Pharmaceutical for enhancing therapeutic effect of immune checkpoint inhibitor re-administered to cancer patient who has discontinued administration of immune checkpoint inhibitor

Abstract

The present invention provides a pharmaceutical comprising, as an active ingredient, an agent that promotes the expression of Meflin. Said pharmaceutical is characterized by being used for a cancer patient who has discontinued administration of a first immune checkpoint inhibitor, and being administered in combination with a second immune checkpoint inhibitor, so as to enhance the therapeutic effect of the second immune checkpoint inhibitor. The present invention makes it possible to enhance the therapeutic effect of an immune checkpoint inhibitor re-administered to a cancer patient who has discontinued administration of the immune checkpoint inhibitor.

Description

A drug for improving the therapeutic effect of immune checkpoint inhibitors, to be reused in cancer patients who have discontinued the administration of immune checkpoint inhibitors

[0001] The present invention relates to a pharmaceutical for improving the therapeutic effect of an immune checkpoint inhibitor, which is reused in cancer patients who have discontinued administration of the immune checkpoint inhibitor.

[0002] Immune checkpoint inhibitors (ICIs) have been approved for first- or second-line or later treatment of many malignancies, either as monotherapy or in combination with anticancer drugs or molecularly targeted drugs, and their indications are expanding. However, if resistance develops after ICI therapy, ICIs are generally not used in subsequent lines of treatment, limiting options for such patients. Regarding the re-use of ICIs after ICI therapy (ICI rechallenge), studies of solid tumors such as non-small cell lung cancer have shown wide variation in opinion, and no conclusion has been reached regarding the efficacy of ICI rechallenge (NPLs 1-3). As a result, the National Comprehensive Cancer Network (NCCN) guidelines (NPL 4) currently do not recommend ICI rechallenge.

[0003] Akamatsu H et al. Nivolumab Retreatment in Non-Small Cell Lung Cancer Patients Who Responded to Prior Immune Checkpoint Inhibitors and Had ICI-Free Intervals (WJOG9616L). Clin Cancer Res. 2022 Jun 28;28(15):OF1-OF7.Plazy C, Hannani D, Gobbini E. Immune Checkpoint Inhibitor Rechallenge and Resumption: a Systematic Review. Curr Oncol Rep. 2022 Sep;24(9):1095-1106.Perdyan A et al. The Effectiveness of Cancer Immune Checkpoint Inhibitor Retreatment and Rechallenge-A Systematic Review. Cancers (Basel). 2023 Jul 4;15(13):3490.National Comprehensive Cancer Network Guidelines Version 5.2024, Non-Small Cell Lung Cancer

[0004] An objective of the present invention is to provide a pharmaceutical agent that improves the therapeutic effect of an immune checkpoint inhibitor when it is reused in a cancer patient after the administration of the immune checkpoint inhibitor has been discontinued.

[0005] The present invention encompasses the following inventions to solve the above-mentioned problems. [1] A pharmaceutical for improving the therapeutic effect of a second immune checkpoint inhibitor, the pharmaceutical comprising as an active ingredient an agent that promotes the expression of meflin, the pharmaceutical being administered in combination with a second immune checkpoint inhibitor to a cancer patient who has discontinued administration of a first immune checkpoint inhibitor. [2] The pharmaceutical according to [1] above, which is administered prior to administration of the second immune checkpoint inhibitor. [3] The pharmaceutical according to [1] or [2] above, wherein the second immune checkpoint inhibitor targets a molecule different from that of the first immune checkpoint inhibitor. [4] The pharmaceutical according to [1] or [2] above, wherein the second immune checkpoint inhibitor is a monoclonal antibody that targets the same molecule as the first immune checkpoint inhibitor but is of a different clone. [5] The pharmaceutical according to any of [1] to [4] above, wherein the agent that promotes the expression of meflin is a retinoid. [6] The pharmaceutical according to [5] above, wherein the retinoid is tamibarotene. [7] The pharmaceutical composition according to any one of [1] to [4] above, wherein the agent for promoting the expression of meflin is a composition for overexpressing meflin. [8] The pharmaceutical composition according to [7] above, wherein the composition for overexpressing meflin is a viral vector containing a meflin gene or a non-viral vector containing a meflin gene.

[0006] The present invention can provide a pharmaceutical agent that improves the therapeutic effect of an immune checkpoint inhibitor that has been reused in a cancer patient after the administration of the immune checkpoint inhibitor has been discontinued.

[0007] FIG. 1 is a diagram showing the protocol for Example 1. FIG. 1 is a diagram showing the results of measuring the tumor volume over time after transplantation in each group in Example 1, using mice into which mouse urothelial cancer cells were subcutaneously transplanted. FIG. 2 is a diagram showing the protocol for Example 2. FIG. 2 is a diagram showing the results of measuring the tumor volume over time after transplantation in each group in Example 2, using mice into which mouse urothelial cancer cells were subcutaneously transplanted. FIG. 3 is a diagram showing the protocol for Example 3. FIG. 3 is a diagram showing the results of measuring the tumor volume over time after transplantation in each group in Example 3, using mice into which mouse lung cancer cells were subcutaneously transplanted. FIG. 4 is a diagram showing the protocol for Example 5. FIG. 5 is a diagram showing the protocol for Example 6. FIG. 6 is a diagram showing the protocol for Example 7. FIG. 4 is a diagram showing the results of measuring the tumor volume over time after transplantation in each group in Example 7, using mice into which mouse urothelial cancer cells were subcutaneously transplanted.

[0008] The present invention provides a pharmaceutical for improving the therapeutic effect of a reused immune checkpoint inhibitor, which contains as an active ingredient an agent that promotes the expression of meflin. The pharmaceutical of the present invention is characterized in that it targets cancer patients who have discontinued administration of a first immune checkpoint inhibitor and is administered in combination with a second immune checkpoint inhibitor. The pharmaceutical of the present invention can improve the cancer therapeutic effect of the reused second immune checkpoint inhibitor.

[0009] The drug that promotes the expression of meflin, which is the active ingredient of the pharmaceutical of the present invention, may be a retinoid. The present inventors have confirmed that retinoids promote the expression of meflin in cancer-associated fibroblasts in the stroma of malignant tumors (WO2021 / 261601 A1). Examples of retinoids include acitretin, adapalene, AGN194204 (also referred to as IRX4204, NRX194204, or VTP 194204), AGN195183, alitretinoin (also referred to as 9-cis-retinoic acid), amsilarotene, AM580, bexarotene, bisphenol A diglycidyl ether (also referred to as BADGE), fenretinide, 4-hydroxyretinoic acid, isotretinoin (also referred to as 13-cis-retinoic acid), LG268, LGD1550, peretinoin, and retinyl acetate. acetate), tamibarotene (also known as AM80), tazarotene, tretinoin (also known as all-trans retinoic acid (ATRA)), TTNPB, etc. Preferred is Tamibarotene. As used herein, "retinoid" includes "vitamin A derivatives," "vitamin A analogs," "retinoic acid receptor (RAR) agonists," and "retinoid X receptor (RXR) agonists," but does not include "retinoic acid receptor (RAR) antagonists" or "retinoid X receptor (RXR) antagonists."

[0010] The retinoid may form a salt, and the salt is preferably a pharmaceutically acceptable salt. Examples include salts with acids such as hydrochloric acid, sulfuric acid, lactic acid, tartaric acid, maleic acid, fumaric acid, oxalic acid, malic acid, citric acid, oleic acid, palmitic acid, nitric acid, phosphoric acid, trifluoroacetic acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid; salts with hydroxides or carbonates of alkali metals or alkaline earth metals such as sodium, potassium, and calcium, or of aluminum; salts with triethylamine, benzylamine, diethanolamine, t-butylamine, dicyclohexylamine, and arginine.

[0011] The retinoid used in the medicament of the present invention can be formulated by appropriately blending pharmaceutically acceptable carriers or additives according to known methods for producing pharmaceutical preparations (e.g., methods described in the Japanese Pharmacopoeia, etc.). Specific examples include oral or parenteral preparations such as tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, buccal tablets, etc.), pills, powders, granules, capsules (including soft capsules and microcapsules), troches, syrups, liquids, emulsions, suspensions, controlled-release preparations (e.g., immediate-release preparations, sustained-release preparations, sustained-release microcapsules, etc.), aerosols, films (e.g., orally disintegrating films, oral mucosal patch films, etc.), injections (e.g., subcutaneous injections, intravenous injections, intramuscular injections, intraperitoneal injections, etc.), drip infusions, transdermal preparations, ointments, lotions, patches, suppositories (e.g., rectal suppositories, vaginal suppositories, etc.), pellets, nasal preparations, pulmonary preparations (inhalants), and eye drops. The blending ratio of the carrier or additive can be appropriately set based on the range usually adopted in the pharmaceutical field. The carrier or additive that can be blended is not particularly limited, and examples thereof include various carriers such as water, physiological saline, other aqueous solvents, aqueous or oily bases, and various additives such as excipients, binders, pH adjusters, disintegrants, absorption enhancers, lubricants, colorants, flavorings, and fragrances.

[0012] Examples of additives that can be incorporated into tablets, capsules, etc. include binders such as gelatin, cornstarch, tragacanth, and gum arabic; fillers such as crystalline cellulose; bulking agents such as cornstarch, gelatin, and alginic acid; lubricants such as magnesium stearate; sweeteners such as sucrose, lactose, or saccharin; and flavorings such as peppermint, saffron oil, and cherry. When the dosage unit is a capsule, the above-mentioned materials may further contain a liquid carrier such as an oil or fat. Sterile compositions for injection can be prepared according to standard formulation procedures (e.g., dissolving or suspending the active ingredient in a solvent such as water for injection or natural vegetable oil). Aqueous solutions for injection include, for example, physiological saline, isotonic solutions containing glucose or other adjuvants (e.g., D-sorbitol, D-mannitol, sodium chloride, etc.), and the like, which may be used in combination with appropriate solubilizing agents such as alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), and nonionic surfactants (e.g., polysorbate 80, HCO-50, etc.). As the oily liquid, for example, sesame oil, soybean oil, etc. are used, and may be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. Furthermore, the liquid may be blended with a buffer (e.g., phosphate buffer, sodium acetate buffer, etc.), a soothing agent (e.g., benzalkonium chloride, procaine hydrochloride, etc.), a stabilizer (e.g., human serum albumin, polyethylene glycol, etc.), a preservative (e.g., benzyl alcohol, phenol, etc.), an antioxidant, etc.

[0013] Retinoids have low toxicity and can be safely administered to humans and other mammals (e.g., rats, mice, rabbits, sheep, pigs, cows, cats, dogs, monkeys, etc.) The content of retinoid in the formulation varies depending on the dosage form, administration method, carrier, etc., but is usually 0.01 to 100% (w / w) of the total formulation, and may be 0.1 to 95% (w / w).

[0014] The dosage of retinoids varies depending on the subject, symptoms, route of administration, etc., but in the case of oral administration, for example, for a human weighing approximately 60 kg, it is approximately 0.01 to 1000 mg, preferably approximately 0.1 to 500 mg, and more preferably approximately 0.5 to 100 mg per day. In the case of parenteral administration, the single dose varies depending on the patient's condition, symptoms, administration method, etc., but for example, in the case of injections, it is usually administered intravenously at approximately 0.01 to 1000 mg, preferably approximately 0.01 to 500 mg, and more preferably approximately 0.01 to 20 mg per kg of body weight. The total daily dosage may be a single dose or divided doses.

[0015] The agent promoting the expression of meflin, the active ingredient of the pharmaceutical of the present invention, may be a composition that overexpresses meflin. The composition that overexpresses meflin is preferably a composition that overexpresses meflin in tumor tissue cells. Suitable compositions for overexpressing meflin include a plasmid vector, a viral vector, or the like, into which DNA encoding meflin has been inserted in an expressible manner. The nucleotide sequence of the gene encoding meflin can be easily obtained from known databases (e.g., NCBI). For example, the accession numbers for the nucleotide sequence of the human meflin gene (ISLR) are NM_201526.2 or NM_005545.4. Plasmid or viral vectors expressing meflin can be constructed using known genetic engineering techniques based on the obtained nucleotide sequence of the human meflin gene (ISLR).

[0016] When the composition for overexpressing meflin is administered in the form of a non-viral vector (such as a plasmid vector), methods that can be used include introducing a meflin expression plasmid using liposomes (such as the liposome method, HVJ-liposome method, cationic liposome method, lipofection method, and lipofectamine method), microinjection, and transferring a meflin expression plasmid to cells together with a carrier (metal particles) using a gene gun. When the composition is administered in the form of a viral vector, a meflin expression cassette can be introduced into a DNA or RNA virus such as a detoxified retrovirus, adenovirus, adeno-associated virus, herpesvirus, vaccinia virus, poxvirus, poliovirus, Sindbis virus, Sendai virus, or SV40, and the meflin gene can be introduced by infecting the target tumor with the viral vector.

[0017] The dosage of the composition that induces overexpression of meflin is preferably determined appropriately taking into consideration the type of vector, expression efficiency, tumor size, and the like.

[0018] The medicament of the present invention and the second immune checkpoint inhibitor may be administered to a subject simultaneously or at different times. As used herein, "administered in combination" means that the administration of the medicament of the present invention and the administration of the second immune checkpoint inhibitor overlap or are close to each other, and does not necessarily require simultaneous administration. It is preferable that administration of the medicament of the present invention be initiated prior to administration of the second immune checkpoint inhibitor, with the second immune checkpoint inhibitor being initiated at a later time. The time difference between the start of administration of the medicament of the present invention and the start of administration of the second immune checkpoint inhibitor is not particularly limited, and may be 1 day or more, 2 days or more, 3 days or more, 4 days or more, 5 days or more, 6 days or more, or 7 days or more. The administration period of the medicament of the present invention and the administration period of the second immune checkpoint inhibitor may overlap, or administration of the second immune checkpoint inhibitor may be initiated after the end of the administration period of the medicament of the present invention. The administration period and frequency of the medicament of the present invention are not particularly limited and may be determined appropriately by a physician.

[0019] In the present invention, the immune checkpoint inhibitor is not particularly limited. Target molecules of immune checkpoint inhibitors include, for example, CTLA-4, PD-1, LAG-3, BTLA, KIR, TIM-3, PD-L1, PD-L2, B7-H3, B7-H4, HVEM, GAL9, CD160, VISTA, BTNL2, TIGIT, PVR, BTN1A1, BTN2A2, BTN3A2, and CSF-1R. The target molecule of the immune checkpoint inhibitor may be CTLA-4, PD-1, or PD-L1. Monoclonal antibodies targeting human CTLA-4 include ipilimumab and tremelimumab, monoclonal antibodies targeting human PD-1 include nivolumab, pembrolizumab, spartalizumab, and cemiplimab, and monoclonal antibodies targeting human PD-L1 include avelumab, atezolizumab, and durvalumab.

[0020] The pharmaceutical of the present invention is intended for cancer patients in whom administration of a first immune checkpoint inhibitor has been discontinued. The first immune checkpoint inhibitor refers to an immune checkpoint inhibitor that has been administered to a cancer patient for the first time. The reason for discontinuing administration of the first immune checkpoint inhibitor is not particularly limited. Examples include discontinuing administration of the first immune checkpoint inhibitor due to immune checkpoint inhibitor resistance caused by administration of the first immune checkpoint inhibitor, discontinuing administration of the first immune checkpoint inhibitor because it is ineffective, or discontinuing administration due to side effects caused by administration of the first immune checkpoint inhibitor. The period during which the first immune checkpoint inhibitor is discontinued, i.e., the timing to start administration of the second immune checkpoint inhibitor, is not particularly limited and is determined appropriately by a physician.

[0021] The second immune checkpoint inhibitor preferably targets a molecule different from that of the first immune checkpoint inhibitor. That is, for example, if the target molecule of the first immune checkpoint inhibitor is PD-1, the second immune checkpoint inhibitor preferably targets a molecule other than PD-1 (e.g., PD-L1, CTLA-4, etc.), if the target molecule of the first immune checkpoint inhibitor is PD-L1, the second immune checkpoint inhibitor preferably targets a molecule other than PD-L1 (e.g., PD-1, CTLA-4, etc.), and if the target molecule of the first immune checkpoint inhibitor is CTLA-4, the second immune checkpoint inhibitor preferably targets a molecule other than CTLA-4 (e.g., PD-1, PD-L1, etc.).

[0022] The second immune checkpoint inhibitor may be a monoclonal antibody that targets the same molecule as the first immune checkpoint inhibitor but is a different clone. For example, if the target molecule is human PD-1 and the first immune checkpoint inhibitor is nivolumab, the second immune checkpoint inhibitor may be pembrolizumab, spartalizumab, cemiplimab, or the like. For example, if the target molecule is human PD-L1 and the first immune checkpoint inhibitor is avelumab, the second immune checkpoint inhibitor may be atezolizumab, durvalumab, or the like. For example, if the target molecule is human CTLA-4 and the first immune checkpoint inhibitor is ipilimumab, the second immune checkpoint inhibitor may be tremelimumab, or the like. The combination of the first immune checkpoint inhibitor and the second immune checkpoint inhibitor is not limited to the above.

[0023] The present invention includes the following inventions: (1) A method for treating cancer, comprising administering an agent that promotes the expression of meflin in combination with a second immune checkpoint inhibitor to a cancer patient who has discontinued administration of a first immune checkpoint inhibitor. (2) A method for enhancing the therapeutic effect of a second immune checkpoint inhibitor in a cancer patient who has discontinued administration of a first immune checkpoint inhibitor, comprising administering an agent that promotes the expression of meflin in combination with the second immune checkpoint inhibitor to a cancer patient who has discontinued administration of the first immune checkpoint inhibitor. (3) An agent that promotes the expression of meflin, used to enhance the therapeutic effect of a second immune checkpoint inhibitor in a cancer patient who has discontinued administration of the first immune checkpoint inhibitor. (4) Use of an agent that promotes the expression of meflin for manufacturing a medicament for improving the therapeutic effect of a second immune checkpoint inhibitor in a cancer patient who has discontinued administration of the first immune checkpoint inhibitor.

[0024] The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

[0025] Example 1: Effect of Tamibarotene in Combination with Anti-PD-L1 Antibody or Anti-PD-1 after Anti-PD-L1 Antibody Treatment on Mouse Urothelial Cancer Cells (1) Experimental Method: C57BL / 6J female mice were inoculated with mouse urothelial cancer cell MB49 (1.0 × 10 6(100 pieces) were subcutaneously implanted. Anti-PD-L1 antibody (250 μg) was administered on days 9, 12, and 15. On day 21, the tumors were divided into six groups (n=5 per group): control (DMSO + control group), tamibarotene (AM80 + control group), anti-PD-L1 antibody (DMSO + αPD-L1 group), anti-PD-1 antibody (DMSO + αPD-1 group), anti-PD-L1 antibody and tamibarotene (AM80 + αPD-L1 group), and anti-PD-1 antibody and tamibarotene (AM80 + αPD-1 group) to equalize tumor volume. The AM80 + control group, AM80 + αPD-L1 group, and AM80 + αPD-1 group received oral tamibarotene (3.0 mg / kg) once daily for 7 days, starting on day 21. The DMSO + control group, DMSO + αPD-L1 group, and DMSO + αPD-1 group will be orally administered DMSO instead of tamibarotene. The DMSO + αPD-L1 group and AM80 + αPD-L1 group will be intraperitoneally administered anti-PD-L1 antibody (250 μg) on ​​days 28 and 31. The DMSO + αPD-1 group and AM80 + αPD-1 group will be intraperitoneally administered anti-PD-1 antibody (200 μg) on ​​days 28 and 31. The DMSO + control group and AM80 + control group will be intraperitoneally administered isotype IgG instead of anti-PD-L1 antibody or anti-PD-1 antibody on days 28 and 31. The protocol is shown in Figure 1.

[0026] (2) Results The results are shown in Figure 2. Figure 2 shows that when resistance developed after first-line anti-PD-L1 antibody administration, significant tumor growth suppression was observed only when anti-PD-1 antibody was administered after oral administration of Am80. This result indicates that for AM80 to have an ICI sensitivity-enhancing effect in second-line treatment, the type of ICI used in second-line treatment must be different from that used in first-line treatment.

[0027] Example 2: Effect of Tamibarotene in Combination with Anti-PD-L1 Antibody or Anti-PD-1 on Mouse Urothelial Cancer Cells after Anti-PD-1 Antibody Treatment (1) Experimental Method: C57BL / 6J female mice were inoculated with mouse urothelial cancer cell MB49 (1.0 × 10 6(100 pieces) were subcutaneously implanted. Anti-PD-1 antibody (200 μg) was administered on days 9, 12, and 15. On day 21, the tumors were divided into six groups (n=5 per group): control (DMSO + control group), tamibarotene (AM80 + control group), anti-PD-L1 antibody (DMSO + αPD-L1 group), anti-PD-1 antibody (DMSO + αPD-1 group), anti-PD-L1 antibody and tamibarotene (AM80 + αPD-L1 group), and anti-PD-1 antibody and tamibarotene (AM80 + αPD-1 group) to equalize tumor volume. The AM80 + control group, AM80 + αPD-L1 group, and AM80 + αPD-1 group received oral tamibarotene (3.0 mg / kg) once daily for 7 days, starting on day 21. The DMSO + control group, DMSO + αPD-L1 group, and DMSO + αPD-1 group will be orally administered DMSO instead of tamibarotene. The DMSO + αPD-L1 group and AM80 + αPD-L1 group will be intraperitoneally administered anti-PD-L1 antibody (250 μg) on ​​days 28 and 31. The DMSO + αPD-1 group and AM80 + αPD-1 group will be intraperitoneally administered anti-PD-1 antibody (200 μg) on ​​days 28 and 31. The DMSO + control group and AM80 + control group will be intraperitoneally administered isotype IgG instead of anti-PD-L1 antibody or anti-PD-1 antibody on days 28 and 31. The protocol is shown in Figure 3.

[0028] (2) Results The results are shown in Figure 4. Figure 4 shows that when resistance developed after first-line anti-PD-1 antibody administration, significant tumor growth suppression was observed only when anti-PD-L1 antibody was administered after oral administration of Am80. This result indicates that for AM80 to have an ICI sensitivity-enhancing effect in second-line treatment, the type of ICI used in second-line treatment must be different from that used in first-line treatment.

[0029] Example 3: Effect of Tamibarotene in Combination with Anti-PD-L1 Antibody or Anti-PD-1 after Anti-PD-L1 Antibody Treatment on Mouse Lung Cancer Cells (1) Experimental Method: Mouse lung cancer cells LLC-luc (1.0 × 10 6(100 pieces) were subcutaneously implanted. Anti-PD-L1 antibody (250 μg) was administered on days 9, 12, and 15. On day 21, the tumors were divided into six groups (n=5 per group): control (DMSO + control group), tamibarotene (AM80 + control group), anti-PD-L1 antibody (DMSO + αPD-L1 group), anti-PD-1 antibody (DMSO + αPD-1 group), anti-PD-L1 antibody and tamibarotene (AM80 + αPD-L1 group), and anti-PD-1 antibody and tamibarotene (AM80 + αPD-1 group) to equalize tumor volume. The AM80 + control group, AM80 + αPD-L1 group, and AM80 + αPD-1 group received oral tamibarotene (3.0 mg / kg) once daily for 7 days, starting on day 21. The DMSO + control group, DMSO + αPD-L1 group, and DMSO + αPD-1 group were orally administered DMSO instead of tamibarotene. The DMSO + αPD-L1 group and AM80 + αPD-L1 group were intraperitoneally administered anti-PD-L1 antibody (250 μg) on ​​days 28, 31, and 34. The DMSO + αPD-1 group and AM80 + αPD-1 group were intraperitoneally administered anti-PD-1 antibody (200 μg) on ​​days 28, 31, and 34. The DMSO + control group and AM80 + control group were intraperitoneally administered isotype IgG instead of anti-PD-L1 antibody or anti-PD-1 antibody on days 28, 31, and 34. The protocol is shown in Figure 5.

[0030] (2) Results The results are shown in Figure 6. Figure 6 shows that when resistance developed after first-line anti-PD-L1 antibody administration, oral administration of Am80 followed by an anti-PD-1 antibody, an ICI different from the one used in first-line treatment, significantly suppressed tumor growth. This result indicates that for AM80 to have an ICI-enhancing effect on second-line treatment, the ICI used in second-line treatment must be different from the ICI used in first-line treatment.

[0031] Example 4: Effect of Tamibarotene in Combination with Anti-PD-L1 Antibody or Anti-PD-1 on Mouse Lung Cancer Cells after Anti-PD-1 Antibody Treatment (1) Experimental Method: Mouse lung cancer cells LLC-luc (1.0 × 10 6(100 pieces) were subcutaneously implanted. Anti-PD-1 antibody (200 μg) was administered on days 9, 12, and 15. On day 21, the tumors were divided into six groups (n=5 per group): control (DMSO + control group), tamibarotene (AM80 + control group), anti-PD-L1 antibody (DMSO + αPD-L1 group), anti-PD-1 antibody (DMSO + αPD-1 group), anti-PD-L1 antibody and tamibarotene (AM80 + αPD-L1 group), and anti-PD-1 antibody and tamibarotene (AM80 + αPD-1 group) to equalize tumor volume. The AM80 + control group, AM80 + αPD-L1 group, and AM80 + αPD-1 group received oral tamibarotene (3.0 mg / kg) once daily for 7 days, starting on day 21. The DMSO + control group, DMSO + αPD-L1 group, and DMSO + αPD-1 group were orally administered DMSO instead of tamibarotene. The DMSO + αPD-L1 group and AM80 + αPD-L1 group received intraperitoneal administration of anti-PD-L1 antibody (250 μg) on ​​days 28, 31, and 34. The DMSO + αPD-1 group and AM80 + αPD-1 group received intraperitoneal administration of anti-PD-1 antibody (200 μg) on ​​days 28, 31, and 34. The DMSO + control group and AM80 + control group received intraperitoneal administration of isotype IgG instead of anti-PD-L1 antibody or anti-PD-1 antibody on days 28, 31, and 34. The protocol is shown in Figure 7.

[0032] (2) Results The results are shown in Figure 8. Figure 8 shows that when resistance developed after first-line anti-PD-1 antibody administration, oral administration of Am80 followed by an anti-PD-L1 antibody, an ICI different from the one used in first-line treatment, significantly suppressed tumor growth. This result indicates that for AM80 to have an ICI-enhancing effect on second-line ICIs, the type of ICI used in second-line treatment must be different from that used in first-line treatment.

[0033] Example 5: Changes caused by resistance in mouse urothelial cancer cells after anti-PD-L1 antibody or anti-PD-1 antibody treatment and mechanism of overcoming the resistance by tamibarotene administration (1) Experimental method: C57BL / 6J female mice were inoculated with mouse urothelial cancer cells MB49 (1.0 × 106 Mice were then subcutaneously implanted with αPD-L1 antibodies (DMSO + αPD-L1 group), anti-PD-1 antibodies (DMSO + αPD-1 group), anti-PD-L1 antibodies and tamibarotene (AM80 + αPD-L1 group), and anti-PD-1 antibodies and tamibarotene (AM80 + αPD-1 group). On days 9, 12, and 15, the DMSO + αPD-L1 and AM80 αPD-L1 groups received 250 μg of anti-PD-L1 antibodies, while the DMSO + αPD-1 and AM80 + αPD-1 groups received 200 μg of anti-PD-1 antibodies. Furthermore, the AM80 + αPD-1 and AM80 + αPD-L1 groups received oral tamibarotene (3.0 mg / kg) once daily for 7 days starting on Day 21, while the DMSO + αPD-1 and DMSO + αPD-L1 groups received oral DMSO. Tumors were removed from these four groups on Day 28, and immunohistochemistry (IHC), single-cell RNA sequencing, and spatial transcriptome analysis were used to elucidate the mechanisms underlying the overcoming of ICI resistance following tamibarotene administration. The protocol is shown in Figure 9.

[0034] Example 6: Changes caused by resistance in mouse lung cancer cells after anti-PD-L1 antibody or anti-PD-1 antibody treatment and mechanism of overcoming the resistance by tamibarotene administration (1) Experimental method Mouse lung cancer cells LLC-luc (1.0 × 10 6Mice were then subcutaneously implanted with αPD-L1 antibodies (DMSO + αPD-L1 group), anti-PD-1 antibodies (DMSO + αPD-1 group), anti-PD-L1 antibodies and tamibarotene (AM80 + αPD-L1 group), and anti-PD-1 antibodies and tamibarotene (AM80 + αPD-1 group). On days 9, 12, and 15, the DMSO + αPD-L1 and AM80 αPD-L1 groups received 250 μg of anti-PD-L1 antibodies, while the DMSO + αPD-1 and AM80 + αPD-1 groups received 200 μg of anti-PD-1 antibodies. Furthermore, the AM80 + αPD-1 and AM80 + αPD-L1 groups received oral tamibarotene (3.0 mg / kg) once daily for 7 days starting on Day 21, while the DMSO + αPD-1 and DMSO + αPD-L1 groups received oral DMSO. Tumors were removed from these four groups on Day 28, and immunohistochemistry (IHC), single-cell RNA sequencing, and spatial transcriptome analysis were used to elucidate the mechanisms underlying the overcoming of ICI resistance following tamibarotene administration. The protocol is shown in Figure 10.

[0035] Example 7: Effect of combined use of tamibarotene and a second anti-PD-1 antibody on mouse urothelial cancer cells after a first anti-PD-1 antibody treatment (1) Experimental method C57BL / 6J female mice were inoculated with mouse urothelial cancer cells MB49 (1.0 × 10 6Anti-PD-1 antibody A (clone name: 29F.1A12) (200 μg) was administered on days 9, 12, and 15, and on day 21, the mice were divided into six groups (n=5 per group) to ensure equal tumor volume: control group (DMSO + control group), tamibarotene group (AM80 + control group), anti-PD-1 antibody A group (DMSO + αPD-1A group), anti-PD-1 antibody B (clone name: RMP1-14) group (DMSO + αPD-1B group), anti-PD-1 antibody A and tamibarotene group (AM80 + αPD-1A group), and anti-PD-1 antibody B and tamibarotene group (AM80 + αPD-1B group). The AM80 + control, AM80 + αPD-1A, and AM80 + αPD-1B groups received oral tamibarotene (3.0 mg / kg) once daily for 7 days starting on Day 21. The DMSO + control, DMSO + αPD-1A, and DMSO + αPD-1B groups received oral DMSO instead of tamibarotene. The DMSO + αPD-1A and AM80 + αPD-1A groups received intraperitoneal administration of anti-PD-1 antibody A (200 μg) on ​​Days 28 and 31. The DMSO + αPD-1B and AM80 + αPD-1B groups received intraperitoneal administration of anti-PD-1 antibody B (200 μg) on ​​Days 28 and 31. The DMSO + control and AM80 + control groups received intraperitoneal administration of isotype IgG instead of anti-PD-1 antibody on Days 28 and 31. The protocol is shown in FIG.

[0036] (2) Results The results are shown in Figure 12. Figure 12 shows that when resistance developed after administration of anti-PD-1 antibody A as first-line therapy, only oral administration of Am80 followed by administration of anti-PD-1 antibody B of a different clone significantly suppressed tumor growth. This result indicates that for AM80 to have an effect of enhancing the sensitivity of second-line ICI therapy, the second-line ICI can be an antibody targeting the same molecule as the ICI used in first-line therapy, as long as it is a different clone.

[0037] The present invention is not limited to the above-described embodiments and examples, and various modifications are possible within the scope of the claims. The technical scope of the present invention also includes embodiments obtained by appropriately combining the technical means disclosed in different embodiments. Furthermore, all academic literature and patent documents described in this specification are incorporated herein by reference.

Claims

1. A pharmaceutical for improving the therapeutic effect of a second immune checkpoint inhibitor, the pharmaceutical containing as an active ingredient an agent that promotes the expression of meflin, which is administered in combination with a second immune checkpoint inhibitor to cancer patients who have discontinued administration of a first immune checkpoint inhibitor.

2. The pharmaceutical according to claim 1, which is administered prior to the administration of a second immune checkpoint inhibitor.

3. The pharmaceutical described in claim 1 or 2, wherein the second immune checkpoint inhibitor targets a molecule different from that of the first immune checkpoint inhibitor.

4. The pharmaceutical composition of claim 1 or 2, wherein the second immune checkpoint inhibitor is a monoclonal antibody that targets the same molecule as the first immune checkpoint inhibitor but is of a different clone.

5. The pharmaceutical composition according to claim 1, wherein the drug that promotes the expression of meflin is a retinoid.

6. The pharmaceutical composition according to claim 5, wherein the retinoid is tamibarotene.

7. The pharmaceutical composition according to claim 1, wherein the agent that promotes the expression of meflin is a composition that causes the overexpression of meflin.

8. The pharmaceutical composition according to claim 7, wherein the composition for overexpressing meflin is a viral vector containing the meflin gene or a non-viral vector containing the meflin gene.

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