects of the Invention]

[0029] The pharmaceutical composition according to the present invention comprising 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof can effectively reduce tumor size in patients with solid cancers resistant to PARP inhibitors, and is therefore useful for treating patients with solid cancers resistant to PARP inhibitors. [Brief explanation of the drawings]

[0030] [Figure 1] 1 is a graph showing the analysis results of the anti-cancer effect of the citrate compound of Chemical Formula 1 of the present invention (Example 2). [Figure 2] 1 is a graph showing the experimental results (Example 4) of the inhibition of Wnt signaling pathway activity by the citrate compound of Chemical Formula 1 of the present invention. [Figure 3] FIG. 1 shows the anti-cancer effect of the citrate compound of Chemical Formula 1 of the present invention evaluated using a Xenograft model (Example 5). [Figure 4] FIG. 1 shows the anti-cancer effect of the citrate compound of Chemical Formula 1 of the present invention evaluated using a Xenograft model (Example 6). DETAILED DESCRIPTION OF THE INVENTION

[0031] The present invention will now be described in further detail. In describing and claiming particular features of the present disclosure, the following terminology will be used in accordance with the definitions set out below unless otherwise indicated.

[0032] In this application, even if an embodiment is described with the term "comprising," it should be understood that other similar embodiments described with the terms "consisting of" and / or "consisting essentially of" are also provided.

[0033] The term "pharmaceutically acceptable" means a substance that is acceptable to patients from a pharmacological / toxicological standpoint in terms of composition, dosage form, safety, etc., and a "pharmaceutically acceptable carrier" refers to a medium that does not inhibit the effects of the biological activity of an active ingredient and is non-toxic to a subject when administered.

[0034] The term "resistance" refers to a case where there is no desired response (anti-cancer effect) to a drug. Specifically, in the present invention, it encompasses both a case where there is no response to the drug from the beginning despite standard therapy with a PARP inhibitor (refractory), and a case where there is a response to the drug at first but then recurs at a certain point (a case where the cancer lesion initially shrinks but then recurs and grows; acquired resistance). In this specification, "resistance" and "tolerance" can be used interchangeably.

[0035] The term "patient" or "subject" or "individual" refers to an organism suffering from a pathological condition, such as a solid tumor, that may be treated by administering a pharmaceutical composition of the present invention, and includes both humans and animals. Subjects include, but are not limited to, mammals (e.g., mice, monkeys, horses, cows, pigs, dogs, cats, etc.), preferably humans. Furthermore, "patient" or "subject" or "individual" in the present invention includes patients with solid tumors that are resistant to PARP inhibitors.

[0036] The term "BRCA1 / 2 mutation" refers to a mutation in BRCA1 and / or BRCA2, which refers to a naturally occurring mutation at one or more sites in the BRCA1 and BRCA2 genes. Thus, the mutation may occur in either one of the BRCA1 and BRCA2 genes, or in both genes, and the mutation may occur at one site or more sites in each gene.

[0037] As mentioned above, PARP inhibitors have shown great clinical potential as targeted therapeutic agents for homologous recombination-deficient tumors, but are known to have a high rate of innate or acquired resistance. There are various explanations for this, including i) increased drug efflux due to increased ABC transporters, ii) activation of PAR chains, iii) reactivation of the homologous recombination mechanism due to mutations in tumor suppressor genes such as p53 that affect the homologous recombination mechanism, iv) stabilization or protection of replication forks, and v) activation of the Wnt signaling pathway.

[0038] In relation to the mechanism of iii) above, it is known that various proteins are involved in the homologous recombination process, and in cancer patients in whom BRCA1 / 2 mutations have been found, mutations in other proteins involved in the homologous recombination process, such as p53, ATM, ATR, and p51, have also been found at the same time, and it has been explained that these are related to the acquisition of resistance to PARP inhibitors.

[0039] Although PARP inhibitors act as targeted therapeutic agents specific to patients with homologous recombination-deficient tumors, they are prone to developing resistance, and there is a growing demand for new anticancer drugs that can be used to treat patients resistant to PARP inhibitors.

[0040] Multidrug resistance (MDR), one of the causes of anticancer drug treatment failure, has recently emerged as a major issue in the field of anticancer drug therapy. This ability is due to the presence of the MDR gene in cancer cells. The MDR1 (ABCB1) gene produces a substance called P-glycoprotein (P-gp). P-gp is an enzyme that helps various drugs cross the cell membrane and efflux them from the cell interior to the extracellular space. Drug resistance develops when many anticancer drugs are administered to patients over a long period of time, and overexpression of P-gp is one of the mechanisms of resistance. Therefore, when P-gp is overexpressed, drugs that are P-gp substrates are effluxed from the cell by P-gp, rendering them ineffective. Multidrug resistance is a significant limiting factor in the use of anticancer drugs, and extensive research is being conducted to overcome this issue. Thus, cancer cells with overexpressed P-gp can overcome resistance by suppressing the function of P-gp or by selecting anticancer drugs that are not used as substrates for P-gp.

[0041] The citrate compound of Formula 1 of the present invention has been confirmed to have a significantly lower P-gp efflux rate than conventional PARP inhibitors, and is therefore useful for treating patients resistant to PARP inhibitors.

[0042] The Wnt signaling pathway is involved in embryonic development, tissue homeostasis, and various diseases. Hyperactivated signaling leads to the accumulation of β-catenin, which translocates into the nucleus and promotes oncogene transcription and cell growth. Therefore, efforts are being made to develop therapeutic agents that block the Wnt signaling pathway.

[0043] Recent studies have shown that the mechanism of resistance to PARP inhibitors is related to the Wnt signaling pathway. In other words, PARP inhibitors activate the Wnt signaling pathway, and it is known that resistance develops through activation of this Wnt signaling pathway. Therefore, by selecting anticancer drugs that can block the Wnt signaling pathway in cancer cells that have acquired resistance to PARP inhibitors, it is possible to overcome PARP inhibitor resistance.

[0044] The citrate compound of Formula 1 of the present invention has been confirmed to inhibit the Wnt signaling pathway, and is therefore useful for treating patients resistant to PARP inhibitors.

[0045] In the present invention, it has been found that when 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile (the compound of Chemical Formula 1) or a pharmaceutically acceptable salt thereof is administered to patients with solid tumors resistant to PARP inhibitors, the size of the solid tumors decreases.

[0046] This fact can be confirmed, for example, by applying the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof to a cell line resistant to a PARP inhibitor (olaparib, rucaparib, niraparib, or talazoparib) or primary cells (CHA-OVA-13 cells) isolated from a patient with cancer resistant to a PARP inhibitor (e.g., ovarian cancer). Specifically, the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof can be applied to a cell line resistant to a PARP inhibitor (e.g., IC50-13) among BRCA mutation-positive ovarian cancer or breast cancer cell lines such as HCC1937, SNU-251, BT474, and SNU-119. 50 After screening for cells with a cytotoxicity value of 50 μM or more, cell death can be confirmed by applying the compound of Formula 1 or a pharmaceutically acceptable salt thereof to the cell line.

[0047] Furthermore, the mechanism of anti-cancer activity of the compound of Chemical Formula 1 can be confirmed through experiments to confirm the expression levels of proteins related to cell death, homologous recombination, and signal transduction, such as pATR, pCHK1, pAKT, tankyrase, cleaved caspase 3, and cleaved PARP protein, in PARP inhibitor-resistant cell lines in which cell death occurs upon application of the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0048] This mechanism can also be confirmed by the results of animal models transplanted with PARP inhibitor-resistant cell lines and clinical trials involving actual PARP inhibitor-resistant cancer patients.

[0049] The present invention provides a pharmaceutical composition containing "6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile," a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof, for use in treating solid tumors in patients with resistance to PARP inhibitors.

[0050] <Chemical formula 1> TIFF2026026362000003.tif2664

[0051] The compound of Formula 1 exhibits inhibitory activity against PARP1 / 2, and therefore can be used as a targeted therapeutic agent for patients with homologous recombination-deficient tumors, and is also an anti-cancer agent that can simultaneously inhibit tankyrase1 / 2.

[0052] Tankyrase is involved in telomere homeostasis, Wnt / β-catenin signaling, glucose metabolism, and cell cycle progression. In particular, Wnt / β-catenin is involved in the transcriptional process of cancer-related genes. Because Wnt / β-catenin signaling is activated in various cancers, including gastrointestinal cancers, tankyrase inhibition has been reported to have anticancer effects by suppressing Wnt / β-catenin signaling. Indeed, attempts have been made to develop tankyrase inhibitors as anticancer drugs.

[0053] Therefore, the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof inhibits PARP1 / 2 like olaparib, which is used as a conventional standard treatment, and can also inhibit tankyrase, and therefore, it is understood that it acts through a mechanism different from that of olaparib, etc.

[0054] In the present invention, the pharmaceutically acceptable salt of Chemical Formula 1 is useful as an acid addition salt formed with a pharmaceutically acceptable free acid. The acid addition salt can be prepared by a conventional method, for example, by dissolving the compound in an excess of aqueous acid and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone, or acetonitrile. That is, the acid addition salt can be prepared by heating equal molar amounts of the compound and an acid or alcohol (e.g., glycol monomethyl ether) in water, followed by evaporating the solvent from the mixture to dryness, or by filtering the precipitated salt with suction.

[0055] In this case, the free acid may be an organic acid or an inorganic acid. The inorganic acid may be hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, or the like. The organic acid may be methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, or hydroiodic acid, but is not limited to these. In particular, the citrate salt of formula 1 may preferably be used.

[0056] In one embodiment of the present invention, the pharmaceutically acceptable salt of Chemical Formula 1 may be an anhydrate, monohydrate, or dihydrate of the citrate salt of Chemical Formula 1, and may be in a crystalline or amorphous form, or a mixture of the crystalline and amorphous forms.

[0057] Various forms of pharmaceutically acceptable salts of the compounds of Formula 1 may be prepared by methods known in the art.

[0058] In one embodiment of the present invention, examples of PARP inhibitors to which the solid cancer patient has resistance include, but are not limited to, anticancer drugs used in standard treatments, such as olaparib, rucaparib, niraparib, and talazoparib.

[0059] In one embodiment of the present invention, the PARP inhibitor may be olaparib. In one embodiment of the present invention, the solid cancer patient may be a patient with a homologous recombination deficient tumor having a BRCA1 / 2 mutation.

[0060] In one embodiment of the present invention, the BRCA1 / 2 mutation may be a germline mutation or a somatic mutation.

[0061] In one embodiment of the present invention, the patient may have a BRCA1 / 2 mutation and initially responded to a PARP inhibitor, but developed resistance during treatment and did not respond to the PARP inhibitor, or the cancer may have recurred.

[0062] In one embodiment of the present invention, the patient may be a patient who has a BRCA1 / 2 mutation but has not responded to a PARP inhibitor.

[0063] In one embodiment of the present invention, the patient may not have a BRCA1 / 2 mutation and may have previously failed to respond to PARP inhibitors or experienced a recurrence of cancer.

[0064] In one embodiment of the present invention, the solid cancer may be an advanced solid cancer, a recurrent solid cancer, or a metastatic solid cancer.

[0065] In one embodiment of the present invention, the solid cancer may be, but is not limited to, breast cancer, prostate cancer, pancreatic cancer, ovarian cancer, advanced ovarian cancer, high-grade serous ovarian cancer (including fallopian tube cancer or primary peritoneal cancer), metastatic cancer from primary ovarian cancer, breast cancer, prostate cancer, or pancreatic cancer.

[0066] In one embodiment of the present invention, the solid cancer may be ovarian cancer or a cancer that has metastasized from a primary ovarian cancer.

[0067] A pharmaceutical composition according to the invention may further comprise one or more pharmaceutically acceptable carriers or one or more excipients and / or diluents.

[0068] Examples of pharmaceutically acceptable carriers include, but are not limited to, solid and / or liquid phase substances such as ethanol, glycerol, water, etc. The amount of carrier in the pharmaceutical composition of the present invention may range from about 5 to about 99% by weight based on the total weight of the composition. Types of pharmaceutically acceptable excipients and diluents may include, but are not limited to, non-toxic compatible fillers, binders, disintegrants, buffers, preservatives, humectants, bulking agents, antioxidants, lubricants, flavoring agents, thickeners, coloring agents, surfactants, emulsifiers, suspending agents, etc. These excipients and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginic acid, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil, and it will be apparent to those skilled in the art that any other pharmaceutically acceptable carrier, excipient, and diluent may be used.

[0069] The pharmaceutical composition containing the compound of the present invention or a salt thereof may be formulated and used in the form of an oral dosage form such as a tablet, an acid, a granule, a pill, a capsule, a suspension, an emulsion, an oral solution, an emulsion, or a syrup, an external preparation, a suppository, or a sterile injectable solution, according to a conventional method.

[0070] The pharmaceutical compositions of the present invention may be in the form of a sterile injectable preparation as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated using suitable dispersing or wetting agents (e.g., Tween 80) and suspending agents according to techniques known in the art. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent (e.g., a solution in 1,3-butanediol). Acceptable vehicles and solvents include mannitol, water, Ringer's solution, or isotonic sodium chloride solution. Additionally, sterile, fixed oils may typically be used as solvents or suspending media. For this purpose, any non-irritating, fixed oil, including synthetic mono- or diglycerides, may be used. Fatty acids, such as oleic acid and its glyceride derivatives, are useful in injectable preparations, as are pharmaceutically acceptable natural oils (e.g., olive oil or castor oil), especially their polyoxyethylated versions.

[0071] Pharmaceutical compositions according to the present invention may be orally administered in any orally acceptable form including, but not limited to, capsules, tablets, and aqueous suspensions and solutions.

[0072] The pharmaceutical composition of the present invention for parenteral administration may be prepared in the form of suppositories or injections for rectal administration.For suppository compositions, the compound of the present invention may be mixed with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature.Such materials may include, but are not limited to, cocoa butter, beeswax, and polyethylene glycol.

[0073] In the case of an injectable composition, the compound of the present invention may be contained as an active ingredient in a conventional injectable vehicle, and the administration route may be, but is not limited to, intravenous injection, intramuscular injection, subcutaneous injection, etc.

[0074] The novel compound is contained in the pharmaceutical composition of the present invention in a therapeutically or prophylactically effective amount. The preferred dosage of the compound of the present invention varies depending on the patient's condition and weight, the severity of the disease, the drug form, the route of administration, and the duration of administration, and may be appropriately selected by those skilled in the art. However, to achieve the desired effect, the compound of Formula 1 of the present invention or a pharmaceutically acceptable salt thereof may be administered in an amount of 0.0001 to 1000 mg, 0.01 to 500 mg, 0.1 to 300 mg, 1 to 200 mg, or 50 to 200 mg once or several times a day. The compound of Formula 1 may be incorporated into the composition of the present invention in an amount of 0.0001 to 50 wt% based on the total weight of the entire composition.

[0075] The pharmaceutical composition of the present invention may further contain one or more active ingredients exhibiting the same or similar pharmacological effects in addition to the compound represented by Chemical Formula 1, its optical isomer, its racemate, or a pharmaceutically acceptable salt thereof.

[0076] The present invention also provides a use of the compound of Formula 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for preventing or treating solid cancer resistant to a PARP inhibitor.

[0077] The compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof for preparing a medicament may be mixed with a pharmaceutically acceptable adjuvant, diluent, carrier, etc., and prepared as a combined preparation together with other active ingredients to obtain a synergistic effect.

[0078] Furthermore, the present invention provides a method for preventing or treating solid cancer resistant to PARP inhibitors, by administering an effective amount of the compound of Formula 1 or a pharmaceutically acceptable salt thereof to a mammal, including a human.

[0079] The preventive or therapeutic method of the present invention includes not only treating a disease before symptoms appear but also inhibiting or avoiding the symptoms by administering the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof. In disease management, the preventive or therapeutic dose of a specific active ingredient varies depending on the nature and severity of the disease or condition and the route by which the active ingredient is administered. The dosage and frequency of administration vary depending on the age, weight, and response of each individual patient. Appropriate dosages can be easily selected by those skilled in the art, taking these factors into consideration. Furthermore, the preventive or therapeutic method of the present invention may further include administering a therapeutically effective amount of an additional active agent useful for treating the disease together with the compound represented by Chemical Formula 1, where the additional active agent provides a synergistic or additive effect with the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0080] The matters mentioned in the pharmaceutical composition, use and treatment method of the present invention are all applicable to the same unless they contradict each other. The pharmaceutical composition of the present invention may be provided in the form of a kit including instructions and the like.

[0081] Unless otherwise indicated, all numbers used in the specification and claims, whether stated or not, should be understood to be modified in all cases by the term "about." It should also be understood that the precise numerical values ​​used in the specification and claims form additional embodiments of the present disclosure. While efforts have been made to ensure the accuracy of the numerical values ​​disclosed in the examples, all measured numerical values ​​may inherently contain certain error values ​​resulting from the standard deviations observed in each measurement technique.

[0082] The present invention will be described in more detail below with reference to examples. These examples are merely for the purpose of explaining the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. [Example]

[0083] In the present invention, the citrate salt of Chemical Formula 1 may be prepared by a method known in the art or a method disclosed in Korean Patent Application No. 10-2021-0064416, or a method disclosed in an application filed on the same date as the present invention, which claims priority based on the above application. For example, a method for preparing the citrate salt of Chemical Formula 1 is as follows.

[0084] Manufacturing Example 1 Preparation of 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile citrate (citrate of formula 1)

[0085] Methanol (25.7 L) and purified water (25.7 L) were added to 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile (compound of Formula 1, 7.34 kg, 18.32 mol). Citric acid (5.28 kg, 27.49 mol) was dissolved in a 1:1 mixture of methanol and purified water (22 L) and then added. After stirring at 15-25°C for 30 minutes, the mixture was heated to 60°C and stirred at 60-70°C for 2 hours. After cooling to room temperature, the mixture was filtered to obtain the citrate monohydrate of Formula 1 (10.7 kg, 95.8%).

[0086] Ethanol (2.5 L), acetone (2.5 L), and isopropanol (2.5 L) were added to the citrate monohydrate of formula 1 (500 g, 0.82 mol), followed by the addition of purified water (20 mL). The mixture was heated to 55°C and stirred at 55-75°C for 4 hours. The mixture was cooled to below 25°C and stirred for 30 minutes. The resulting solid was filtered to obtain the anhydrous citrate of formula 1 (470 g, yield 96.7%).

[0087] Example 1: Analysis of the anti-cancer effect of Chemical Formula 1 (in vitro experiment) To analyze the anticancer effect of the compound of formula 1, 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile) citrate, in homologous recombination-deficient tumors, we performed mitotic cell analysis using various cell lines.

[0088] The cell lines used in this analysis were HCC1937, SNU-251, BT474, and SNU-119 as BRCA mutation-positive ovarian or breast cancer cell lines, and CHA-OVA-13 cells (primary ovarian cancer cells showing acquired resistance to olaparib) isolated from an actual BRCA1 mutation-positive ovarian cancer patient as primary BRCA mutation-positive ovarian cancer cells.

[0089] First, we performed IC to determine whether each cell line was resistant to olaparib. 50 At this time, IC for olaparib will be measured and confirmed. 50 Cell lines with a PARP inhibitor resistance of 50 uM or higher will be selected as PARP inhibitor-resistant cell lines suitable for this study.

[0090] The following experiment was performed on the olaparib-resistant cell line using various PARP inhibitors. Specifically, each cell was suspended in culture medium and distributed into a 96-well plate. After 24 hours of culture at 5% CO2 and 37°C, olaparib, niraparib, talazoparib, and the citrate salt of Formula 1 were dose-dependently administered. After 72 hours, MTT reagent was added, and after 3 hours, stop buffer (10% SDS) was added. After 2 to 4 hours of incubation, absorbance was measured at 595 nm, and the IC was determined as the concentration at which each drug inhibited cell growth by 50%. 50 Calculate the value.

[0091] At the same time, to confirm the cell death mechanism and anti-cancer activity mechanism of the cells administered each drug at the molecular level, the amount of proteins related to cell death, such as pATR, pCHK1, pAKT, tankyrase, cleaved caspase 3, and cleaved PARP protein, will be analyzed by immunoblotting.

[0092] Example 2: Analysis of anti-cancer effects (in vitro experiment) To analyze the anticancer effect of the compound of Chemical Formula 1, (6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile citrate, on BRCA wild-type and mutant-type cells, cell division assays were performed using various cell lines.

[0093] (1) Experimental method This assay used wild-type BRCA1 ovarian cancer cell lines OVCAR-3, OVCAR-5, SKOV3, NCI / ADR-RES, A2780, A2780 CR (a carboplatin-resistant cell line), and OVCA433R (an olaparib-resistant cell line), as well as the mutation-type BRCA1 ovarian cancer cell line SNU-251. The cell lines were cultured at 37°C in 5% CO2 in a culture medium (RPMI-1640 + 10% heat-inactivated FBS + 1% antibiotic-antimycotic). After culturing, the cells were removed from the cell culture dish and cultured overnight in a 6-well plate to allow cell attachment. The citrate compound of Formula 1, olaparib, and niraparib were serially diluted from the highest concentration to the lowest, and vehicle controls were used in the remaining wells. After 14 days, the cells were treated with crystal violet to stain live colonies. After washing with PBS and drying thoroughly at room temperature, the number of stained colonies was counted and recorded using a microscope and the naked eye. 50The value was calculated by converting the relative number of colonies in the experimental group into a percentage, assuming that the control group (vehicle control) was 100%, and determining the concentration at which colony formation could be inhibited by 50% (IC 50 The standard was an inhibitory concentration to achieve 50% colony formation inhibition.

[0094] (2) Experimental results The experimental results are shown in Table 1 below and FIG.

[0095] [Table 1]

[0096] As shown in Table 1 and the graph in Figure 1, the citrate compound of Formula 1 inhibits cancer cell growth at significantly lower concentrations than olaparib or niraparib, regardless of the presence or absence of BRCA mutations. In particular, it effectively inhibited cancer growth at significantly lower concentrations than other PARP inhibitors in the NCI / ADR-RES cell line, which exhibits drug resistance due to overexpression of drug efflux pumps, and the A2780-CR cell line, which has induced resistance to carboplatin.

[0097] Example 3: Evaluation of the P-gp efflux rate of the citrate compound of Formula 1 P-glycoprotein (P-gp) is one of the drug transporters that determines the absorption and efflux of various drugs. These drug absorption and efflux processes affect the drug concentrations in plasma and tissues and ultimately the final drug effects.

[0098] Compounds with high P-gp substrate specificity are known to cause reduced drug accumulation in multidrug-resistant cells and often mediate the development of resistance to anticancer drugs. PARP inhibitors such as olaparib, rucaparib, niraparib, and talazoparib are all known to have high P-gp substrate specificity.

[0099] In this experiment, the inventors attempted to clarify the mechanism by which the citrate compound of Chemical Formula 1 exhibits excellent efficacy in the treatment of cancers resistant to PARP inhibitors by comparing the efflux ratio of the citrate compound of Chemical Formula 1 with that of olaparib, a representative PARP inhibitor, in P-gp.

[0100] (1) Experimental method A permeability study was conducted to measure the efflux ratio of the citrate compound of Formula 1 and olaparib (AZD-2281). Five 10 CaCO2 cells were placed in each 6.5 mm diameter transwell insert in a 24-well plate. 4 200 μL of culture medium was dispensed into the apical side of the transwell inserts, corresponding to the number of cells per well. 800 μL of culture medium was dispensed into the basolateral side of the well plate, submerging the bottom of the inserts. After 21 days of culture, the culture medium was removed from the inserts and plates. The citrate compound of formula 1 was diluted in culture medium to concentrations of 1 μM, 10 μM, and 50 μM, respectively, and olaparib (AZD-2281) was diluted in culture medium to a concentration of 10 μM. 250 μL of each dilution was then applied to the apical side of the transwell inserts, and 800 μL of drug-free culture medium was added to the basal side (Papp A→B measurement). To examine the effects of the P-gp pump, 800 μL of culture medium containing the same drug concentration was applied to the basal side, and drug-free culture medium was added to the apical side of the inserts (Papp B→A measurement). At designated times, 0, 30, 60, 120, and 180 minutes, 100 μL samples were taken from either the superior or basolateral zones and analyzed.

[0101] (2) Experimental results The experimental results are shown in Table 2 below.

[0102] [Table 2]

[0103] Note) A: Upper layer of cell, B: Base of cell As can be seen from Table 2, the efflux rate of the citrate salt of Chemical Formula 1 was confirmed to be about 1 / 10 of that of olaparib, a PARP inhibitor that is a P-gp substrate.

[0104] (3) Evaluation of experimental results

[0105] The above experimental results indicate that the citrate compound of Formula 1 of the present invention is not a P-glycoprotein (P-gp) substrate.

[0106] Due to this mechanism of action, the citrate compound of Chemical Formula 1 is thought to be able to overcome multidrug resistance and exhibit superior anti-cancer effects, unlike PARP inhibitors such as olaparib, rucaparib, niraparib, and talazoparib, which are known to be P-glycoprotein (P-gp) substrates.

[0107] Specifically, in Example 2, the citrate compound of Chemical Formula 1 inhibited cancer cell growth in various wild-type and mutation-type BRCA ovarian cancer cell lines at significantly lower concentrations than olaparib and niraparib. This superior effect is believed to be partially attributable to the mechanism of action based on the efflux rate of P-gp. In particular, the results of Example 2 showed that in the NCI / ADR-RES cell line, which exhibits drug resistance due to overexpression of drug efflux pumps, the citrate compound of Chemical Formula 1 effectively inhibited cancer growth at significantly lower concentrations than other PARP inhibitors. Therefore, these experimental results are considered to more clearly explain that the fact that the compound of Chemical Formula 1 is not a P-gp substrate is partially responsible for its superior anti-cancer effect.

[0108] Furthermore, PARP inhibitors are known to have a high rate of congenital or acquired resistance, and one mechanism that may explain this is increased drug efflux due to an increase in ABC transporters.

[0109] Therefore, the mechanism of action of the citrate compound of Chemical Formula 1 on the efflux rate is believed to theoretically support the fact that the compound of Chemical Formula 1 can be effectively used to treat cancers resistant to other PARP inhibitors such as olaparib, rucaparib, niraparib, and talazoparib. This theory is clearly supported by the experimental results of Example 4 (Anti-cancer effect analysis - Xenograft model), Example 5 (Anti-cancer effect analysis - Xenograft model), and Example 6 (Phase 1 clinical trial - NOV140201) below.

[0110] In conclusion, the citrate compound of Formula 1 of the present invention has a different mechanism of action in terms of drug efflux mechanism from other PARP inhibitors such as olaparib, rucaparib, niraparib, and talazoparib, and provides excellent anti-cancer effects, even against cancers resistant to these PARP inhibitors.

[0111] Example 4: Evaluation of efficacy by inhibiting Wnt signaling activity associated with olaparib resistance (1) Experimental method A TOP / FOP-flash luciferase reporter assay was performed to measure the activation of Wnt signaling in the ovarian cancer cell line PEO1 and the olaparib-resistant ovarian cancer cell line PEO1-OR. The olaparib-resistant PEO1 cell line was treated with increasing concentrations of olaparib, starting from a low concentration (10 nM) to a higher concentration (8 μM). The cell line that survived was designated PEO1-OR. The TOP / FOP-flash luciferase reporter assay utilizes the principle that Wnt activation generates β-catenin, which translocates into the nucleus and binds to the TCF / LEF promoter, transcribing genes affected by Wnt. The TOP / FOP-flash luciferase reporter assay measures Wnt activation by measuring the luminescence produced by luciferase conversion of a non-luminescent substrate. TOP-flash is an experimental plasmid in which β-catenin binds to the TCF promoter, resulting in transcription of luciferase. FOP-flash is a transfection control plasmid in which a mutation is induced in the engineered TCF promoter, preventing β-catenin from binding to the promoter. It is used to measure the basal level of fluorescence.

[0112] The ovarian cancer cell line PEO1 and the olaparib-resistant ovarian cancer cell line PEO1-OR were transfected with TOP-flash or FOP-flash plasmids. For 72 hours after transfection, the transfected PEO1 and PEO1-OR cell lines were exposed to vehicle control and the citrate compound of Formula 1 at 400 nM, 10 μM, and 50 μM, respectively. The cells were then lysed and luciferase substrate was added. The degree of luminescence released from the substrate was measured and recorded using a fluorescence reader.

[0113] (2) Experimental results The results of the experiment are shown in FIG. As shown in the graph on the left of Figure 2, a relative comparison of the TOP signaling intensities between the ovarian cancer cell line PEO1 and the olaparib-resistant ovarian cancer cell line PEO1-OR showed that the olaparib-resistant PEO1-OR cell line exhibited TOP signaling intensity five times higher than that of the non-olaparib-resistant cell line. These results indicate that Wnt signaling was activated in the PEO1-OR cell line upon acquisition of olaparib resistance.

[0114] Furthermore, the graph on the right side of Figure 2 shows that when the Wnt-activated PEO1-OR cell line was treated with the citrate compound of Chemical Formula 1, Wnt signaling decreased in proportion to the concentration of the citrate compound of Chemical Formula 1 compared to the control group. In conclusion, these experiments demonstrate that when the PEO1 cell line acquires resistance to olaparib, Wnt signaling increases (see the graph on the left in Figure 2), and this increased signaling is reduced in a dose-proportional manner due to the tankyrase inhibitory ability of the citrate compound of Formula 1 (see the graph on the right in Figure 2).

[0115] (3) Evaluation of experimental results Recent studies have shown that the resistance mechanism to PARP inhibitors is related to the Wnt signaling pathway. In other words, it is known that the Wnt signaling pathway is activated as a mechanism for the development of resistance to PARP inhibitors. Therefore, by selecting anticancer drugs that can block the Wnt signaling pathway in cancer cells that have acquired resistance to PARP inhibitors, it is possible to overcome PARP inhibitor resistance.

[0116] Unlike conventional PARP inhibitors such as olaparib, the citrate compound of Chemical Formula 1 of the present invention has the characteristic of dually inhibiting PARP and tankyrase (TNK). That is, while conventional PARP inhibitors are known to activate the Wnt signaling pathway as resistance develops, the citrate compound of Chemical Formula 1 of the present invention effectively inhibits Wnt signaling through its TNK inhibitory activity.

[0117] Therefore, these facts indicate that the citrate compound of Formula 1 of the present invention can be effectively used in the treatment of cancers that have acquired resistance to PARP inhibitors.

[0118] Example 5: Analysis of anti-cancer effect (Xenograft model) To demonstrate that the citrate compound of Chemical Formula 1 can actually be used to treat solid tumors resistant to PARP inhibitors, a xenograft model was created using cells derived from BRCA mutation-positive ovarian cancer, and the effects of the PARP inhibitor (olaparib) and the citrate compound of Chemical Formula 1 were compared.

[0119] (1) Experimental method PDX-GTFP 1016 cells were derived from the primary tissue of a patient with stage IIIC high-grade serous ovarian cancer (PDX-1016 GTFP 1016 GFP / luc) and harbored TP53 and BRCA2 mutations. To facilitate cell tracking, green fluorescent protein / luciferase-expressing cells (PDX-1016 GTFP 1016 GFP / luc) were surgically injected into the right ovary intrabursally. Cancer cell establishment was monitored for 4 weeks prior to drug administration. After 4 weeks, olaparib-resistant PDX-1016 GTFP 1016 GFP / luc cells were defined as olaparib-resistant PDX-1016 GTFP 1016 GFP / luc (Ola-R-GTFP-1O16) cells.

[0120] Ola-R-GTFP-1O16 cells, an olaparib-resistant PDX established by the above method, were cultured at 1x10 6 The cells were surgically injected at a concentration of several cells / ml into the superior bursa of the right ovary, in the same location as the primary tumor, and allowed to stabilize for 4 weeks.

[0121] After 4 weeks of stabilization, image data such as the light intensity of GFP / luciferase expressed by the cancer was collected through in vivo flux imaging (IVIS spectrum in vivo imaging system, PerkinElmer), and the distribution and growth of the cancer were observed based on the flux (photons per second).

[0122] At this time, using in vivo flux imaging, transplanted mice with similar tumor growth rates were randomly divided into a control group (vehicle treatment group) and a treatment group with 25 mg / kg of the citrate compound of Chemical Formula 1, which were orally administered once daily, and changes in flux were recorded every week. After 4 weeks, the mice were sacrificed and the volume of ascites, the volume of cells in the ascites, and the number of cancer metastases to the lymph nodes were measured to determine the anti-cancer activity.

[0123] (2) Experimental results The experimental results are shown in FIGS. The strong spectral color of abdominal fluorescence in Figure 3B indicates active cancer growth in the ascites. In other words, Figure 3B shows that cancer growth was effectively suppressed in the citrate compound of Formula 1-treated group compared to the control group (vehicle-treated group).

[0124] Graph C of Figure 3 shows the results of in vivo imaging of the abdominal region of transplanted mice up to four weeks after drug treatment, with the in vivo fluorescence recorded from two weeks after drug treatment set as the basal level.

[0125] Graph C of FIG. 3 shows that the growth of cancer cells was significantly inhibited in the group treated with the citrate compound of Chemical Formula 1 compared to the control group (vehicle-treated group).

[0126] To confirm the correlation between treatment with the citrate compound of Chemical Formula 1 and the more effective inhibition of cancer growth compared to the control group, the number of cancer nodules and the number of cells in the ascites were measured, and the metastatic potential of the cancer was assessed.

[0127] As shown in Figure 3D, the nodule formation ability of the cancer was significantly reduced in the group treated with the citrate compound of Chemical Formula 1 compared to the control group. Also, as shown in Figure 3E, the total number of cells produced in the ascites was significantly reduced in the group treated with the citrate compound of Chemical Formula 1 compared to the control group.

[0128] These results confirm that the group treated with the citrate compound of Chemical Formula 1 suppresses cancer formation and metastasis more effectively than the control group.

[0129] Example 6: Analysis of anti-cancer effects (xenograft model) (1) Experimental method CHA-OVA-13 is a primary cell line established from ascites cells of an ovarian cancer patient with olaparib-acquired resistance, carrying a BRCA1 mutation. This CHA-OVA-13 was injected into NOD / SCID mice to allow the primary tumor to grow. When the tumor reached a certain size, it was excised, cut into small pieces, and then transplanted into the subcutaneous layer of nude mice to establish a mouse model. The tumor size was 80 mm. 3 Once tumor size reached 100%, mice bearing tumors of similar size were selected and randomly divided into an olaparib treatment group and a compound of Formula 1 citrate treatment group.

[0130] Thereafter, the olaparib treatment group was orally administered 50 mg / kg twice daily, and the citrate salt treatment group of the compound of Chemical Formula 1 was orally administered 50 mg / kg once daily. Tumor size was measured lengthwise and widthwise with a caliper every two days and recorded. Tumor volume was calculated using the following formula:

number

[0131] The olaparib-treated group and the citrate compound of Chemical Formula 1-treated group were observed for tumor size for 18 days. After 10 days of drug treatment, the olaparib-treated group was randomly divided into two groups. One group received olaparib continuously for 8 days (olaparib-treated group), while the other group received the citrate compound of Chemical Formula 1 orally at 50 mg / kg once daily for 8 days (olaparib-citrate compound of Chemical Formula 1 alternating treatment group), and the effect on tumor size was observed.

[0132] (2) Experimental results The experimental results are shown in Figure 4. As shown in the graph of Figure 4(b), it can be seen that, over a total of 18 days of observation, tumor formation in the group treated with the citrate compound of Chemical Formula 1 was significantly delayed compared to the group treated with olaparib.

[0133] In addition, in the olaparib-citrate compound of Chemical Formula 1 alternating treatment group, in which olaparib was replaced with the citrate compound of Chemical Formula 1 at day 10, the tumor growth rate gradually slowed compared to the olaparib-only treatment group, and from around day 13, it showed a tumor growth inhibitory effect similar to that of the citrate compound of Chemical Formula 1-only treatment group.

[0134] These results indicate that continued treatment of primary cell xenografts from patients resistant to olaparib with olaparib does not inhibit tumor growth, but treatment with the citrate compound of Formula 1 significantly slows tumor growth.

[0135] That is, these results confirm that the citrate compound of Chemical Formula 1 has an excellent effect in inhibiting the growth of olaparib-resistant cancer tissues.

[0136] Example 7: Phase 1 clinical trial A Phase 1 clinical trial was conducted to evaluate the stability, drug resistance, pharmacokinetic and pharmacodynamic properties, and efficacy of the citrate salt of 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile, a compound of formula 1. Patients were aged 19 years or older with histologically or cytologically confirmed advanced solid cancer who were refractory to or unable to undergo standard treatment. Patients were selected based on an expected survival time of 12 weeks or longer and had normal hematological and liver function confirmed by routine blood tests.

[0137] Selected patients provided written consent in accordance with the study institution and Ministry of Food and Drug Safety regulations. Twenty-two patients were enrolled in the dose-escalation cohort, and 40 patients were enrolled in the dose-expansion cohort (6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile citrate salt). The dose-expansion cohort received 50 mg / day, 100 mg / day, 150 mg / day, and 200 mg / day.

[0138] All patients in the cohort underwent safety assessments, periodic blood sampling, biomarker analysis (tumor samples) by biopsy, genetic analysis (PBMC and tumor samples), and tumor response evaluation (every 6 weeks) according to regulations. Patients were also followed up after the end of treatment in accordance with the regulations of the testing institution and the Ministry of Food and Drug Safety.

[0139] Among the patients enrolled in the dose expansion cohort (150 mg / day), a 64-year-old female patient was initially diagnosed with metastatic ovarian cancer (stage 3). She had undergone extensive surgical resection of solid tumor tissue, including the ovaries, and had a homologous recombination-deficient tumor with a confirmed germline BRCA1 gene mutation.

[0140] The patient showed little response to various anticancer drugs, including gemcitabine, cisplatin, and paclitaxel. Immediately after administration of olaparib, the patient showed an increase in the size of the target lesion and new progressive disease. Resistance to olaparib was confirmed. Neoplatin, a cisplatin derivative, was administered for two months, but this was unsuccessful. The patient then enrolled in this clinical trial and began taking 150 mg / day of Formula 1 citrate alone. Periodic CT scans during treatment confirmed that the lesion size (solid tumor metastasized to the liver) had decreased by more than 30% compared to baseline.

[0141] Example 8: Phase 2 clinical trial A Phase 2 clinical trial will be conducted to evaluate whether the citrate salt of the compound of Chemical Formula 1 (6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile) can be used to treat patients with resistance to PARP inhibitors. Patients will be aged 19 years or older with histologically or cytologically confirmed solid tumors whose tumors have been confirmed to be homologous recombination deficiency (HRD). Specifically, patients with high-grade (Grade 2 or 3) serous epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer who have undergone two or more lines of anticancer drug therapy for their tumors prior to participating in this clinical trial and whose tumors have recurred or progressed.

[0142] The anticancer drug treatment specifically means treatment with any one or a combination of gemcitabine, doxorubicin, topotecan, carboplatin, oxaliplatin, cisplatin, bevacizumab, PARP inhibitors, and the like.

[0143] Patients will be selected as subjects whose expected survival period is 12 weeks or longer and whose hematological, renal and liver functions have been confirmed to be normal through routine blood tests.

[0144] Selected patients will provide written consent in accordance with the regulations of the study institution and the Ministry of Food and Drug Safety. Patients include those who have shown sensitivity to platinum-based therapeutic agents in their previous treatment history, or those who have shown sensitivity to platinum-based therapeutic agents but have failed treatment with conventional PARP inhibitors such as olaparib and niraparib. Approximately 60 patients will be enrolled, but this is subject to change.

[0145] The dosage of the citrate salt of Chemical Formula 1 is 100 mg / day, and the dosage can be adjusted as needed.

[0146] All cohort patients will undergo safety assessment, periodic blood sampling, genetic analysis (PBMC and tumor samples), and tumor response assessment (every 8 weeks) according to regulations. Patients will also be followed up after the end of treatment in accordance with regulations of the testing institution and the Ministry of Food and Drug Safety.

[0147] This phase 2 clinical trial will enable us to measure and evaluate efficacy, safety, drug resistance, PK, etc.

[0148] The efficacy includes the therapeutic effect of the "citrate salt of Chemical Formula 1 of the present invention" on "patients with HRD mutation-positive tumors who have failed treatment with conventional PARP inhibitors, etc." More specifically, this Phase 2 clinical trial will show that the size of solid tumors can be effectively reduced in patients who have failed treatment with conventional PARP inhibitors, etc.

Claims

1. A pharmaceutical composition for treating or preventing cancer in a patient with a solid cancer that is resistant to a PARP inhibitor, the pharmaceutical composition comprising 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof, wherein the solid cancer does not have resistance to 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition of claim 1, wherein the solid cancer patient has a homologous recombination deficiency (HRD) tumor.

3. The pharmaceutical composition of claim 2, wherein the solid cancer patient has a BRCA1 / 2 mutation.

4. The pharmaceutical composition of claim 3, wherein the BRCA1 / 2 mutation is a germline mutation.

5. The pharmaceutical composition of claim 3, wherein the BRCA1 / 2 mutation is a somatic mutation.

6. The pharmaceutical composition of claim 1, wherein the solid cancer patient does not have a BRCA1 / 2 mutation.

7. 2. The pharmaceutical composition of claim 1, wherein the PARP inhibitor is one or more selected from olaparib, rucaparib, niraparib, and talazoparib.

8. 8. The pharmaceutical composition of claim 7, wherein the PARP inhibitor is olaparib.

9. The pharmaceutical composition according to claim 1, wherein the solid cancer is one or more selected from breast cancer, prostate cancer, pancreatic cancer, ovarian cancer, advanced ovarian cancer, high-grade serous ovarian cancer (including fallopian tube cancer or primary peritoneal cancer), and metastatic cancer that has metastasized from primary ovarian cancer.

10. 10. The pharmaceutical composition of claim 9, wherein the solid cancer is ovarian cancer.

11. The pharmaceutical composition according to claim 9, wherein the solid cancer is a metastatic cancer that has metastasized from a primary cancer, ovarian cancer.

12. 12. The pharmaceutical composition of any one of claims 1 to 11, wherein the pharmaceutically acceptable salt of 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile is a citrate salt.

13. The pharmaceutical composition of any one of claims 1 to 11, further comprising a pharmaceutically acceptable carrier or excipient.

14. Use of 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating a patient with a solid tumor resistant to a PARP inhibitor, wherein the solid tumor is not resistant to 6-{4-[(5-oxo-1,2,3,4,5,6-hexahydrobenzo[h][1,6]naphthyridin-8-yl)methyl]piperazin-1-yl}nicotinonitrile or a pharmaceutically acceptable salt thereof.

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