Combination drug containing liposome composition containing topotecan or salt thereof and DNA damage repair inhibitor

Abstract

The present invention addresses the problem of providing a drug obtained by combining a liposome composition containing topotecan or a salt thereof and a DNA damage repair inhibitor. According to the present invention, provided is a drug which comprises (A) a liposome composition containing an aqueous solution having an inner aqueous phase and an outer aqueous phase in which the liposome is dispersed, and (B) a DNA damage repair inhibitor, and which is simultaneously or successively administered the liposome composition and the DNA damage repair inhibitor, the liposome composition (A) containing an aqueous solution having an inner aqueous phase and an outer aqueous phase in which the outer aqueous phase is dispersed, and the DNA damage repair inhibitor (B) containing a DNA damage repair inhibitor. Wherein the liposome contains topotecan or a salt thereof, and the lipid forming the liposome contains dihydrosphingomyelin.

Description

Technical Field

[0001] The present invention relates to a drug that combines a liposomal composition containing topotecan or a salt thereof and a DNA damage repair inhibitor and administers them simultaneously or sequentially. Background Art

[0002] In chemotherapy, a therapy has been widely studied in which a drug is accumulated in a lesion site such as cancer by a liposomal composition and is exposed thereto for a long time.

[0003] Non-Patent Document 1 describes the combined use of a topoisomerase I inhibitor and a DNA damage repair inhibitor.

[0004] Patent Document 1 describes the combined use of topoisomerase I (irinotecan) contained in liposomes and a PARP inhibitor as a DNA damage repair inhibitor.

[0005] Patent Document 2 describes a liposomal composition containing topotecan or a salt thereof. Patent Document 3 describes the combined use of a liposomal composition containing topotecan or a salt thereof and an immune checkpoint inhibitor. Patent Document 4 describes the combined use of a liposomal composition containing topotecan or a salt thereof and a platinum preparation.

[0006] Prior Art Documents

[0007] Patent Documents

[0008] Patent Document 1: Japanese Patent Application Laid-Open No. 2018-528184

[0009] Patent Document 2: International Publication No. 2018 / 181963

[0010] Patent Document 3: International Publication No. 2019 / 244979

[0011] Patent Document 4: International Publication No. 2020 / 071349

[0012] Non-Patent Documents

[0013] Non-Patent Document 1: CLINICAL CANCER RESEARCH, Vol. 25, No. 22, pp. 6581 - 6589 (2019): Targeting Topoisomerase I in the Era of Precision Medicine. Anish Thomas et al. Summary of the Invention

[0014] Technical Problem to be Solved by the Invention

[0015] Non-Patent Document 1 described above discloses the combined use of topoisomerase I and a DNA damage repair inhibitor. However, when these are combined, although a strong anti-tumor effect can be obtained, normal cells including highly proliferative hematopoietic stem cells are also severely damaged, so the safety is insufficient and further improvement is needed.

[0016] Patent Document 1 described above discloses the combined use of a liposome containing irinotecan as a topoisomerase I inhibitor and a PARP inhibitor. However, irinotecan is a prodrug (a precursor activated by an enzyme in the body), and its drug effects vary. Moreover, when it is incorporated into a liposome, the anti-tumor effect is weak, and side effects such as diarrhea are a concern, so further improvement is needed.

[0017] An object of the present invention is to provide a combination of two or more anti-cancer agents with high therapeutic effects and few side effects by combining two or more anti-cancer agents that act through different mechanisms when a liposome composition containing topotecan or a salt thereof and a DNA damage repair inhibitor are used in combination.

[0018] Means for Solving the Technical Problem

[0019] The present inventors conducted intensive studies to solve the above problems, and as a result, found that a drug obtained by combining (A) a liposome composition and (B) a DNA damage repair inhibitor and administering them simultaneously or sequentially can solve the above problems, thus completing the present invention. The (A) liposome composition contains liposomes having an inner aqueous phase and an aqueous solution as an outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipid constituting the liposomes contains dihydrosphingomyelin.

[0020] That is, the present invention provides the following.

[0021] [1] A drug comprising a combination of (A) a liposome composition and (B) a DNA damage repair inhibitor and administering the liposome composition and the DNA damage repair inhibitor simultaneously or sequentially, wherein the (A) liposome composition contains liposomes having an inner aqueous phase and an aqueous solution as an outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipid constituting the liposomes contains dihydrosphingomyelin.

[0022] [2] The drug according to [1], wherein

[0023] after administering the (A) liposome composition, the (B) DNA damage repair inhibitor is administered.

[0024] [3] The drug according to [1], wherein

[0025] After administering (B) a DNA damage repair inhibitor, (A) the liposome composition is administered.

[0026] [4] The medicament according to any one of [1] to [3], wherein,

[0027] The DNA damage repair inhibitor is at least one selected from PARP inhibitors, ATR inhibitors, ATM inhibitors, CHK1 / 2 inhibitors, WEE1 inhibitors, DNA-PK inhibitors, and inhibitors that inhibit the pathways related to them.

[0028] [5] The medicament according to any one of [1] to [4], wherein,

[0029] The lipid constituting the liposome further includes cholesterol and a lipid modified with polyethylene glycol.

[0030] [6] The medicament according to any one of [1] to [5], wherein,

[0031] The blending ratio of the lipid modified with polyethylene glycol is 2 mol% to 10 mol% relative to the total lipids constituting the liposome.

[0032] [7] The medicament according to any one of [1] to [6], wherein,

[0033] The lipid modified with polyethylene glycol is diacyl phosphatidylethanolamine modified with polyethylene glycol or methoxypolyethylene glycol.

[0034] [8] The medicament according to any one of [1] to [7], wherein,

[0035] The blending ratio of cholesterol is 35 to 43 mol% relative to the total lipids constituting the liposome.

[0036] [9] The medicament according to any one of [1] to [8], wherein,

[0037] The inner aqueous phase of the liposome contains an ammonium salt.

[0038]

[10] The medicament according to any one of [1] to [9], wherein,

[0039] The DNA damage repair inhibitor is a PARP inhibitor.

[0040]

[11] The medicament according to any one of [1] to

[10] , wherein,

[0041] (A) The dosage of topotecan or its salt contained in the liposome composition per administration is 0.1 mg / m 2 body surface area to 10 mg / m 2 body surface area.

[0042]

[12] The drug according to any one of [1] to

[11] , wherein,

[0043] (A) The liposomal composition is administered once every 1 to 8 weeks.

[0044]

[13] A treatment method, which is a treatment method for a target disease (preferably cancer), wherein,

[0045] At an effective dosage and administration period that show a synergistic effect in treatment, (A) the liposomal composition and (B) a DNA damage repair inhibitor are combined and administered simultaneously or sequentially, the (A) liposomal composition contains liposomes having an inner aqueous phase and an aqueous solution as the outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipid constituting the liposomes contains dihydrosphingomyelin.

[0046]

[14] A maintenance therapy, which is a maintenance therapy for a target disease (preferably cancer), wherein,

[0047] At an effective dosage and administration period required to prevent recurrence of the target disease, (A) the liposomal composition and (B) a DNA damage repair inhibitor are combined and administered simultaneously or sequentially, the (A) liposomal composition contains liposomes having an inner aqueous phase and an aqueous solution as the outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipid constituting the liposomes contains dihydrosphingomyelin.

[0048]

[15] A drug, which is used in the treatment of a target disease (preferably cancer), and its combination contains (A) the liposomal composition and (B) a DNA damage repair inhibitor, and the above liposomal composition and DNA damage repair inhibitor are administered simultaneously or sequentially, the (A) liposomal composition contains liposomes having an inner aqueous phase and an aqueous solution as the outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipid constituting the liposomes contains dihydrosphingomyelin.

[0049]

[16] Use of (A) a liposomal composition and (B) a DNA damage repair inhibitor for manufacturing a drug whose combination contains (A) the liposomal composition and (B) the DNA damage repair inhibitor and the above liposomal composition and DNA damage repair inhibitor are administered simultaneously or sequentially, wherein,

[0050] The (A) liposomal composition contains liposomes having an inner aqueous phase and an aqueous solution as the outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipid constituting the liposomes contains dihydrosphingomyelin.

[0051] Advantages of the Invention

[0052] The drug of the present invention combines a liposomal composition containing topotecan or its salt and a DNA damage repair inhibitor and administers them simultaneously or sequentially, which can reduce the enhancement of side effects in normal tissues (normal organs, bone marrow, etc.) and can provide strong drug efficacy against cancer. Therefore, it is possible to provide a drug that enhances at least one of the effects of treating or preventing cancer and suppressing recurrence.

[0053] Moreover, even at a low dosage, the drug of the present invention has a significant cell proliferation inhibitory effect in tissues. Therefore, for subjects including patients, it is possible to perform an ideal treatment that is not only highly safe, has a low physical burden, but also has high convenience. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 Shows the change in tumor volume from the administration date as data on the drug efficacy of Test Example 1.

[0055] Figure 2 Shows the change in body weight from the administration date as data on the safety of Test Example 1.

[0056] Figure 3 Shows the change in tumor volume from the administration date as data on the drug efficacy of Test Example 2.

[0057] Figure 4 Shows the change in body weight from the administration date as data on the safety of Test Example 2.

[0058] Figure 5 Shows the results of co - administration of Ceralasertib in Capan - 1 (pancreatic cancer cell line) for the isobologram analysis of Test Example 3.

[0059] Figure 6 Shows the results of co - administration of M4076 in Capan - 1 (pancreatic cancer cell line) for the isobologram analysis of Test Example 3.

[0060] Figure 7 Shows the results of co - administration of M3814 in Capan - 1 (pancreatic cancer cell line) for the isobologram analysis of Test Example 3.

[0061] Figure 8 Shows the results of co - administration of Zn - C3 in Capan - 1 (pancreatic cancer cell line) for the isobologram analysis of Test Example 3.

[0062] Figure 9 Shows the results of co - administration of Olaparib in Capan - 1 (pancreatic cancer cell line) for the isobologram analysis of Test Example 3.

[0063] Figure 10 Shows the results of combination with Ceralasertib in ES-2 (ovarian cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0064] Figure 11 Shows the results of combination with M4076 in ES-2 (ovarian cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0065] Figure 12 Shows the results of combination with Prexasertib in ES-2 (ovarian cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0066] Figure 13 Shows the results of combination with M3814 in ES-2 (ovarian cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0067] Figure 14 Shows the results of combination with Zn-C3 in ES-2 (ovarian cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0068] Figure 15 Shows the results of combination with Olaparib in ES-2 (ovarian cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0069] Figure 16 Shows the results of combination with Ceralasertib in DMS114 (lung cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0070] Figure 17 Shows the results of combination with M4076 in DMS114 (lung cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0071] Figure 18 Shows the results of combination with Prexasertib in DMS114 (lung cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0072] Figure 19 Shows the results of combination with M3814 in DMS114 (lung cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0073] Figure 20 Shows the results of combination with Zn-C3 in DMS114 (lung cancer cell line) for the equivalent line graph analysis related to Test Example 3.

[0074] Figure 21 Shows the results of combination with Olaparib in DMS114 (lung cancer cell line) for the equivalent line graph analysis related to Test Example 3. Detailed implementation mode

[0075] In the present invention, unless otherwise specified, the range indicated by "~" includes the values at both ends.

[0076] The subjects include humans and mammals other than humans. Examples of mammals other than humans include monkeys, dogs, cats, cows, horses, mice, rats, etc.

[0077] The treatment can be any treatment and therapy that can achieve the desired therapeutic effect (e.g., inhibiting or delaying the progression of the condition), including slowing down the progression rate, interrupting the progression rate, improvement of the condition, cure or remission of the condition (whether partial remission or complete remission), prevention, delay, alleviation or cessation of one or more symptoms and / or signs in the condition, or prolongation of the survival of the subject or the survival predicted for the subject compared to the case without treatment.

[0078] The treatment also includes prevention. For example, by treating a subject who is prone to developing or relapsing cancer or at risk of developing or relapsing cancer, the onset or recurrence of cancer in the subject can be prevented or delayed.

[0079] The treatment can include inhibition of cancer growth and / or inhibition of cancer metastasis (including complete remission of cancer). The growth of cancer refers to the transformation of cancer into a more developed form. As indicators for measuring the inhibition of cancer growth, examples include a decrease in the survival of cancer cells, a decrease in the tumor volume or morphology (e.g., determined using computed tomography (CT), ultrasonography or other imaging diagnostic methods), a delay in tumor growth, disruption of the tumor vasculature, improvement in the results of a delayed hypersensitivity skin test, an increase in the activity of cytolytic T-lymphocytes, and a decrease in the level of tumor-specific antigens, etc.

[0080] In the present invention, tumors, malignancies, cancers, malignant neoplasms, carcinomas, sarcomas, etc. are collectively referred to as "tumors" or "cancers". And, "tumors" or "cancers" include those that relapse after cancer treatment. "Tumors" include all neoplastic cell growth and proliferation, whether malignant or benign, as well as pre-cancerous and cancerous cells and tissues.

[0081] "Effective amount" is the dosage required to achieve the desired therapeutic or prophylactic result, including the time and amount of administration. The "effective amount" of the drug of the present invention can vary depending on the disease state, age, sex and weight of the subject (or individual), and the ability of the drug to elicit the desired response in the subject (or individual), etc.

[0082] "Concurrent administration" means administering the first therapy and the second therapy in a combination therapy at a time interval of about 15 minutes or less, such as about 10 minutes, about 5 minutes, or about 1 minute or less. When the first therapy and the second therapy are concurrently administered, the first therapy and the second therapy can be included in the same composition (for example, a composition containing both the first therapy and the second therapy), or can be included in different compositions (for example, the first therapy is included in one composition, and the second therapy is included in another composition).

[0083] The term "sequential administration" means administering the first therapy and the second therapy in a combination therapy at a time interval of more than about 15 minutes, such as about 20 minutes, about 30 minutes, about 40 minutes, about 50 minutes, about 60 minutes, or a longer time (1 day, 2 days, 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 8 weeks, 12 weeks, 16 weeks, etc.). In the present invention, in sequential administration, the first therapy can be administered first, and the case of administering the second therapy first is also included. And, in the present invention, sequential administration also includes the case of administering the second therapy after the first therapy (after a specific time (for example, 1 week later)). The first therapy and the second therapy can be included in different compositions, and these can be included in the same package or kit, or can be included in different packages or kits.

[0084] "Maintenance therapy" is a treatment method in which, after obtaining a certain effect through cancer surgery or drug therapy, the same or different drugs are continuously administered as much as possible for the purpose of preventing cancer recurrence or progression.

[0085] "Blood retention" means the property that a drug in a state encapsulated in liposomes exists in the blood in a subject to whom a liposome composition has been administered.

[0086] Unless otherwise specifically stated, the "average particle size of liposomes" means the average particle size measured by dynamic light scattering method (preferably the cumulant average particle size). As commercially available measuring devices for using dynamic light scattering, examples include a high-concentration system particle size analyzer FPAR-1000 (manufactured by Otsuka Electronics Co., Ltd.), NANOTRAC UPA (manufactured by NIKKISO CO., LTD.), and Nanosizer (manufactured by Malvern). It is also possible to calculate the volume average particle size or number average particle size of liposomes using conversion formulas inherent to the measuring devices of each manufacturer. When measuring particles of about 100 nm, it is not possible to accurately grasp the particle distribution by static light scattering method, etc., so it is preferable to use the dynamic light scattering method for measurement.

[0087] Hereinafter, the present invention will be described in detail.

[0088] The present invention relates to a medicament, the combination of which contains (A) a liposome composition and (B) a DNA damage repair inhibitor, and the liposome composition and the DNA damage repair inhibitor are administered simultaneously or sequentially. The (A) liposome composition contains liposomes having an inner aqueous phase and an aqueous solution as an outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipids constituting the liposomes contain dihydrosphingomyelin.

[0089] In Patent Document 2 described above, a liposome composition containing topotecan or a salt thereof is described, but the combination with a DNA damage repair inhibitor is not described. Therefore, it is not easy to conceive the medicament of the present invention from the description of Patent Document 2.

[0090] In Patent Document 3 described above, the combined use of a liposome composition containing topotecan or a salt thereof and an immune checkpoint inhibitor is described. By the action of the liposome containing topotecan or a salt thereof, the population such as the number and type of immune cells in the tumor environment changes. For the immune cells in the tumor environment, the T cells or antigen-presenting cells on which the immune checkpoint inhibitor acts enhance the anti-tumor effect, and thus the anti-tumor effect is exerted. This is a technique having an anti-tumor effect produced by an indirect synergistic effect via T cells or antigen-presenting cells.

[0091] On the other hand, the combination of a liposome composition containing topotecan or a salt thereof and a DNA damage repair inhibitor has a direct synergistic effect of damaging the gene (DNA) of cancer cells and inhibiting the repair of the damage. Therefore, it is not easy to conceive the medicament of the present invention from the description of Patent Document 3.

[0092] In Patent Document 4 described above, the combined use of a liposome composition containing topotecan or a salt thereof and a platinum preparation is described. Both the liposome composition containing topotecan or a salt thereof and the platinum preparation damage the gene (DNA) of cancer cells. Since the anti-tumor effect is obtained by a similar mechanism, it is not easy to conceive the medicament of the present invention from the description of Patent Document 4.

[0093] Moreover, in cancers (tumors) having tolerance to DNA damage, the effect may be reduced by the combination of a liposome composition containing topotecan or a salt thereof and a platinum preparation.

[0094] (Liposome)

[0095] A liposome is a closed small vesicle formed by a lipid bilayer membrane using lipids, and has an aqueous phase (inner aqueous phase) in the space of its closed vesicle. The inner aqueous phase includes water and the like. Liposomes usually exist in a state dispersed in an aqueous solution (outer aqueous phase) outside the closed vesicle. Liposomes can be single lamella (also called single-layer lamella or uni-lamella, with a structure where the bilayer membrane is a single layer), or multi-layer (also called multi-lamella, with a structure of multiple bilayer membranes in an onion-like shape. Each layer is separated by an aqueous layer), but in the present invention, from the viewpoints of safety and stability for drug use, single-layer liposomes are preferred.

[0096] As long as the liposome can contain a drug, its morphology is not particularly limited. "Containing" means a form in which the drug is contained in the inner aqueous phase of the liposome. For example, forms such as encapsulating the drug into a closed space formed by a membrane, containing it within the membrane itself, etc. can be cited, and these can also be combined.

[0097] The average particle size of liposomes is usually 10 nm to 1000 nm, preferably 20 nm to 500 nm, more preferably 30 nm to 300 nm, further preferably 30 nm to 200 nm, still further preferably 30 nm to 150 nm, and particularly preferably 50 nm to 150 nm. Liposomes are preferably spherical or a shape similar thereto.

[0098] In the case where the enhanced permeability and retention (EPR) effect is expected, the substantial diameter is preferably 50 to 200 nm, more preferably 50 to 150 nm, and further preferably 50 to 100 nm. The term "substantially" means that at least 75% of the number of liposomes is within the specified diameter range. Regarding the said "at least 75%", it is more preferably at least 80%, and further preferably at least 90%.

[0099] In addition, regarding the average particle size of liposomes, in the present invention, unless otherwise specifically stated, the "average particle size" refers to the average particle size measured by dynamic light scattering method (preferably the cumulative average particle size). The "average particle size" can be measured by using a device capable of measuring the average particle size by light scattering method.

[0100] The components of the lipid bilayer constituting the liposome are selected from lipids. The liposomes in the present invention preferably contain diacyl phosphatidylethanolamine modified with a hydrophilic polymer, dihydrosphingomyelin, and cholesterol as the components constituting the liposome membrane.

[0101] The liposome of the present invention contains dihydrosphingomyelin. By using dihydrosphingomyelin, the retention of the liposome in the blood can be improved. Also, the barrier property of the liposome membrane can be enhanced, thereby preventing the leakage of topotecan or its salt.

[0102] Dihydrosphingomyelin usually has two long-chain alkyl groups in the molecule. Examples of dihydrosphingomyelin include those having two long-chain alkyl groups with 16 carbon atoms, those having a long-chain alkyl group with 16 carbon atoms and a long-chain alkyl group with 18 carbon atoms, and those having a long-chain alkyl group with 16 carbon atoms and a long-chain alkyl group with 20 - 24 carbon atoms.

[0103] As dihydrosphingomyelin, from the viewpoint of preventing the leakage of drugs from liposomes, it is preferable to use the following compound having a long-chain alkyl group with 16 carbon atoms and a long-chain alkyl group with 18 carbon atoms. The reason is that the higher the number of carbon atoms, the higher the melting point, and a liposome membrane with high barrier property can be produced.

[0104] [Chemical formula 1]

[0105]

[0106] As dihydrosphingomyelin, for example, dihydrosphingomyelin obtained by reducing sphingomyelin derived from natural substances by a conventional method can be used, or dihydrosphingomyelin obtained by synthesis can be used.

[0107] Since dihydrosphingomyelin derived from natural substances such as eggs mostly has two long-chain alkyl groups with 16 carbon atoms, from the viewpoint of obtaining dihydrosphingomyelin having a long-chain alkyl group with 16 carbon atoms and a long-chain alkyl group with 18 carbon atoms in high purity, it is preferable to use dihydrosphingomyelin obtained by chemical synthesis.

[0108] The ratio of dihydrosphingomyelin in all the lipids constituting the liposome is preferably 30 - 80 mol%, more preferably 40 - 70 mol%, and further preferably 50 - 60 mol%.

[0109] The liposome of the present invention preferably contains a lipid modified with polyethylene glycol (hereinafter referred to as PEG-modified lipid). In addition, derivatives of polyethylene glycol can also be used. Examples of derivatives include methoxypolyethylene glycol, etc. Hereinafter, when referred to as PEG modification, it also includes derivatives of polyethylene glycol.

[0110] Examples of PEG-modified lipids include PEG-modified phospholipids, PEG-modified monoglycerides, PEG-modified diglycerides, PEG-modified sorbitan fatty acid esters, PEG-modified monoalkyl ethers, PEG-modified sterols, etc. The above hydrophilic polymers can be used alone or in combination of two or more.

[0111] The molecular weight of polyethylene glycol is not particularly limited and is 500 to 10,000 daltons, preferably 1,000 to 7,000 daltons, and more preferably 2,000 to 5,000 daltons.

[0112] Examples of the PEG-modified lipid include 1,2-distearoyl-3-phosphatidylethanolamine-PEG2000 (manufactured by NOF CORPORATION), distearoyl glycerol-PEG2000 (manufactured by Nippon Oil&Fats Co., Ltd.), 1,2-distearoyl-3-phosphatidylethanolamine-PEG5000 (manufactured by Nippon Oil&Fats Co., Ltd.), etc., such as 1,2-distearoyl-3-phosphatidylethanolamine-polyethylene glycol, cholesterol-PEG 600 (manufactured by Merck.Ltd), etc. cholesterol-polyethylene glycol.

[0113] From the viewpoints of versatility and blood retention, as the PEG-modified lipid, PEG-modified phospholipids and PEG-modified monoalkyl ethers are preferred, and PEG-modified phospholipids are more preferred.

[0114] Among the PEG-modified phospholipids, PEG-modified phosphatidylethanolamine is preferred, which is a diacyl phosphatidylethanolamine modified with polyethylene glycol or methoxypolyethylene glycol.

[0115] Among all the lipids constituting the liposome, the ratio of the PEG-modified lipid is preferably 1 to 15 mol%, and more preferably 2 to 10 mol%.

[0116] The liposome of the present invention preferably contains cholesterol. In the liposome, the addition of cholesterol is expected to reduce the fluidity of the liposome membrane by filling the gaps in the liposome membrane and the like.

[0117] The ratio of cholesterol in the lipids constituting the liposome is preferably 20 mol% to 50 mol%, more preferably 30 mol% to 45 mol%, and further preferably 35 to 43 mol%.

[0118] (Topotecan or its salt)

[0119] The liposome of the present invention contains topotecan or its salt. The chemical name of topotecan is (10-[(dimethylamino)methyl]-4-ethyl-4,9-dihydroxy-1H-pyrano[3',4':6,7]indolo[1,2-b]quinoline-3,14(4H,12H)dione, which is an anticancer agent having an inhibitory effect on topoisomerase activity. In the present invention, topotecan may be topotecan itself, a pharmaceutically acceptable salt, or a prodrug that releases topotecan in vivo. In the present invention, topotecan hydrochloride is preferably used.

[0120] The concentration of topotecan or its salt in the liposome can be determined, for example, by liquid chromatography / ultraviolet-visible absorbance detection method. And the concentration of sulfate ions in the inner aqueous phase of the liposome can be determined, for example, by ion chromatography.

[0121] The content of topotecan or its salt in the liposome composition is not particularly limited, and is preferably 0.025 to 20 mg / mL, more preferably 0.25 to 10 mg / mL, relative to the liposome composition.

[0122] From the viewpoints of the release rate of the liposome, the osmotic pressure inside the liposome, or the shape of the liposome caused by the precipitated drug, the molar ratio of topotecan or its salt contained in the liposome is preferably 0.1 to 1.5, more preferably 0.2 to 0.3, based on the amount of the lipid forming the liposome membrane.

[0123] When the molar ratio of topotecan or its salt to the lipid is too low, the area of the liposome membrane per unit drug amount becomes large, and thus the release rate of the drug from the liposome becomes fast, impairing the function of improving blood retention. On the other hand, when the molar ratio of topotecan or its salt to the lipid is too high, the osmotic pressure inside the liposome rises due to the increase in the dissolved amount of the drug, and thus the liposome is destroyed, or when the drug precipitates inside the liposome, the precipitated solid substance grows in large amounts, deforming the liposome shape.

[0124] (Inner aqueous phase)

[0125] The liposome composition of the present invention contains liposomes having an inner aqueous phase and an aqueous solution as an outer aqueous phase for dispersing the liposomes, and the liposome contains topotecan or its salt. An ammonium salt is preferably contained in the inner aqueous phase of the liposome. Examples of the ammonium salt include ammonium sulfate salt, ammonium citrate, ammonium phosphate, ammonium tartrate, ammonium succinate, fatty acid ammonium, ammonium chloride, sucrose 8 ammonium sulfate, etc. Among them, ammonium sulfate salt, ammonium citrate salt, ammonium phosphate salt, and sucrose 8 ammonium sulfate salt are preferred because they can improve the retention stability of the drug by precipitating it by reducing the solubility of the drug in the liposome, and ammonium sulfate salt is particularly preferred.

[0126] When ammonium sulfate is contained in the inner aqueous phase of the present invention, the molar ratio of sulfate ions in the inner aqueous phase to the total molar amount of topotecan as topotecan or its salt contained in the liposome composition of the present invention is preferably 0.36 or more, more preferably 0.4 or more, further preferably 0.4 or more and 1.8 or less, and particularly preferably 0.6 or more and 1.8 or less. By setting the molar ratio of sulfate ions as described above, leakage of topotecan or its salt from the liposome in the blood can be suppressed.

[0127] Further, when ammonium sulfate is contained in the inner aqueous phase of the present invention, the ratio of sulfate ions contained in ammonium sulfate to the total sulfate ions of the anti-tumor agent (inner aqueous phase ratio of sulfate ions) is preferably at least 80%, more preferably 90% or more. At the same time, the ratio of topotecan or its salt contained in the inner aqueous phase of the liposome to the total topotecan or its salt of the anti-tumor agent (inner aqueous phase ratio of the drug) is preferably at least 80%, more preferably 90% or more.

[0128] (Outer aqueous phase)

[0129] The liposome composition of the present invention contains liposomes having an inner aqueous phase and an aqueous solution as the outer aqueous phase for dispersing the liposomes, and the liposome contains topotecan or its salt. The pH of the outer aqueous phase of the liposome composition is preferably neutral, specifically, preferably about pH 5.5 to 8.5.

[0130] (Method for producing liposome composition)

[0131] The liposome of the present invention is not particularly limited. For example, it can be implemented by referring to Patent Document 1 (International Publication No. 2018 / 181963).

[0132] (Liposome composition)

[0133] The liposome composition of the present invention can contain at least one of a pharmaceutically acceptable isotonic agent, a stabilizer, an antioxidant, and a pH adjuster with respect to the administration route. That is, the liposome composition of the present invention can be provided as a pharmaceutical composition.

[0134] The isotonic agent is not particularly limited. For example, it can include inorganic salts such as sodium chloride, potassium chloride, sodium hydrogen phosphate, sodium dihydrogen phosphate, and potassium dihydrogen phosphate; polyhydric alcohols such as glycerol, mannitol, and sorbitol; and sugars such as glucose, fructose, lactose, or sucrose.

[0135] The stabilizer is not particularly limited. For example, it can include sugars such as glycerol, mannitol, sorbitol, lactose, or sucrose.

[0136] The antioxidant is not particularly limited. For example, it can include ascorbic acid, uric acid, tocopherol homologues (for example, vitamin E, and the four isomers of α, β, γ, and δ tocopherols), cysteine, EDTA (ethylenediamine tetraacetic acid), etc. The stabilizer and the antioxidant can be used alone or in combination of two or more.

[0137] Examples of the pH adjuster include sodium hydroxide, citric acid, acetic acid, triethanolamine, sodium hydrogen phosphate, sodium dihydrogen phosphate, and potassium dihydrogen phosphate.

[0138] The liposomal composition of the present invention may contain pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium carboxymethylcellulose, sodium polyacrylate, sodium alginate, water-soluble dextran, sodium carboxymethyl starch, pectin, methylcellulose, ethylcellulose, xanthan gum, gum arabic, casein, gelatin, agar, diglycerol, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin (HSA), mannitol, sorbitol, lactose, phosphate buffered saline (PBS), sodium chloride, saccharides, biodegradable polymers, serum-free media, and additives acceptable as pharmaceutical additives.

[0139] The container for filling the liposomal composition of the present invention is not particularly limited, and a material with low oxygen permeability is preferred. For example, plastic containers, glass containers, bags made of laminated films having a gas barrier layer such as aluminum foil, aluminum vapor deposition film, aluminum oxide vapor deposition film, silicon oxide vapor deposition film, polyvinyl alcohol, ethylene-vinyl alcohol copolymer, polyethylene terephthalate, polyethylene naphthalate, polyvinylidene chloride, etc. can be mentioned. According to need, it is also possible to shield light by using colored glass, bags using aluminum foil or aluminum vapor deposition film, etc.

[0140] In the container filled with the liposomal composition, in order to prevent oxidation due to oxygen present in the space part inside the container, it is preferred to displace the gas in the container space part and the liquid medicine with an inert gas such as nitrogen. For example, it can be mentioned that nitrogen is bubbled through the injection solution and it is filled into the container under a nitrogen atmosphere.

[0141] As the administration route of the liposomal composition of the present invention, parenteral administration is preferred. For example, intravenous injection (intravenous injection) such as drip, intramuscular injection, intraperitoneal injection, subcutaneous injection, intraocular injection, and intrathecal injection can be mentioned. As the administration method, administration by syringe or drip can be mentioned.

[0142] (Usage and dosage of the liposomal composition)

[0143] In the liposomal composition of the present invention, the dosage per administration of topotecan or its salt contained in the liposome, calculated as topotecan, is preferably 0.1 mg / m 2 body surface area to 10 mg / m 2 body surface area. More preferably, it is 0.5 mg / m 2 body surface area to 5 mg / m 2 body surface area, and further preferably 1.0 mg / m 2 body surface area to 3.5 mg / m 2 body surface area.

[0144] The liposomal composition of the present invention is preferably administered once every 1 to 8 weeks repeatedly. More preferably, it is administered once every 1 to 6 weeks repeatedly, further preferably once every 1 to 4 weeks repeatedly, and particularly preferably once every 2 weeks repeatedly.

[0145] When the liposomal composition of the present invention is administered once, it is preferably administered by infusion over 5 to 360 minutes. More preferably, it is 5 to 240 minutes, further preferably 10 to 120 minutes, and particularly preferably 30 to 120 minutes.

[0146] Regarding the dosage of topotecan or its salt contained in the liposomal composition of the present invention per administration, for example, in terms of topotecan, it is about 0.1 mg / m 2 body surface area, about 0.5 mg / m 2 body surface area, about 1.0 mg / m 2 body surface area, about 1.5 mg / m 2 body surface area, about 2.0 mg / m 2 body surface area, about 2.5 mg / m 2 body surface area, about 2.6 mg / m 2 body surface area, about 3.0 mg / m 2 body surface area, about 3.5 mg / m 2 body surface area, about 4.0 mg / m 2 body surface area, about 4.5 mg / m 2 body surface area, about 5.0 mg / m 2 body surface area, about 5.5 mg / m 2 body surface area, about 6.0 mg / m 2 body surface area, about 6.5 mg / m 2 body surface area, about 7.0 mg / m 2 body surface area, about 7.5 mg / m 2 body surface area, about 8.0 mg / m 2 body surface area, about 8.5 mg / m 2 body surface area, about 9.0 mg / m 2 body surface area, about 9.5 mg / m 2 body surface area, about 10 mg / m 2 body surface area. It is preferably about 0.5 mg / m 2 、about 1.0 mg / m 2 body surface area, about 1.5 mg / m 2 body surface area, about 2.0 mg / m 2 body surface area, about 2.5 mg / m 2 body surface area, about 2.6 mg / m2 body surface area, about 3.0 mg / m 2 body surface area, about 3.5 mg / m 2 body surface area, about 5.0 mg / m 2 body surface area, more preferably about 1.0 mg / m 2 body surface area, about 1.5 mg / m 2 body surface area, about 2.0 mg / m 2 body surface area, about 2.5 mg / m 2 body surface area, about 2.6 mg / m 2 body surface area, about 3.0 mg / m 2 body surface area, about 3.5 mg / m 2 body surface area, particularly preferably about 1.0 mg / m 2 body surface area, about 1.5 mg / m 2 body surface area, about 2.0 mg / m 2 body surface area, about 2.5 mg / m 2 body surface area, about 2.6 mg / m 2 body surface area, about 3.5 mg / m 2 body surface area.

[0147] (DNA damage repair inhibitor)

[0148] The drug of the present invention uses a DNA damage repair inhibitor (DNA damage response inhibitor) as a drug administered simultaneously or sequentially in combination with a liposomal composition containing topotecan or a salt thereof. The DNA damage repair inhibitor is a type of anticancer agent. Cancer cells rapidly proliferate by actively undergoing cell division. During proliferation, a large amount of gene (DNA) damage occurs, but in cancer, sometimes biomolecules that repair this damage strongly function. An anticancer agent that causes cancer cells to die by inhibiting the function of repairing their DNA damage is a DNA damage repair inhibitor.

[0149] Examples of the DNA damage repair inhibitor include PARP inhibitors, ATR inhibitors, ATM inhibitors, DNA-PK inhibitors, CHK1 / 2 inhibitors, and WEE1 inhibitors, and inhibitors that inhibit the pathways related to them, and these have the function of inhibiting biomolecules that repair DNA damage in cancer. Among the DNA damage repair inhibitors, in the drug of the present invention, PARP inhibitors, ATR inhibitors, ATM inhibitors, CHK1 / 2 inhibitors, WEE1 inhibitors, and DNA-PK inhibitors are preferred, ATR inhibitors, ATM inhibitors, and PARP inhibitors are more preferred, and PARP inhibitors are further preferred.

[0150] As PARP inhibitors, Olaparib, Talazoparib, Veliparib, Niraparib, Iniparib, Rucaparib, etc. can be mentioned, and Olaparib is preferred.

[0151] As ATR inhibitors, Ceralasertrib, Berzosertib, Elimusertib, M1774, RP-3500, ATRN-119, ART0380, IMP9064, HRS2398, M4344, BAY-1895344, etc. can be mentioned, and Ceralasertrib is preferred.

[0152] As ATM inhibitors, AZD0156, KU60019, AZD1390, M3541, M4076 can be mentioned, and M4076 is preferred.

[0153] As CHK1 / 2 inhibitors, Prexasertib, MK-8776, AZD7762, LY2603618, GDC-0575, SRA-737, ACR-368, etc. can be mentioned, and Prexasertib is preferred.

[0154] As DNA-PK inhibitors, M3814, CC-115, AZD7648, etc. can be mentioned, and M3814 is preferred.

[0155] As WEE1 inhibitors, ZN-c3, Adavosertib, Debio0123, IMP7068, SY4835, etc. can be mentioned, and ZN-c3 is preferred.

[0156] In the present invention, one or more DNA damage repair inhibitors can be used. The DNA damage repair inhibitors can be obtained by purchasing commercially available products.

[0157] The dosage and the number of administrations of the DNA damage repair inhibitor of the present invention can be appropriately set according to the type of the drug, the state of the patient, etc. For example, the mass of the drug per day as an active ingredient of the DNA damage repair inhibitor can be set in the range of about 0.1 mg to about 5000 mg. Further, the administration route of the DNA damage repair inhibitor can be oral or parenteral (for example, injection, drip, and administration to the rectal region, etc.), and the administration can be performed according to known clinical practices.

[0158] When the DNA damage repair inhibitor of the present invention is a PARP inhibitor, for example, the daily drug mass of the active ingredient as the PARP inhibitor can be set within the range of about 0.1 mg to about 5000 mg. When olaparib is used as the PARP inhibitor, it can be appropriately set according to the dosage and administration frequency described in the attachment of olaparib. For olaparib, it can be orally administered in the range of 30 to 500 mg each time.

[0159] When the DNA damage repair inhibitor of the present invention is an ATR inhibitor, for example, the daily drug mass of the active ingredient as the ATR inhibitor can be set within the range of about 0.1 mg to about 5000 mg. When ceralasertib is used as the ATR inhibitor, the dosage and administration frequency can be appropriately set according to the clinical performance.

[0160] When the DNA damage repair inhibitor of the present invention is an ATM inhibitor, for example, the daily drug mass of the active ingredient as the ATM inhibitor can be set within the range of about 0.1 mg to about 5000 mg. When M4076 is used as the ATM inhibitor, the dosage and administration frequency can be appropriately set according to the clinical performance.

[0161] When the DNA damage repair inhibitor of the present invention is a CHK1 / 2 inhibitor, for example, the daily drug mass of the active ingredient as the CHK1 / 2 inhibitor can be set within the range of about 0.1 mg to about 5000 mg. When prexasertib is used as the CHK1 / 2 inhibitor, the dosage and administration frequency can be appropriately set according to the clinical performance.

[0162] When the DNA damage repair inhibitor of the present invention is a DNA-PK inhibitor, for example, the daily drug mass of the active ingredient as the DNA-PK inhibitor can be set within the range of about 0.1 mg to about 5000 mg. When M3814 is used as the DNA-PK inhibitor, the dosage and administration frequency can be appropriately set according to the clinical performance.

[0163] When the DNA damage repair inhibitor of the present invention is a WEE1 inhibitor, for example, the daily drug mass of the active ingredient as the WEE1 inhibitor can be set within the range of about 0.1 mg to about 5000 mg. When ZN-c3 is used as the WEE1 inhibitor, the dosage and administration frequency can be appropriately set according to the clinical performance.

[0164] The drug of the present invention is a drug that combines a liposome composition containing topotecan or its salt and a DNA damage repair inhibitor and administers them simultaneously or sequentially, and is preferably used as an anticancer agent.

[0165] The cancer that is the target of the drug of the present invention is preferably solid carcinoma. As solid carcinoma, ovarian cancer, uterine cancer, lung cancer, Merkel cell carcinoma, skin cancer, breast cancer, malignant soft tumor, neuroendocrine tumor, brain tumor, pharyngeal cancer, laryngeal cancer, thyroid cancer, esophageal cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, gallbladder cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone tumor can be mentioned, and preferably at least one selected from breast cancer, uterine cancer, ovarian cancer, lung cancer, Merkel cell carcinoma, neuroendocrine tumor, brain tumor.

[0166] As ovarian cancer, serous ovarian cancer, endometrioid ovarian cancer, clear cell ovarian cancer, mucinous ovarian cancer can be mentioned, and serous ovarian cancer and endometrioid ovarian cancer are particularly preferred. Moreover, as ovarian cancer, it can be ovarian cancer resistant to platinum preparations.

[0167] As uterine cancer, cervical cancer, uterine cancer, uterine sarcoma can be mentioned, and cervical cancer and uterine sarcoma are particularly preferred.

[0168] As lung cancer, non-small cell lung cancer, small cell lung cancer can be mentioned, and small cell lung cancer is particularly preferred.

[0169] It can be seen that by combining the liposome composition containing topotecan or its salt and the DNA damage repair inhibitor and administering them simultaneously or sequentially, the drug of the present invention has a stronger anti-tumor effect (for example, tumor growth inhibitory effect, etc.) compared with each single drug (the liposome composition containing topotecan or its salt or the DNA damage repair inhibitor).

[0170] The mechanism of action of the combined use of the liposome composition of the present invention and DNA damage repair inhibition is speculated as follows, but is not limited to the following.

[0171] The topotecan or its salt contained in the liposome composition of the present invention is a drug known as a topoisomerase I inhibitor. During cell division, topoisomerase I has the function of cleaving / recombining one of the double helix structures of DNA. By inhibiting this topoisomerase I, DNA damage can be actively caused.

[0172] Moreover, by incorporating topotecan or its salt into liposomes, damage to the bone marrow can be reduced while the damage effect on DNA in cancer cells is not weakened.

[0173] The drug of the present invention can also administer the DNA damage repair inhibitor after administering the liposome composition containing topotecan or its salt. By administering the DNA damage repair inhibitor after administering the liposome composition of the present invention, the therapeutic effect can be maintained, and the recurrence of cancer and the progression of cancer can be prevented, thereby improving the QOL of patients can be expected.

[0174] The drug of the present invention can also administer a liposome composition containing topotecan or a salt thereof after administering a DNA damage repair inhibitor. By administering a DNA damage repair inhibitor after administering the liposome composition of the present invention, the therapeutic effect can be maintained, and the recurrence of cancer can be prevented and the progression of cancer can be prevented, so that an improvement in the QOL of patients can be expected.

[0175] The ratio of the dosage or blending amount of the liposome composition containing topotecan or a salt thereof and the DNA damage repair inhibitor in the drug of the present invention is not particularly limited as long as it is within the range of exerting an enhancing effect on the therapeutic effect on cancer.

[0176] For example, the dosage of topotecan or a salt thereof in the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of the DNA damage repair inhibitor, preferably set to about 0.000001 to 10 times (mass ratio) of the DNA damage repair inhibitor, more preferably set to about 0.00001 to 1 times (mass ratio) of the DNA damage repair inhibitor, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of the DNA damage repair inhibitor.

[0177] When using a PARP inhibitor as the DNA damage repair inhibitor, the dosage of the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of the PARP inhibitor, preferably set to about 0.000001 to 10 times (mass ratio) of the PARP inhibitor, more preferably set to about 0.00001 to 1 times (mass ratio) of the PARP inhibitor, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of the PARP inhibitor.

[0178] Among them, when using olaparib as the PARP inhibitor, the dosage of the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of olaparib, preferably set to about 0.000001 to 10 times (mass ratio) of olaparib, more preferably set to about 0.00001 to 1 times (mass ratio) of olaparib, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of olaparib.

[0179] When using an ATR inhibitor as the DNA damage repair inhibitor, the dosage of the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of the ATR inhibitor, preferably set to about 0.000001 to 10 times (mass ratio) of the ATR inhibitor, more preferably set to about 0.00001 to 1 times (mass ratio) of the ATR inhibitor, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of the ATR inhibitor.

[0180] When using an ATM inhibitor as a DNA damage repair inhibitor, the dosage of the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of the ATM inhibitor, preferably set to about 0.000001 to 10 times (mass ratio) of the ATM inhibitor, more preferably set to about 0.00001 to 1 time (mass ratio) of the ATM inhibitor, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of the ATM inhibitor.

[0181] When using a CHK1 / 2 inhibitor as a DNA damage repair inhibitor, the dosage of the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of the CHK1 / 2 inhibitor, preferably set to about 0.000001 to 10 times (mass ratio) of the CHK1 / 2 inhibitor, more preferably set to about 0.00001 to 1 time (mass ratio) of the CHK1 / 2 inhibitor, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of the CHK1 / 2 inhibitor.

[0182] When using a DNA-PK inhibitor as a DNA damage repair inhibitor, the dosage of the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of the DNA-PK inhibitor, preferably set to about 0.000001 to 10 times (mass ratio) of the DNA-PK inhibitor, more preferably set to about 0.00001 to 1 time (mass ratio) of the DNA-PK inhibitor, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of the DNA-PK inhibitor.

[0183] When using a WEE1 inhibitor as a DNA damage repair inhibitor, the dosage of the liposome composition of the present invention is set to about 0.0000001 to 100 times (mass ratio) of the WEE1 inhibitor, preferably set to about 0.000001 to 10 times (mass ratio) of the WEE1 inhibitor, more preferably set to about 0.00001 to 1 time (mass ratio) of the WEE1 inhibitor, and further preferably set to about 0.0001 to 0.1 times (mass ratio) of the WEE1 inhibitor.

[0184] The present invention will be further described in detail by the following examples, but the present invention is not limited to these examples.

[0185] Examples

[0186] Fully synthetic DHSM refers to dihydrosphingomyelin produced by chemical synthesis, and the dihydrosphingomyelin contains more than 98% of the following compounds having long-chain alkyl groups with 16 and 18 carbon atoms.

[0187] [Chemical Formula 2]

[0188]

[0189] As the PEG phospholipid (labeled as PEG in the table), SUNBRIGHT DSPE-020CN manufactured by NOF CORPORATION (hereinafter referred to as DSPE-PEG) was used.

[0190] As cholesterol (labeled as Chol in the table), Cholesterol HP (manufactured by NIPPON FINE CHEMICAL CO., LTD.) was used.

[0191] <Reference Example 1> Preparation of Liposome Composition Containing Topotecan or Its Salt

[0192] (a) Preparation of Oil Phase

[0193] Regarding Example 1, 28.63 g, 9.94 g, and 9.94 g of fully synthetic DHSM, PEG phospholipid (SUNBRIGHT DSPE-020CN, manufactured by NOF CORPORATION, hereinafter referred to as DSPE-PEG), and cholesterol were weighed, respectively. The lipids were mixed with 711 mL of ethanol and dissolved at 68 °C to prepare an oil phase.

[0194] (b1) Preparation of Aqueous Phase 1

[0195] 58.0 g of ammonium sulfate was dissolved in 2573.0 g of water to prepare aqueous phase 1.

[0196] (b2) Preparation of Aqueous Phase 2

[0197] 11.6 g of ammonium sulfate was dissolved in 514.5 g of water to prepare aqueous phase 2.

[0198] (c) Formation of Liposome Particles by Emulsification

[0199] The aqueous phase 1 prepared in (b1) was heated to 68 °C, and after adding all of the oil phase prepared in (a), it was mixed for 60 minutes at a circumferential speed of 26 m / s using a precision emulsifying disperser. Then, the liquid temperature was adjusted to 56 °C and mixed for 30 minutes. Then, after adding the aqueous phase 2 at room temperature, while heating at 65 °C, stirring was continued slowly by rotary immersion, thereby evaporating the organic solvent and water, and heating and stirring were stopped when the osmotic pressure of the liquid was concentrated to 0.8 mS / cm to stop the evaporation.

[0200] (e) Replacement of the Outer Aqueous Phase Liquid of Liposomes by Dialysis

[0201] As the dialysate, an aqueous solution of 3.15% by mass of NaCl was used. Using this dialysate, the liquid obtained in (c) was subjected to cross flow filtration at room temperature to remove ammonium sulfate present in the outer aqueous phase, and liposomes with the outer aqueous phase replaced by the dialysate were obtained.

[0202] (f) Incorporation of topotecan into liposome particles by remote loading

[0203] Water for injection was added to topotecan hydrochloride (manufactured by ScinoPharm Taiwan Ltd) to make it 5.9 mg / mL. Further, while stirring the liquid well, 1 mol / L NaOH solution was added to adjust the pH to about 3 and dissolve topotecan. After adding liposomes to this topotecan solution at a volume ratio of 1 / 1, it was heated at 62 °C for 30 minutes.

[0204] (g) Removal of topotecan from the outer aqueous phase by dialysis

[0205] As the dialysate, a sucrose / histidine buffer solution composed of 9.4% by mass of sucrose and 10 mmol / L of histidine was prepared. Using this dialysate, the solution obtained in (f) was subjected to cross flow filtration at room temperature to remove topotecan present in the outer aqueous phase, and liposomes containing topotecan with the outer aqueous phase replaced by the dialysate were obtained.

[0206] [Physical property measurement and evaluation]

[0207] <Average particle size>

[0208] In the present invention, the average particle size refers to the cumulative average particle size measured by dynamic light scattering method. The cumulative average particle size measured by dynamic light scattering method using ELSZ-2000ZS (manufactured by Otsuka Electronics Co., Ltd.) was 114 nm.

[0209] (Topotecan concentration measurement results)

[0210] The result of measuring the sample using an HPLC (high performance liquid chromatography) device 1290DAD (manufactured by Agilent) and quantifying the topotecan concentration was 104.8%. And, relative to the amount of topotecan contained in the total liposomes, the topotecan contained inside the liposomes was 99.9%. The detailed content of the measurement method is described below.

[0211] (Measurement of all topotecan amounts in the liposome preparation)

[0212] The sample solution was prepared by dissolving the prepared liposome liquid in methanol containing 0.1% trifluoroacetic acid, and the calibration curve standard solution was prepared by diluting topotecan hydrochloride. The determination was carried out by liquid chromatography / ultraviolet-visible absorbance detection method.

[0213] The concentration of topotecan in the inner aqueous phase was calculated by subtracting the concentration of topotecan in the outer aqueous phase from the total aqueous phase topotecan concentration.

[0214] (Adjustment of the sample for determining the concentration of topotecan in the outer aqueous phase)

[0215] 100 μL of the liposome dispersion was measured and diluted by adding 900 μL of 1×PBS. 500 μL of this dispersion was weighed and treated with an ultrafiltration filter (7400×g, 5°C, 60 minutes), and the filtrate was used as the HPLC analysis sample. The centrifuge used was the Universal cooling centrifuge model 5922 manufactured by KUBOTA Corporation.

[0216] (Preparation of the calibration curve standard solution)

[0217] Approximately 15 mg of topotecan hydrochloride was weighed, and 100 mL of methanol containing 0.1% trifluoroacetic acid was added to obtain the stock solution of the standard solution. 2 mL of this stock solution was weighed and 50 mL of methanol containing 0.1% trifluoroacetic acid was added to obtain the calibration curve standard solution.

[0218] (Determination)

[0219] The determination was carried out by liquid chromatography / ultraviolet-visible absorbance detection method under the following conditions.

[0220] Detection wavelength: 382 nm, column: ACQUITY UPLC CSH Fluoro-Phenyl, particle size 1.7 μM, inner diameter 2.1 mm, length 10 cm (Waters (water column))

[0221] Column temperature: Constant temperature of about 40°C

[0222] The mobile phase was a mixture of water / methanol / trifluoroacetic acid, and the concentration gradient of the mobile phase was controlled by changing the mixing ratio of each solvent during the liquid delivery of the mobile phase.

[0223] The determination was carried out at a flow rate of 1.0 mL per minute, an injection volume of 12 μL, and an autosampler temperature of constant temperature of about 10°C.

[0224] <Determination of sulfate ion concentration>

[0225] The sample was measured using an ion chromatograph 883 Basic IC plus (manufactured by Metrohm), and the concentration of sulfate ions was quantified. The sulfate ions contained therein were shown to be 0.065 w / v%. It was found that the sulfate ions present in the external aqueous phase were 0.0003 w / v% and most of them were contained within. The molar ratio of the amount of sulfate ions contained within to the amount of topotecan contained in the total aqueous phase was 1.19.

[0226] The concentration of sulfate ions in the internal aqueous phase was calculated by subtracting the concentration of sulfate ions in the external aqueous phase from the concentration of sulfate ions in the total aqueous phase. The concentration of sulfate ions in each aqueous phase was measured as follows.

[0227] (Concentration of sulfate ions in the total aqueous phase)

[0228] 100 μL of the liposome dispersion was taken, and methanol was added to make it exactly 10 mL and mixed. 1000 μL of this liquid was taken and diluted to exactly 20 mL with a dilution solvent composed of 3% glycerol, 12 mmol / L sodium bicarbonate, and 0.6 mmol / L sodium carbonate. This solution was passed through a solid phase extraction cartridge oasis hlb plus light cartridge, each cartridge containing 30 mg of adsorbent (water column), and the fraction was used for ion chromatographic analysis.

[0229] (Concentration of sulfate ions in the external aqueous phase)

[0230] 100 μL of the liposome dispersion was taken and mixed with 900 μL of the above dilution solvent. 500 μL of this liquid was treated with an ultrafiltration filter AmiconUltra-0.5 (molecular weight cut-off 10k) (Merck Millipore), and the filtrate was used as a sample for ion chromatographic analysis.

[0231] The centrifugation conditions were 7400 g, 5 °C, and 30 minutes. A Universal cooling centrifuge model 5922 manufactured by KUBOTA Corporation was used.

[0232] The liposome composition of the present invention was manufactured according to the above-described examples. It was confirmed that the molar ratio of sulfate ions in the internal aqueous phase to the drug in the total aqueous phase was 0.36 or more.

[0233] [Test Example 1]

[0234] Pharmacodynamic and safety test when using the liposome composition of the present invention and olaparib as a PARP inhibitor in an established pancreatic cancer cell line (Capan-1) model

[0235] As the test substance, the liposome composition containing topotecan prepared in Reference Example 1 (hereinafter, also referred to as Lipo) was used. For the dilution of Lipo, a 5% glucose injection solution (manufactured by Otsuka Pharmaceutical Co., Ltd.) (hereinafter, also referred to as Lipo diluent) was used.

[0236] <Example 1>

[0237] 1×10 7 Capan-1 cells, which are an established human pancreatic cancer cell line, were transplanted subcutaneously into the right flank of female Balb / cAJcl-nu / nu mice to form subcutaneous tumors. Intravenous administration (i.v.) of 0.5 mg / kg of Lipo diluted to 0.05 mg / mL with a 5% glucose injection solution was performed on them. This administration was repeated once a week for 4 weeks. The first administration day was set as Day 0. In addition, intraperitoneal administration (i.p.) of 50 mg / kg of olaparib (MedChem Express) diluted with PBS containing 10% DMSO in 10% HP-βCYD was performed to reach 5 mg / mL on Day 0. This administration was repeated once a day for 6 times, and after a 1-day drug withdrawal, the same administration was continued and repeated for a total of 4 weeks. Using the tumor volume as an index, the inhibitory effect of the tumor by the administration of the present invention was evaluated. And, as an index of safety, the body weight change was measured.

[0238] <Comparative Example 1>

[0239] Except for administering only the same volume of the dilution solvent without administering Lipo and olaparib, it was carried out in the same manner as in Example 1. That is, no drug was administered.

[0240] <Comparative Example 2>

[0241] Except for administering only the same volume of the dilution solvent without administering Lipo, it was carried out in the same manner as in Example 1. That is, the only drug administered was olaparib.

[0242] <Comparative Example 3>

[0243] Except for administering only the same volume of the dilution solvent without administering olaparib, it was carried out in the same manner as in Example 1. That is, the only drug administered was Lipo.

[0244] <Comparative Example 4>

[0245] Lipo was not administered, and intravenously administered (i.v.) 5 mg / kg of topotecan (manufactured by ScinoPharm Taiwan Ltd) diluted to 0.5 mg / mL with physiological saline (manufactured by Otsuka Pharmaceutical Co., Ltd.) was performed. This administration was repeated once a week for 4 weeks. Also, olaparib was not administered and only the same volume of the dilution solvent was administered. Except for the above, it was carried out in the same manner as in Example 1. That is, the administered agent was topotecan not contained in liposomes.

[0246] <Comparative Example 5>

[0247] Lipo was not administered, and intravenously administered (i.v.) 5 mg / kg of topotecan (manufactured by ScinoPharm Taiwan Ltd) diluted to 0.5 mg / mL with physiological saline (manufactured by Otsuka Pharmaceutical Co., Ltd.) was performed. This administration was repeated once a week for 4 weeks. Except for this, it was carried out in the same manner as in Example 1. That is, the administered agents were topotecan not contained in liposomes and olaparib.

[0248] <Administration Conditions of Example 1 and Comparative Examples 1 to 5>

[0249] The administration conditions of Example 1 and Comparative Examples 1 to 5 were summarized in Table 1.

[0250] [Table 1]

[0251]

[0252] <Evaluation Results of Efficacy and Safety of Example 1 and Comparative Examples 1 to 5>

[0253] In Table 2, Figure 1 and Figure 2 show the evaluation results of the efficacy (tumor diameter) and safety (body weight) of Example 1 and Comparative Examples 1 to 5.

[0254] [Table 2]

[0255]

[0256] Compared with Comparative Example 1 in which no agent was administered after 28 days of administration, in Example 1 of the present invention, the tumor diameter was 7%, showing a very strong anti-tumor effect. On the other hand, it was found that there was almost no difference in body weight, and this administration also showed excellent safety.

[0257] Moreover, even when compared with Comparative Example 3 that administered only Lipo or Comparative Example 5 that combined topotecan not contained in liposomes with olaparib, the antitumor effect of Example 1 was high. Furthermore, from the viewpoint of safety with body weight change as an index, compared with Comparative Example 1 that did not administer the agent, a significant 15% reduction in body weight was observed in Comparative Example 5 that combined and administered topotecan and olaparib. It is considered that the reason is that strong toxicity was produced due to the combination of topotecan and olaparib. On the other hand, in the combination of liposomal topotecan (Lipo) and olaparib, such enhanced toxicity was not observed. Thus, the combination of the liposomal topotecan of the present invention and a DNA damage repair inhibitor such as olaparib is an amazing invention that can prevent enhanced toxicity and improve drug efficacy.

[0258] [Test Example 2]

[0259] Pharmacodynamic and safety test when olaparib, a PARP inhibitor, was successively administered after the administration of the liposomal composition of the present invention in an established pancreatic cancer cell line (Capan-1) model

[0260] As the test substance, the liposomal composition prepared in Reference Example 1 (hereinafter, also referred to as Lipo) was used in the same manner as in Example 1. A 5% glucose injection solution (manufactured by Otsuka Pharmaceutical Co., Ltd.) (hereinafter, also referred to as Lipo diluent) was used for the dilution of Lipo.

[0261] <Example 2>

[0262] 3×10 6 Capan-1 cells, which are an established human pancreatic cancer cell line, were transplanted subcutaneously into the right abdominal flank of female Balb / cAJcl-nu / nu mice to form subcutaneous tumors. Intravenous administration (i.v.) of 2 mg / kg of Lipo diluted to 0.05 mg / mL with a 5% glucose injection solution was performed on them. This administration was repeated once a week for 4 weeks. That is, if the first administration day was set as day 0, administration was performed on day 0, day 7, day 14, and day 21. In addition, intraperitoneal administration (i.p.) of 50 mg / kg of olaparib (MedChem Express) diluted with PBS containing 10% DMSO in 10% HP-βCYD was performed to reach 5 mg / mL on day 24, 3 days after the end of the administration of Lipo. This administration was repeated once a day for 5 times, and after a 1-day drug withdrawal, the same administration was continued for a total of 4 weeks. Using the tumor volume as an index, the inhibitory effect of the tumor by the administration of the present invention was evaluated. And, as an index of safety, the body weight change was measured.

[0263] <Example 3>

[0264] Intraperitoneal administration (i.p.) of 50 mg / kg of olaparib (MedChem Express) diluted with PBS containing 10% DMSO in 10% HP-βCYD was performed to achieve 5 mg / mL on Day 0 simultaneous with the administration of Lipo. This administration was repeated once a day for 5 times, and after a 1-day drug withdrawal, the same administration was continued for a total of 4 weeks. Except as described above, it was carried out in the same manner as in Example 2.

[0265] <Comparative Example 6>

[0266] Olaparib was not administered, and only the same volume of the dilution solvent was administered. Except for this, it was carried out in the same manner as in Example 2. That is, the only drug administered was Lipo.

[0267] <Administration Conditions of Examples 2 - 3 and Comparative Example 6>

[0268] The administration conditions of Examples 2 - 3 and Comparative Example 6 are tabulated in Table 3.

[0269] [Table 3]

[0270]

[0271] <Evaluation Results of Efficacy and Safety of Examples 2 - 3 and Comparative Example 6>

[0272] In Table 4, Figure 3 and Figure 4 show the evaluation results of the efficacy (tumor diameter) and safety (body weight) of Examples 2 - 3 and Comparative Example 6. The body weight shows the change when the first day of drug administration is set as 100%.

[0273] [Table 4]

[0274]

[0275] In Comparative Example 6 where only Lipo was administered, starting from 21 days after the end of Lipo administration, on the 99th day after a long period, the tumor grew significantly. On the other hand, in Examples 2 and 3 of the present invention, starting from 21 days after the end of Lipo administration, even on the 99th day after a long period, the tumor diameter was maintained small, and a strong tumor growth inhibitory effect was observed. It is considered that the reason is that the DNA damage repair inhibitor containing olaparib inhibits the repair of DNA damaged by topotecan and maintains a long-term tumor growth inhibitory effect.

[0276] In Example 3 where the administration periods of Lipo and olaparib overlapped, the tumor diameter on the 99th day was indeed smaller than that in Example 2 and showed a stronger anti-tumor effect. However, on the 25th day of administration, the body weight decreased relatively significantly. On the other hand, in Example 2, no significant decrease in body weight was observed, and relatively, the safety was higher than that in Example 3. Therefore, it can be understood that when safety is emphasized, as shown in Example 2, if olaparib is administered after the administration of Lipo is completed, it can be controlled.

[0277] [Test Example 3]

[0278] Evaluation of anti-tumor activity when the liposome composition of the present invention is used in combination with a DNA damage repair inhibitor

[0279] To evaluate the synergistic effect produced by combining the liposome composition of the present invention with various DNA damage repair inhibitors, as an in vitro cell proliferation inhibition test, the test was carried out using the cell types shown in Table 5, the number of cells inoculated in wells, and the culture conditions. As a method for detecting living cells, Cell Counting Kit-8 (DOJINDO LABORATORIES) was used, and

[0280] the absorbance at 460 nm was measured using a microplate reader 4 hours after adding the reagent, and the IC 50 value was thus obtained.

[0281] [Table 5]

[0282]

[0283] As various DNA damage repair inhibitors used in combination with the liposome composition, the ATR inhibitor Ceralasertib (Selleck Biotech), the CHK1 / 2 inhibitor Prexasertib (Selleck Biotech), the DNA-PK inhibitor M3814 (Selleck Biotech), the ATM inhibitor M4076 (Selleck Biotech), the WEE1 inhibitor ZN-c3 (Selleck Biotech), and the PARP inhibitor Olaparib (Selleck Biotech) were used. The added concentrations of Lipo and various DNA damage repair inhibitors prepared in Reference Example 1 for each cell type are shown in Tables 6 and 7.

[0284] [Table 6]

[0285]

[0286] [Table 7]

[0287]

[0288] Regarding the synergistic effect generated by combination, it was evaluated by the isobologram analysis commonly used for judging the combination effect. First, the IC 50 values of Lipo alone and various DNA damage repair inhibitors alone were determined. Next, various DNA damage repair inhibitors were combined at each concentration point of Lipo, and the IC 50 values of the DNA damage repair inhibitors were determined. The value obtained by dividing this by the IC 50 value of Lipo alone was set as "Lipo FIC", and the value obtained by dividing this by the IC 50 value of various DNA damage repair inhibitors alone was set as "various DNA damage repair inhibitor FIC", and these two values were recorded as a set of values. As a part of this, the respective FICs when combining Lipo and the ATR inhibitor Ceralasertib in Capan-1 are shown in Table 8.

[0289] Furthermore, Lipo was combined at each concentration point of various DNA damage repair inhibitors set in the same manner, and the IC 50 value of this Lipo was determined. The value obtained by dividing this by the IC 50 value of Lipo alone was set as "Lipo FIC", and the value obtained by dividing this by the IC 50 value of various DNA damage repair inhibitors alone was set as "various DNA damage repair inhibitor FIC". As a part of recording these two values as a set of values, the respective FICs when combining Lipo and the ATR inhibitor Ceralasertib in Capan-1 are shown in Table 9.

[0290] [Table 8]

[0291]

[0292] [Table 9]

[0293]

[0294] As shown in Table 8 and Table 9, these sets of values were respectively plotted on a graph of various DNA damage repair inhibitor FIC and Lipo FIC, thereby obtaining an interaction effect diagram. In the interaction effect diagram, when plotted on the left side compared to the straight line connecting the points where the FICs of each axis are 1, it shows a "synergistic effect" and is regarded as a preferred combination effect. When on the straight line, it shows an "additive effect". When plotted on the right side, it shows an "antagonistic effect" and is regarded as an inappropriate combination effect. In Figures 5 to 21The results are shown therein.

[0295] [Research results of Capan-1]

[0296] In Figure 5 the test results of Ceralasertib are shown, in Figure 6 the test results of M4076 are shown, in Figure 7 the test results of M3814 are shown, in Figure 8 the test results of ZN-c3 are shown, and in Figure 9 the test results of Olaparib are shown. As Figure 5 , Figure 7 and Figure 9 shown, a clear synergistic effect was observed in Ceralasertib, M3814, and Olaparib. Also, as Figure 6 and Figure 8 shown, an additive effect was observed in M4076 and ZN-c3.

[0297] [Research results of ES-2]

[0298] In Figure 10 the test results of Ceralasertib are shown, in Figure 11 the test results of M4076 are shown, in Figure 12 the test results of Preaxasertib are shown, in Figure 13 the test results of M3814 are shown, in Figure 14 the test results of ZN-c3 are shown, and in Figure 15 the test results of Olaparib are shown. As Figure 10 , Figure 11 , Figure 12 , Figure 14 and Figure 15 shown, a clear synergistic effect was observed in Ceralasertib, Prexasertib, M4076, ZN-c3, and Olaparib. Also, as Figure 13 shown, an additive effect was observed in M3814.

[0299] [Research results of DMS114]

[0300] In Figure 16 the test results of Ceralasertib are shown, in Figure 17 the test results of M4076 are shown, in Figure 18 the test results of Prexasertib are shown, in Figure 19 the test results of M3814 are shown, in Figure 20The test results of ZN-c3 are shown, as well as those of Olaparib shown in Figure 21 As shown in Figure 16 , Figure 17 , Figure 18 , Figure 19 and Figure 21 shown, a clear synergistic effect was observed in Ceralasertib, Prexasertib, M3814, M4076, and Olaparib. Also, as shown in Figure 20 , an additive effect was observed in ZN-c3.

[0301] [Test Example 4]

[0302] Test on the combined effect of the liposome composition of the present invention and a DNA damage repair inhibitor in cancer patients

[0303] As the test substance, referring to Patent Document 2 (International Publication No. 2018 / 181963), a liquid pharmaceutical preparation containing liposomes containing topotecan with the following composition was prepared (hereinafter, referred to as Preparation A).

[0304] Topotecan hydrochloride 3.0 mg / mL (6.5 mmol)

[0305] Fully synthetic DHSM 10.4 mg / mL

[0306] DSPE-MPEG2000 3.6 mg / mL

[0307] Cholesterol 3.6 mg / mL

[0308] Ammonium sulfate 0.9 mg / mL (6.9 mmol)

[0309] Sucrose appropriate amount

[0310] L-Histidine appropriate amount

[0311] Sodium chloride appropriate amount

[0312] Water (solvent) appropriate amount

[0313] <Physical property values of Preparation A>

[0314] The pH of Preparation A is 7.4, and the particle size of the liposomes contained in Preparation A is 91 nm in terms of the cumulative average particle size. Moreover, the molar ratio of sulfate ions contained in the inner aqueous phase to topotecan hydrochloride is 1.05.

[0315] <Determination of administration and therapeutic effect>

[0316] For cancer patients, Preparation A is administered once every 1 to 8 weeks so that the single-dose amount of topotecan reaches 0.1 mg / m 2 body surface area to 10 mg / m 2 body surface area. And, under the guidance of a doctor, a DNA damage repair inhibitor is administered according to known clinical practice. For example, when the DNA damage repair inhibitor is olaparib, it is orally administered in the range of 30 to 500 mg each time.

[0317] The therapeutic effect can be determined according to the following criteria.

[0318] The evaluation object is confirmed by image diagnosis based on MRI (magnetic resonance imaging), and the determination is made according to the following criteria.

[0319] CR (Complete Response): A state in which the tumor has completely disappeared

[0320] PR (Partial Response): A state in which the sum of the sizes of the tumors has decreased by more than 30%

[0321] SD (Stable Disease): A state in which the size of the tumor has not changed

[0322] PD (Progressive Disease): A state in which the sum of the sizes of the tumors has increased by more than 20% and the absolute value has also increased by more than 5 mm, or a state in which new lesions have appeared

[0323] The drug of the present invention can achieve excellent anti-tumor effects. Specifically, it does not increase side effects on normal organs, bone marrow, etc., and has a therapeutic effect on cancer (effects such as cancer volume reduction or no change in tumor size). And, effects such as preventing cancer recurrence and preventing cancer progression can also be expected by maintaining the therapeutic effect.

[0324] Based on these results, the drug of the present invention can be expected to extend the progression-free survival or overall survival of patients, and has a very useful effect from the viewpoint of improving the QOL of patients.

[0325] Industrial applicability

[0326] The drug of the present invention is useful for showing excellent therapeutic effects on cancer.

Claims

1. A drug, the combination of which contains (A) a liposome composition and (B) a DNA damage repair inhibitor, and the liposome composition and the DNA damage repair inhibitor are administered simultaneously or sequentially, wherein the (A) liposome composition contains liposomes having an inner aqueous phase and an aqueous solution as the outer aqueous phase for dispersing the liposomes, wherein the liposomes contain topotecan or a salt thereof, and the lipids constituting the liposomes contain dihydrosphingomyelin.

2. The drug according to claim 1, wherein, after administering the (A) liposome composition, the (B) DNA damage repair inhibitor is administered.

3. The drug according to claim 1, wherein, after administering the (B) DNA damage repair inhibitor, the (A) liposome composition is administered.

4. The drug according to any one of claims 1 to 3, wherein, the DNA damage repair inhibitor is at least one selected from PARP inhibitors, ATR inhibitors, ATM inhibitors, CHK1 / 2 inhibitors, WEE1 inhibitors, DNA-PK inhibitors, and inhibitors that inhibit the pathways related to them.

5. The drug according to any one of claims 1 to 3, wherein, the lipids constituting the liposomes further include cholesterol and lipids modified with polyethylene glycol.

6. The drug according to any one of claims 1 to 3, wherein, relative to the total amount of the lipids constituting the liposomes, the blending ratio of the lipids modified with polyethylene glycol is 2 mol% to 10 mol%.

7. The drug according to any one of claims 1 to 3, wherein, the lipids modified with polyethylene glycol are diacyl phosphatidylethanolamines modified with polyethylene glycol or methoxypolyethylene glycol.

8. The drug according to any one of claims 1 to 3, wherein, relative to the total amount of the lipids constituting the liposomes, the blending ratio of cholesterol is 35 to 43 mol%.

9. The drug according to any one of claims 1 to 3, wherein, the inner aqueous phase of the liposomes contains an ammonium salt.

10. The drug according to any one of claims 1 to 3, wherein, the DNA damage repair inhibitor is a PARP inhibitor.

11. The drug according to any one of claims 1 to 3, wherein, (A) The dosage of topotecan or its salt contained in the liposome composition is 0.1 mg / m 2 body surface area to 10 mg / m 2 body surface area.

12. The drug according to any one of claims 1 to 3, wherein, (A) The liposome composition is administered once every 1 to 8 weeks.

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