Serotonin receptors as therapeutic targets

Abstract

Serotonin receptor 5-HT for use in treating a proliferative disorder in a subject in need thereof 2A Inhibitors or serotonin receptor 5-HT 2A Also provided are compositions comprising an inhibitor of serotonin receptor 5-HT 2 receptors in the manufacture of a medicament for treating a proliferative disorder in a subject. 2A Uses of inhibitors and methods of treatment thereof are also disclosed. 2A Pharmaceutical compositions or combination therapies comprising the inhibitors and immune checkpoint blockade / ICB therapy are also disclosed.

Description

[Technical Field]

[0001] The present disclosure relates generally to methods of treating proliferative disorders. Specifically, the present disclosure relates to methods of treating 5-HT 2A It relates to serotonin receptor inhibitors. [Background technology]

[0002] Hepatocellular carcinoma (HCC) is the most common form of liver cancer (resulting in over 800,000 deaths worldwide annually), with the greatest disease burden in the Asia-Pacific region. In Singapore, HCC is the fourth most common cancer in men, with an age-standardized incidence rate of 17.7 cases per 100,000 men. While early-stage HCC can be treated by resection, local ablation therapy, or liver transplantation, 60–80% of such cases experience tumor recurrence within five years, often progressing to advanced HCC. Advanced HCC (aHCC) has a poor prognosis (median survival <1 year) and limited treatment options. Combination immunotherapy using anti-PD-L1 and anti-VEGFA monoclonal antibodies recently achieved promising breakthroughs, surpassing the efficacy of the historical systemic therapy sorafenib in the IMbrave150 clinical trial. However, considerable room for improvement remains, as two-thirds of aHCC patients remain ineffective to any type of treatment.

[0003] Furthermore, the molecular pathways targeted by all approved treatments for aHCC are very narrow (including the VEGF, PD-1, and CTLA-4 pathways). Therefore, there is a clinical need to explore and evaluate more potential molecular targets that can be rationally combined with existing approved immunotherapeutic drugs. Given that all three FDA-approved drugs for first-line HCC treatment were first approved for use in other non-HCC indications and then repurposed for HCC after successful clinical trials, there is a particular lack of HCC-specific therapeutics that target the liver immune microenvironment to improve therapeutic efficacy. Summary of the Invention [Problem to be solved by the invention]

[0004] Therefore, there is a need to provide alternative targeted treatments for proliferative diseases such as hepatocellular carcinoma. [Means for solving the problem]

[0005] In a first aspect, a serotonin receptor 5-HT agonist is provided for use in treating a proliferative disorder in a subject in need thereof. 2A Inhibitors or serotonin receptor 5-HT 2A Compositions comprising the inhibitors are provided.

[0006] In a second aspect, a method for treating a proliferative disorder in a subject in need thereof, comprising administering to a subject a serotonin receptor 5-HT agonist, a serotonin receptor agonist, a 5-HT ... 2A Use of an inhibitor is provided.

[0007] In a third aspect, a method of treating a proliferative disorder in a subject in need thereof comprises administering an effective amount of a serotonin receptor 5-HT 2A A method is provided that includes administering an inhibitor to a subject.

[0008] In some instances, the serotonin receptor 5-HT 2A The inhibitor improves suppression of the immune system, and optionally improves suppression of CD8 T cell function.

[0009] In some instances, 5-HT 2A The inhibitor is 5-HT 2A It is a selective antagonist of

[0010] In some instances, 5-HT 2A Inhibitors inhibit the serotonin receptor 5-HT by blocking its function in enhancing CD8 T cell activation 2A Agents targeting the 5-HT and / or serotonin receptors 2A and thus inhibit its function in enhancing CD8 T cell activation.

[0011] In some instances, 5-HT 2A The inhibitor is a compound having the formula: [ka] and pharmaceutically acceptable acid addition salts thereof and possible stereoisomers thereof, wherein R is hydrogen or C 1~6 is alkyl, Alk is C 1~4 is an alkanediyl, Q is a group of the following formula [ka] (In the formula, Y 1 and Y 2 are each independently O or S, R 2 is hydrogen, halo, C 1~6 Alkyl or C 1~6 alkyloxy, R 3 is hydrogen or halo) or Q is a group of the following formula [ka] (In the formula, R 4 is hydrogen or C 1~6 is alkyl, Z is -S-, -CH2- or -CR 5 =CR 6 wherein R 5 and R 6 are each independently hydrogen or C 1~6 alkyl, and A is a divalent group -CH2-CH2-, -CH2-CH2-CH2- or -CR 7 =CR 8 wherein R 7 and R 8 are each independently hydrogen, halo, amino or C 1~6 alkyl), R 1is the base of the following formula -X-Ar (c) wherein Ar is phenyl or substituted phenyl, said substituted phenyl having an amino group and / or 1, 2 or 3 halo atoms; X is >C=O, >CH-OH, >CH-OC(O)-R 9 , >CH2, >C(OC 1~6 alkyl)2, [ka] , >C=N-OH or >C=N-NH2; R 9 is hydrogen or C 1~6 is alkyl, wherein q is an integer of 2 or 3. or R 1 is the base of the following formula [ka] (In the formula, R 10 is hydrogen or C 1~6 alkyl, and R 11 , R 12 and R 13 are each independently hydrogen or halo. or R 1 is the base of the following formula [ka] (Wherein A is O or S, R 14 and R 15 are each independently hydrogen, halo, hydroxy, C 1~6 Alkyloxy or C 1~6 alkyl).

[0012] In some instances, 5-HT 2A The inhibitor is a compound having the formula: [ka] [In the formula, Z is [ka] R is hydrogen, a cyclic or linear or branched acyclic organyl group, a lower hydroxyalkyl group, a lower aminoalkyl group, or an aralkyl or heteroaralkyl group; n is 0, 1, or 2; X1 is methylene, vinylene, or an NH or N(lower alkyl) group; X2 is methylene, or when X1 is methylene or vinylene, X2 is methylene or a bond, or when X1 is methylene, X2 is O, S, NH, or N(lower alkyl) or a bond; Y1 is methylene and Y2 is methylene, vinylene, ethylene, propylene, or a bond; or Y1 is a bond and Y2 is vinylene, or Y1 is ethylene and Y2 is O, S, NH, or N(lower alkyl); Ar1 and Ar2 are independently an unsubstituted or substituted aryl or heteroaryl group; W is oxygen or sulfur; or It is a pharmaceutically acceptable salt, ester, or prodrug thereof.

[0013] In some instances, 5-HT 2A The inhibitor is a compound having the formula: [ka] wherein one of X and Y is CH2 and the other is selected from the group consisting of CH2, O, and S; The dotted line emanating from Z indicates an optional bond; when no bond is shown, Z is N, CH or COH; when a bond is shown, Z is C; Ar is phenyl, 2-thienyl, 3-thienyl, 2-furanyl, 3-furanyl, 2-pyrimidyl, 1-indolyl, 2-indolyl, 3-indolyl, 1-indol-2-onyl, 3-indol-2-onyl, 2- or 3-benzofuranyl, 2- or 3-benzothiophenyl, 1-naphthyl or 2-naphthyl, each optionally being halogen, lower alkyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, trifluoromethylsulfonyloxy, cycloalkyl, cycloalkyl-lower alkyl, nitro, amino, lower alkylamino, di-lower alkylamino, acylamino or C 1~2 substituted with alkylenedioxy; R 1 is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, cycloalk(en)yl, cycloalk(en)yl-lower alk(en / yn)yl, aryl, aryl-lower alkyl, acyl, thioacyl, lower alkylsulfonyl, trifluoromethylsulfonyl, arylsulfonyl, R 1 is R 9 VCO- group (wherein V is O or S and R 9 is lower alkyl, cycloalkyl, cycloalkyl lower alkyl, or aryl), or R 1 is R 10 R 11 NCO-group or R 10 R 11 NCS- group (wherein R 10 and R 11 are independently hydrogen, lower alkyl, cycloalkyl, cycloalkyl-lower alkyl, or aryl, or R 10 and R 11 together with the N atom to which they are attached form a pyrrolidinyl, piperidinyl or perhydroazepine group, R 2 is hydrogen, lower alkyl, cycloalkyl, or cycloalkyl-lower alkyl; or R 1 and R 2together with the N atom to which they are attached. [ka] wherein Q is C=O, C=S or CH, T is NH, S, O or CH, and m is 1 to 4 inclusive; R 3 ~R 5 are independently hydrogen, halogen, lower alkyl, lower alkylcarbonyl, phenylcarbonyl, halogen-substituted phenylcarbonyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, cycloalkyl, cycloalkyl-lower alkyl, or nitro; R 6 and R 7 are each hydrogen or lower alkyl, or they are joined together to form a 3- to 7-membered carbocyclic ring; R 8 is hydrogen or lower alkyl, any alkyl, cycloalkyl or cycloalkylalkyl groups present are optionally substituted with one or two hydroxy groups, which in turn are optionally esterified with an aliphatic or aromatic carboxylic acid, and any aryl substituents present are optionally substituted with halogen, lower alkyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, trifluoromethylsulfonyloxy, cycloalkyl, cycloalkyl-lower alkyl or nitro, and pharmaceutically acceptable acid addition salts thereof.

[0014] In some instances, 5-HT 2AThe inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate), 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound.

[0015] In some instances, the proliferative disorder is a tumor and / or cancer.

[0016] In some instances, the proliferative disease is liver cancer.

[0017] In some instances, the proliferative disease is hepatocellular carcinoma.

[0018] In some examples, the present invention relates to a method for treating serotonin receptor 5-HT2+ receptor agonists, including the use of the present invention, which further comprises one or more combination therapies selected from the group consisting of immunotherapy, chemotherapy, ablation therapy, and transplantation. 2A 10. The method of claim 2, wherein the inhibitor is a compound of formula (I) or (II) as defined herein.

[0019] In some examples, the serotonin receptor 5-HT2 receptor agonist for the uses described herein further includes immunotherapy, such as, but not limited to, immune checkpoint blockade (ICB) therapy, including administration of a monoclonal antibody, and optionally, the immunotherapy may include the use of one or more of anti-PD-L1 (such as atezolizumab or nivolumab), anti-VEGFA (such as bevacizumab), anti-CTLA4 (such as ipilimumab), or anti-PD-1 (such as nivolumab) plus anti-CTLA4 (such as ipilimumab), and / or combinations thereof. 2A An inhibitor or a use as described herein or a method as described herein.

[0020] In some instances, the combination therapy inhibits the serotonin receptor 5-HT 2A The inhibitors may be administered to the subject simultaneously, sequentially, or separately.

[0021] In some instances, the inhibitor is to be administered intravenously, subcutaneously, orally, sublingually, or intraperitoneally.

[0022] In a fourth aspect, the serotonin receptor 5-HT 2A Pharmaceutical compositions comprising the inhibitors and immune checkpoint blockade / ICB therapy are provided.

[0023] In some examples, the serotonin receptor inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate), 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound.

[0024] In a fifth aspect, the serotonin receptor 5-HT 2A and combination therapy including immune checkpoint blockade / ICB therapy, wherein the serotonin receptor inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate). , 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound.

[0025] definition As used herein, the term "halo" refers to fluoro, chloro, bromo, and iodo, and "C 1~6 The term "alkyl" refers to straight and branched chain saturated hydrocarbon groups having 1 to 6 carbon atoms, e.g., methyl, ethyl, 1-methylethyl, 1,1-dimethylethyl, propyl, 2-methylpropyl, butyl, pentyl, hexyl, and the like, and "C 1~4 "Alkanediyl" includes divalent straight or branched chain alkanediyl groups having 1 to 4 carbon atoms.

[0026] As used herein, the term "inhibitor" or "antagonist" refers to an agent that reduces the amount or duration of the effect of the biological activity of serotonin, particularly at the serotonin receptor 5-HT 2A In some instances, inhibitors or antagonists may include compounds, proteins, nucleic acids, carbohydrates, antibodies, or small molecules that decrease the effects of serotonin. For example, antagonists may decrease the effects of serotonin receptor 5-HT 2Acan be used in accordance with the present disclosure to inhibit

[0027] The terms "treating," "treat," and "therapy," and their synonyms, refer to both therapeutic treatment and prophylactic or preventative measures aimed at preventing or slowing (alleviating) medical conditions, including, but not limited to, diseases (such as proliferative diseases, including tumors and / or cancers), symptoms, and disorders. Medical conditions also include the body's response to a disease or disorder, such as dysregulated cell proliferation, dysregulated cell metabolism, and / or inflammation. Those in need of such treatment include those already with a medical condition and those prone to developing a medical condition or in whom a medical condition is to be prevented. In some examples, the treatments described herein slow tumor growth and / or prolong survival. In some examples, the present disclosure may result in higher survival rates, slowed tumor growth, and / or lower mortality rates. In some examples, the present disclosure, when used alone or in combination with therapies known in the art (e.g., immune checkpoint blockade (ICB) therapy), may result in higher survival rates, delayed tumor growth, and / or lower mortality rates.

[0028] As used herein, the term "subject" includes patients and non-patients. The term "patient" refers to an individual who is suffering from or likely to suffer from a medical condition, such as a proliferative disease, including tumors and / or cancer, while "non-patient" refers to an individual who is not suffering from or likely to not suffer from a medical condition. "Non-patient" includes healthy individuals, individuals who are not affected by the disease, and / or individuals who are free of the medical condition. The term "subject" includes humans and animals. Animals include, but are not limited to, mammals (e.g., non-human primates, dogs, mice, rabbits, etc.) and the like. "Mouse" refers to any mammal of the murine family, such as mice and rats.

[0029] The term "and / or," e.g., "X and / or Y," should be understood to mean either "X and Y" or "X or Y," and should be interpreted as clearly endorsing both meanings or either meaning.

[0030] Furthermore, in the description herein, whenever the word "substantially" is used, it is understood to include, but not be limited to, "entirely" or "completely." In addition, terms such as "comprising," "comprise," and the like, whenever used, are intended to be open-ended descriptive language in that they broadly include the elements / components listed after the term, in addition to other components not explicitly listed. For example, when "comprising" is used, a reference to "one" feature is also intended to be a reference to "at least one" of that feature. Terms such as "consisting," "consist," and the like may, in appropriate context, be considered subsets of terms such as "comprising," "comprise," and the like. Thus, in embodiments disclosed herein using terms such as "comprising," "comprise," and the like, it will be understood that these embodiments also provide teachings of corresponding embodiments using terms such as "consisting," "consist," and the like. Furthermore, terms such as "about," "approximately," and the like, whenever used, typically refer to a reasonable variation, such as a + / - 5% variation from the disclosed value, or a 4% variation from the disclosed value, or a 3% variation from the disclosed value, or a 2% variation from the disclosed value, or a 1% variation from the disclosed value.

[0031] Furthermore, in the present description, certain values ​​may be disclosed in ranges. The endpoints of the ranges are intended to indicate preferred ranges. Whenever a range is described, it is intended that the range encompass and teach all possible subranges and individual values ​​within that range. That is, the endpoints of the range should not be construed as inflexible limitations. For example, reciting a range of 1% to 5% is intended to have specifically disclosed subranges of 1% to 2%, 1% to 3%, 1% to 4%, 2% to 3%, etc., as well as individually values ​​within that range such as 1%, 2%, 3%, 4%, and 5%. It should be understood that the individual values ​​within a range include integers, fractions, and decimals. Furthermore, whenever a range is described, it is also intended that the range encompass and teach values ​​from the endpoints recited up to two additional decimal places or significant digits (where appropriate). For example, reciting a range of 1% to 5% is intended to have the specifically disclosed range of 1.00% to 5.00%, as well as 1.0% to 5.0%, and all intermediate values ​​therein (1.01%, 1.02%, 4.98%, 4.99%, 5.00%, and 1.1%, 1.2%, 4.8%, 4.9%, 5.0%, etc.) The above specific disclosure intent is applicable to any depth / breadth of range.

[0032] Additionally, when describing some embodiments, the present disclosure may disclose a method and / or process as having steps in a particular order. However, unless otherwise specified, the method or process should not be limited to the particular order of steps disclosed. Other orders of steps may be possible. The particular order of steps disclosed herein should not be construed as an undue limitation. Unless otherwise specified, the method and / or process disclosed herein should not be limited to steps performed in the order described. The order of steps may be changed and still remain within the scope of the present disclosure.

[0033] Furthermore, although the present disclosure provides embodiments having one or more of the features / characteristics discussed herein, it is understood that one or more of these features / characteristics may be disclaimed in other alternative embodiments, and that the present disclosure supports such disclaimers and these related alternative embodiments. DETAILED DESCRIPTION OF THE INVENTION

[0034] Serotonin receptor 5-HT 2A Exemplary, non-limiting embodiments of inhibitors and methods of treatment using the inhibitors are disclosed below.

[0035] 5-HT, one of three members of the 5-HT2 receptor family 2A The receptor is a typical seven-transmembrane G protein receptor that couples to the Gq / 11 signaling protein. 2A The receptors are present postsynaptically on serotonergic neurons. 2A The receptors are primarily found in the frontal cortex or in areas connected to the visual cortex. 2A The receptor can be found to elicit a response in vascular smooth muscle and a component of the response of gastrointestinal smooth muscle.

[0036] The inventors of the present disclosure have demonstrated that serotonin receptor 5-HT 2A The inventors of the present disclosure have discovered that 5-HT2 receptors have a surprising inhibitory effect on hepatic NPC-activated CD8 T cells. 2A We found that this could be a surprisingly effective target for adjuvant therapy with HCC immunotherapy, potentially fulfilling an important clinical need for HCC-specific molecular therapies that can be rationally combined with existing approved immunotherapeutics.

[0037] The liver immune microenvironment is inherently immunosuppressive because hepatocytes and nonparenchymal cells (NPCs), such as Kupffer cells and sinusoidal endothelial cells, suppress T cell activation under normal resting conditions. Therefore, inhibiting these immunosuppressive interactions between hepatic NPCs and tumor-specific T cells could potentially enhance the efficacy of T cell-targeted immunotherapy in HCC.

[0038] As such, in one aspect, a serotonin receptor 5-HT agonist is provided for use in treating a proliferative disorder in a subject in need thereof. 2A Inhibitors or serotonin receptor 5-HT 2A Compositions comprising the inhibitors are provided.

[0039] In another aspect, a method for treating a proliferative disorder in a subject in need thereof, comprising administering to a subject an active ingredient in a serotonin receptor 5-HT agonist ... 2A Use of an inhibitor is provided.

[0040] In yet another aspect, a method of treating a proliferative disorder in a subject in need thereof includes administering an effective amount (e.g., a therapeutically effective amount) of a serotonin receptor 5-HT 2A A method is provided that includes administering an inhibitor to a subject.

[0041] In some examples, a method for modulating the immune system in a subject includes administering an effective amount of a serotonin receptor 5-HT 2A A method is provided that includes administering an inhibitor to a subject.

[0042] As used herein, the immune system encompasses all types of cells, compounds, compositions, and / or proteins that are part of the body's defense mechanisms against foreign substances and / or abnormal cells. In some examples, the immune system includes the innate immune system, such as NK cells, neutrophils, eosinophils, basophils, and monocytes. In some examples, the immune system includes the adaptive immune system, such as T lymphocytes, B lymphocytes, NK T cells, antigen-presenting cells, and dendritic cells. In some examples, the methods disclosed herein modulate CD8 T cells in a subject.

[0043] In some examples, the methods disclosed herein remove the suppression of immune cells by hepatocytes or liver-resident cells. In some examples, liver-resident cells may include, but are not limited to, non-parenchymal liver cells. In some examples, non-parenchymal liver cells include hepatic sinusoidal endothelial cells, hepatic stellate cells, etc. In some examples, liver-resident cells may include, but are not limited to, liver-resident immune cells. In some examples, liver-resident immune cells include liver-resident T cells, liver-resident NK cells, etc. In some examples, the methods disclosed herein remove the suppression of T lymphocytes. In some examples, the methods disclosed herein remove the suppression of CD8 T lymphocytes. In some examples, the methods disclosed herein remove the suppression of serotonin receptor 5-HT 2A The inhibitor improves suppression of the immune system, and optionally improves suppression of CD8 T cell function.

[0044] In some instances, the serotonin receptor 5-HT 2A The inhibitors act as adjuvant therapy / immunotherapy for subjects in need thereof.

[0045] In some instances, the serotonin receptor 5-HT 2A Inhibitors of serotonin receptor 5-HT enhance CD8 T cell activation. 2A The inhibitor increases the secretion of the pro-inflammatory cytokine IFNγ upon CD8 T cell activation.

[0046] Without wishing to be bound by theory, it is believed that the serotonin receptor 5-HT 2A Inhibitors (such as ketanserin) are believed to block immunosuppressive interactions between hepatocytes and tumor-specific immune cells. In some instances, the inventors have demonstrated that serotonin receptor 5-HT 2A We believe that inhibitors (such as ketanserin) surprisingly ameliorate hepatic nonparenchymal cell (NPC) suppression of CD8 T cell effector function. 2AWe have surprisingly found that treatment with inhibitors results in significant expansion and / or activation of CD8 T cell effector (function), and in some instances, serotonin receptor 5-HT 2A Inhibitors enhance the effectiveness of immune cells in immunotherapy. For example, serotonin receptor 5-HT 2A Inhibitors enhance the effectiveness of T cell-targeted immunotherapy.

[0047] In some examples, the functional phenotype of activated immune cells (such as activated CD8 T cells) can include, but are not limited to, upregulated CD25 expression, increased expression / production of IFNγ and / or TNFα, increased CD44 expression, increased expression / production of granzyme B, increased expression / production of perforin, and increased cytotoxicity (demonstrated by in vitro killing of target cells).

[0048] and the serotonin receptor 5-HT to enhance CD8 T cell activation. 2A Uses of inhibitors are also disclosed, one embodiment of which is the reduction of CD8 T cell suppression of non-parenchymal liver cells (NPCs). In some examples, a particular aspect of CD8 T cell activation that is enhanced is the secretion of the pro-inflammatory cytokine IFNγ.

[0049] Without wishing to be bound by theory, 5-HT 2A Targeting serotonin receptors and inhibiting hepatic non-parenchymal cells and CD8 T cell function enhances CD8 T cell activation. In some instances, this inhibition is due to 5-HT 2A It is mediated by disruption of the gene encoding (Htr2a in mice and HTR2A in humans).

[0050] In some instances, 5-HT 2A The inhibitor is 5-HT 2A It is a selective antagonist of

[0051] In some instances, 5-HT 2AInhibitors inhibit the serotonin receptor 5-HT by blocking its function in enhancing CD8 T cell activation 2A Agents targeting the 5-HT and / or serotonin receptors 2A and thus inhibit its function in suppressing CD8 T cell activation.

[0052] In some instances, 5-HT 2A The inhibitor is a compound having the formula: [ka] and pharmaceutically acceptable acid addition salts thereof and possible stereoisomers thereof, wherein R is hydrogen or C 1~6 is alkyl, Alk is C 1~4 is an alkanediyl, Q is a group of the following formula [ka] (In the formula, Y 1 and Y 2 are each independently O or S, R 2 is hydrogen, halo, C 1~6 Alkyl or C 1~6 alkyloxy, R 3 is hydrogen or halo) or Q is a group of the following formula [ka] (In the formula, R 4 is hydrogen or C 1~6 is alkyl, Z is -S-, -CH2- or -CR 5 =CR 6 wherein R 5 and R 6 are each independently hydrogen or C 1~6alkyl, and A is a divalent group -CH2-CH2-, -CH2-CH2-CH2- or -CR 7 =CR 8 and R 7 and R 8 are each independently hydrogen, halo, amino or C 1~6 alkyl), R 1 is the base of the following formula -X-Ar (c) wherein Ar is phenyl or substituted phenyl, said substituted phenyl having an amino group and / or 1, 2 or 3 halo atoms; X is >C=O, >CH-OH, >CH-OC(O)-R 9 , >CH2, >C(OC 1~6 alkyl)2, [ka] , >C=N-OH or >C=N-NH2; R 9 is hydrogen or C 1~6 is alkyl, wherein q is an integer of 2 or 3. or R 1 is the base of the following formula [ka] (In the formula, R 10 is hydrogen or C 1~6 alkyl, and R 11 , R 12 and R 13 are each independently hydrogen or halo. or R 1 is the base of the following formula [ka] (Wherein A is O or S, R 14 and R 15 are each independently hydrogen, halo, hydroxy, C 1~6Alkyloxy or C 1~6 alkyl).

[0053] In some examples, compounds of formula (I) according to the present disclosure may comprise R 1 is a group of formula (c).

[0054] In some examples, compounds of formula (I) according to the present disclosure include those wherein Q is a group of formula (a) and Y 1 and Y 2 are both oxygen atoms, and R 2 and R 3 are both hydrogen or Q is a group of formula (b) and R 4 is methyl and Z is -CR 5 =CR 6 - and R 5 and R 6 are independently hydrogen or methyl, and A is -CR 7 =CR 8 - and R 7 and R 8 are independently hydrogen or methyl, and / or in group (c), X is >C=O and Ar is halo-substituted phenyl.

[0055] In some examples, the compound according to the present disclosure is selected from the group consisting of 3-[2-[4-(4-fluorobenzoyl)-1-piperidinyl]ethyl]-2,4(1H,3H-quinazolinedione) (this compound is collectively designated ketanserin) and 3-[2-[4-(4-fluorobenzoyl)-1-piperidinyl]ethyl]-2,7-dimethyl-4H-pyrido-[1,2-a]-pyrimidin-4-one, and pharmaceutically acceptable acid addition salts thereof.

[0056] The compound of formula (I) can be used as it is or in the form of its acid addition salt.The acid addition salt can be conveniently obtained by treating the base form with a suitable acid, for example, inorganic acid, such as hydrohalic acid, for example, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or organic acid, for example, acetic acid, propanoic acid, hydroxyacetic acid, 2-hydroxypropanoic acid, 2-oxopropanoic acid, ethanedioic acid, propanedioic acid, butanedioic acid, (Z)-2-butenedioic acid, (E)-2-butenedioic acid, 2-hydroxybutanedioic acid, 2,3-dihydroxybutanedioic acid, 2-hydroxy-1,2,3-propanetricarboxylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, 4-methylbenzenesulfonic acid, cyclohexanesulfamic acid, 2-hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid, etc.

[0057] In some instances, 5-HT 2A The inhibitor is a compound having the formula: [ka] [In the formula, Z is [ka] R is hydrogen, a cyclic or linear or branched acyclic organyl group, a lower hydroxyalkyl group, a lower aminoalkyl group, or an aralkyl or heteroaralkyl group; n is 0, 1, or 2; X1 is methylene, vinylene, or an NH or N(lower alkyl) group; X2 is methylene, or when X1 is methylene or vinylene, X2 is methylene or a bond, or when X1 is methylene, X2 is O, S, NH, or N(lower alkyl) or a bond; Y1 is methylene and Y2 is methylene, vinylene, ethylene, propylene, or a bond; or Y1 is a bond and Y2 is vinylene, or Y1 is ethylene and Y2 is O, S, NH, or N(lower alkyl); Ar1 and Ar2 are independently an unsubstituted or substituted aryl or heteroaryl group; W is oxygen or sulfur; or It is a pharmaceutically acceptable salt, ester, or prodrug thereof.

[0058] Also provided is a pharmaceutical composition comprising an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, ester, or prodrug thereof.

[0059] In some instances, 5-HT 2A The inhibitor is a compound having the formula: [ka] wherein one of X and Y is CH2 and the other is selected from the group consisting of CH2, O, and S; The dotted line emanating from Z indicates an optional bond; when no bond is shown, Z is N, CH or COH; when a bond is shown, Z is C; Ar is phenyl, 2-thienyl, 3-thienyl, 2-furanyl, 3-furanyl, 2-pyrimidyl, 1-indolyl, 2-indolyl, 3-indolyl, 1-indol-2-onyl, 3-indol-2-onyl, 2- or 3-benzofuranyl, 2- or 3-benzothiophenyl, 1-naphthyl or 2-naphthyl, each optionally being halogen, lower alkyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, trifluoromethylsulfonyloxy, cycloalkyl, cycloalkyl-lower alkyl, nitro, amino, lower alkylamino, di-lower alkylamino, acylamino or C 1~2 substituted with alkylenedioxy; R 1is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, cycloalk(en)yl, cycloalk(en)yl-lower alk(en / yn)yl, aryl, aryl-lower alkyl, acyl, thioacyl, lower alkylsulfonyl, trifluoromethylsulfonyl, arylsulfonyl, R 1 is R 9 VCO- group (wherein V is O or S and R 9 is lower alkyl, cycloalkyl, cycloalkyl lower alkyl, or aryl), or R 1 is R 10 R 11 NCO-group or R 10 R 11 NCS- group (wherein R 10 and R 11 are independently hydrogen, lower alkyl, cycloalkyl, cycloalkyl-lower alkyl, or aryl, or R 10 and R 11 together with the N atom to which they are attached form a pyrrolidinyl, piperidinyl or perhydroazepine group, R 2 is hydrogen, lower alkyl, cycloalkyl, or cycloalkyl-lower alkyl; or R 1 and R 2 together with the N atom to which they are attached. [ka] wherein Q is C=O, C=S or CH, T is NH, S, O or CH, and m is 1 to 4 inclusive; R 3 ~R 5 are independently hydrogen, halogen, lower alkyl, lower alkylcarbonyl, phenylcarbonyl, halogen-substituted phenylcarbonyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, cycloalkyl, cycloalkyl-lower alkyl, or nitro; R6 and R 7 are each hydrogen or lower alkyl, or they are joined together to form a 3- to 7-membered carbocyclic ring; R 8 is hydrogen or lower alkyl, any alkyl, cycloalkyl or cycloalkylalkyl groups present are optionally substituted with one or two hydroxy groups, which in turn are optionally esterified with an aliphatic or aromatic carboxylic acid, and any aryl substituents present are optionally substituted with halogen, lower alkyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, trifluoromethylsulfonyloxy, cycloalkyl, cycloalkyl-lower alkyl or nitro, and pharmaceutically acceptable acid addition salts thereof.

[0060] In some instances, 5-HT 2A The inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate), 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound.

[0061] As used herein, the term pimavanserin refers to a member of the class of ureas in which three of the four hydrogens are replaced by a 4-fluorobenzyl group, a 1-methylpiperidin-4-yl group, and a 4-(isopropyloxy)benzyl group. Pimavanserin has CAS numbers 706779-91-1 or 706782-28-7, or is 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea, or the following formula: [ka] Also known as

[0062] As used herein, the term vorinancerin refers to a selective 5-HT 2A Vorinancerin has the CAS number 139290-65-6, or (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol, or the formula: [ka] Also known as

[0063] As used herein, the term ketanserin refers to a selective 5-HT 2A Ketanserin refers to a quinazoline derivative that acts as a serotonin antagonist. Ketanserin has the CAS number 74050-98-9 or 83846-83-7, or is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione], or the formula: [ka] Also known as

[0064] In some examples, the ketanserin described herein has CAS: 83846-83-7, or is 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione, or the following formula: [ka] It may also be ketanserin tartrate, also known as ketanserin tartrate.

[0065] In some instances, 5-HT may be based on the molecular structure of ketanserin. 2A Specific inhibitors of 5-HT may show efficacy in enhancing CD8 T cell activation. 2A The inhibitor of is ketanserin.

[0066] Pharmaceutically acceptable salts that can be mentioned include acid addition salts and base addition salts.Such salts can be formed by conventional means, for example, by reacting the compound of formula I in free acid or free base form with one or more equivalents of suitable acid or base, optionally in solvent or in the medium in which salt is insoluble, and then using standard techniques (for example, in vacuum, by lyophilization or by filtration) to remove said solvent or said medium.Salts can also be prepared by exchanging the counterion of the serotonergic compound in salt form with another counterion, for example, by using suitable ion exchange resin.

[0067] Examples of pharmaceutically acceptable salts include acid addition salts derived from mineral and organic acids, and salts derived from metals such as sodium, magnesium, or preferably potassium and calcium.

[0068] Examples of acid addition salts include acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, arylsulfonic acids (e.g., benzenesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, and p-toluenesulfonic acid), ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzoic acid, 4-acetamidobenzoic acid, butanoic acid, (+)-camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid (e.g., D-gluconic acid), glucuronic acid (e.g., 2-hydroxyethanesulfonic acid), methylsulf ... , D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-oxoglutaric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, isethionic acid, lactic acid (e.g., (+)-L-lactic acid and (±)-DL-lactic acid), lactobionic acid, maleic acid, malic acid (e.g., (-)-L-malic acid), malonic acid, (±)-DL-mandelic acid, metaphosphoric acid, methanesulfonic acid, 1 and acid addition salts formed with -hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, tartaric acid (e.g., (+)-L-tartaric acid), thiocyanic acid, undecylenic acid, and valeric acid.

[0069] Specific examples of salts are salts derived from mineral acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, metaphosphoric acid, nitric acid and sulfuric acid, salts derived from organic acids such as tartaric acid, acetic acid, citric acid, malic acid, lactic acid, fumaric acid, benzoic acid, glycolic acid, gluconic acid, succinic acid, arylsulfonic acids, and salts derived from metals such as sodium, magnesium, or preferably potassium and calcium.

[0070] As mentioned above, any solvates of the compounds and their salts are also included.Preferred solvates are those formed by incorporating a non-toxic pharmaceutically acceptable solvent (hereinafter referred to as a solvating solvent) into the solid structure (e.g., crystalline structure) of the compounds of the present invention.Examples of such solvents include water, alcohols (such as ethanol, isopropanol, and butanol) and dimethyl sulfoxide.Solvates can be prepared by recrystallizing the compounds of the present invention with a solvent or solvent mixture containing a solvating solvent.Whether a solvate is formed in any given case can be determined by analyzing the crystal of the compound using well-known standard techniques such as thermogravimetric analysis (TGE), differential scanning calorimetry (DSC), and X-ray crystallography.

[0071] The solvates may be stoichiometric or non-stoichiometric solvates. Particularly preferred solvates are hydrates, examples of which include hemihydrates, monohydrates and dihydrates.

[0072] The compounds of the present disclosure are generally administered as pharmaceutical preparations mixed with pharmaceutically acceptable adjuvants, diluents or carriers, which can be selected taking into consideration the intended route of administration and standard pharmaceutical practice.Such pharmaceutically acceptable carriers can be chemically inert to the active compounds and have no adverse side effects or toxicity under the conditions of use.Suitable pharmaceutical preparations are known in the art.For parenteral administration, parenterally acceptable aqueous solutions can be used that are pyrogen-free and have the required pH, isotonicity and stability.Suitable solutions are well known to those skilled in the art, and many methods are described in the literature.

[0073] Otherwise, the preparation of suitable formulations may be routinely accomplished by those skilled in the art using routine techniques and / or in accordance with standard and / or generally accepted pharmaceutical practice.

[0074] The amount of compound in any pharmaceutical formulation used in accordance with the present disclosure will depend on various factors, such as the severity of the condition being treated, the particular patient / subject being treated, and the compound(s) used. In any case, the amount of compound in the formulation can be determined routinely by one of ordinary skill in the art.

[0075] In some instances, the proliferative disorder is a tumor and / or cancer.

[0076] In some examples, the tumor / cancer is liver cancer (such as hepatocellular carcinoma), bone cancer, pancreatic cancer (such as pancreatic ductal adenocarcinoma (PDAC), pancreatic adenocarcinoma), skin cancer (such as melanoma, including but not limited to cutaneous or intraocular malignant melanoma), head and neck cancer, breast cancer, lung cancer, kidney cancer, uterine cancer, colon cancer, ovarian cancer, colorectal cancer, colon cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, cancer of the fallopian tubes, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, cancer of the endocrine system, thyroid cancer, parathyroid cancer, cancer of the adrenal gland, soft tissue sarcoma, urethral cancer, penile cancer, childhood solid tumor, lymphocytic lymphoma, bladder cancer, cancer of the kidney or ureter, renal pelvis cancer, central nervous system (CNS) neoplasm, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma. , pituitary adenoma, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, environmentally induced cancers including those induced by asbestos, hematological malignancies (including, for example, multiple myeloma), B-cell lymphoma, Hodgkin's lymphoma / primary mediastinal large B-cell lymphoma, non-Hodgkin's lymphoma, acute myeloid lymphoma, chronic myeloid leukemia, chronic lymphocytic leukemia, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, immunoblastic large cell lymphoma, precursor B-cell lymphoblastic lymphoma, mantle cell lymphoma, acute lymphocytic leukemia, mycosis fungoides, anaplastic large cell lymphoma, T-cell lymphoma, and precursor T-cell lymphoblastic lymphoma, and any combination of these cancers.

[0077] In some instances, the proliferative disease is liver cancer.

[0078] In some instances, the proliferative disease is hepatocellular carcinoma.

[0079] In some examples, the present disclosure further includes additional / combined therapies, including, but not limited to, immunotherapy (such as immune checkpoint blockade / ICB therapy / adoptive immune cell transfer therapy such as CAR-T cell therapy), chemotherapy, ablation therapy (local ablation therapy), such as radiation therapy or surgery (e.g., resection), transplantation, etc.

[0080] In some examples, the present disclosure further includes immunotherapy, such as, but not limited to, immune checkpoint blockade (ICB) therapy, including administration of monoclonal antibodies, and optionally, the immunotherapy may include the use of one or more of anti-PD-L1 (such as atezolizumab or nivolumab), anti-VEGFA (such as bevacizumab), anti-CTLA4 (such as ipilimumab), or anti-PD-1 (such as nivolumab) plus anti-CTLA4 (such as ipilimumab), and / or combinations thereof.

[0081] Also, a therapeutically effective amount of the serotonin receptor 5-HT 2A Also disclosed are methods of treating HCC, including administering the inhibitor in conjunction with immune checkpoint blockade (ICB) therapy. In some examples, the immune checkpoint blockade therapy can take the form of a combination of anti-PD-L1 and anti-VEGFA mAbs.

[0082] In some instances, the methods disclosed herein result in higher survival rates, delayed tumor growth, and lower mortality rates compared to the use of ICB therapy alone. In one such embodiment, the ICB therapy can take the form of a combination of anti-PD-L1 and anti-VEGFA mAbs.

[0083] In some instances, the combination therapy inhibits the serotonin receptor 5-HT 2A The inhibitors may be administered to the subject simultaneously, sequentially, or separately.

[0084] In some instances, the inhibitor is to be administered intravenously, subcutaneously, orally, sublingually, or intraperitoneally.

[0085] Also, the serotonin receptor 5-HT 2A Compositions comprising the inhibitors and immune checkpoint blockade / ICB therapy are also disclosed.

[0086] In another embodiment, the serotonin receptor 5-HT 2A Pharmaceutical compositions comprising the inhibitors and immune checkpoint blockade / ICB therapy are provided.

[0087] In some examples, the serotonin receptor inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate), 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound.

[0088] Also, the serotonin receptor 5-HT 2A Combination therapies including inhibitors and immune checkpoint blockade / ICB therapy are also disclosed.

[0089] In yet another embodiment, the serotonin receptor 5-HT 2Aand combination therapy including immune checkpoint blockade / ICB therapy, wherein the serotonin receptor inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate). , 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound.

[0090] In some examples, patients or subjects receiving treatment / therapy may show a reduction in the size or rate of tumor growth after administration of the inhibitors and / or combination therapies described herein. In some examples, this reduction may be observed at 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 weeks. In some examples, the reduction may be about 10%, about 20%, about 30%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95% or more. For example, at about week 12 following weekly administration of the inhibitors and / or combination therapies described herein, patients may exhibit about a 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more reduction in tumor growth (size and / or rate of growth).

[0091] Also disclosed are combination therapies described herein.

[0092] Also disclosed are the therapies / compositions / methods described herein.

[0093] Exemplary embodiments of the present disclosure will be better understood and readily apparent to those skilled in the art from the following discussion (and, where applicable, in conjunction with the drawings). It should be understood that other modifications to the serotonin inhibitors may be made without departing from the scope of the present invention. The exemplary embodiments are not necessarily mutually exclusive, as some may be combined with one or more embodiments to form new exemplary embodiments. The exemplary embodiments should not be construed as limiting the scope of the present disclosure. [Brief explanation of the drawings]

[0094] [Figure 1] In vitro screening of candidate small molecules that enhance hepatic NPC-activated CD8 T cell effector function is shown. Ovalbumin-specific OT-I CD8 T cells were co-cultured in a 96-well plate with an equal number of hepatic NPCs pulsed with ovalbumin peptide. Individual compounds from the Prestwick compound library of FDA-approved drugs were diluted into separate wells of the co-culture to a final concentration of 10 μM in each well. As a negative control, an equal volume of DMSO (to a final concentration of 0.5% v / v) was added to the co-culture (black symbols). As a positive control, OT-I CD8 T cells were activated with plate-bound αCD3ε (3 μg / mL) plus soluble αCD28 antibody (1 μg / mL). The functional phenotype of activated CD8 T cells was analyzed 4 days after activation using high-throughput multiparameter flow cytometry. Perturbations of each agent on each parameter were assessed by the direction and extent of perturbation from the mean of the negative control for each parameter and scored by the total number of positive perturbations. (A) Distribution of total positive perturbation scores across all 640 drugs screened. (B-F) Distribution of the magnitude and direction of perturbation (normalized by standard deviation from the mean of the negative controls) of the five functional parameters analyzed. Left panel—negative control, middle panel—50 preliminary hits, right panel—positive control. Each drug was tested in a single well and evaluated against 80 negative control wells and 80 positive control wells. [Figure 2]Figure 1 shows the effect of increasing concentrations of ketanserin on markers of CD8 T cell effector function during CD8 T cell activation by hepatic NPCs. Ketanserin at the indicated concentrations was added to the hepatic NPC-CD8 T cell cocultures in Figure 1. On day 4 post-activation, CD8 T cells were analyzed for effector function by flow cytometry for the indicated parameters. Statistical analysis was performed using the Kruskal-Wallis test and Dunn's multiple comparison test; *p<0.05, **p<0.01. Data represent two independent biological repeats. [Figure 3] The effects of genetic and / or pharmacological ablation of 5-HT2A activity on CD8 T cells activated by hepatic NPCs are shown. OT-I Cas9+ / + CD8 T cells were transduced with lentiviral vectors carrying gRNA sequences targeting either Htr2a or LacZ for CRISPR gene editing. (A) Immunoblot of 5-HT2A to assess the gene editing efficiency of each candidate gRNA. The red arrow (gRNA SEQ ID NO: 5) indicates the sequence used to create a CRISPR knockout of Htr2a in subsequent experiments. (B) Htr2a-KO or control gRNA (LacZ)-transduced OT-I cells were cocultured with ovalbumin-pulsed hepatic NPCs (Figure 1) in the presence of 5 μM ketanserin or an equivalent concentration of DMSO vehicle. On day 4 after activation, CD8 T cells were analyzed for effector function by flow cytometry for the indicated parameters. Statistical analysis using two-way ANOVA and Tukey's multiple comparison test, *p<0.05, **p<0.01, ***p<0.001. [Figure 4]Figure 1 shows that 5-HT2A is upregulated in CD8 T cells after activation in mice and humans. Polyclonal CD8 T cells isolated from wild-type mouse and healthy human PBMCs were activated in vitro for 4 days with plate-bound α-CD3 and soluble α-CD28 antibodies. (A) Immunoblot analysis of 5-HT2A and GAPDH in whole-cell lysates prepared from mouse CD8 T cells sampled at the indicated time points. Each lane was loaded with lysates equivalent to 5 x 10 cells. (B) Quantification of 5-HT2A protein levels relative to GAPDH loading control for samples in (A). (C) Immunoblot analysis of 5-HT2A and histone H3 in whole-cell lysates prepared from human CD8 T cells sampled at the indicated time points. Each lane was loaded with lysates equivalent to 5 x 10 cells. (D) Quantification of 5-HT2A protein levels relative to histone H3 loading control for samples in (C). (E) Kinetics of 5-HT2A protein expression during activation of CD8 T cells isolated from mice and two healthy human donors relative to loading control. [Figure 5] These results demonstrate that 5-HT2A inhibition using ketanserin during T cell activation enhances the cytotoxic effector phenotype in both mouse and human CD8 T cells. Polyclonal CD8 T cells isolated from (A) wild-type mice and (B) healthy human PBMCs were activated in vitro for 4 days with plate-bound α-CD3 and soluble α-CD28 antibodies. On day 4 post-activation, CD8 T cells were analyzed for effector function by flow cytometry for the indicated parameters. Statistical analysis was performed using a two-tailed Welch t-test; *p<0.05, **p<0.01. [Figure 6]Figure 1 shows the effect of 5-HT2A targeting using various methods in a spontaneously induced HCC mouse model. HCC was induced in immunocompetent mice by hydrodynamic injection of a plasmid expressing an oncogene and a firefly luciferase reporter. (a) Survival rates of HCC-bearing mice treated with 2.5 mg / kg ketanserin (red symbols) or an equal volume of 2.5% (v / v) DMSO vehicle (blue symbols) by intraperitoneal injection twice weekly, starting 25 days after tumor induction. Data are pooled from two independent replicates. Statistics were performed using the Mantel-Cox log-rank test. (b) Survival rates of mice induced with Htr2a-knockout (+Htr2a gRNA) or Htr2a-wild-type (+LacZ gRNA) HCC tumors. Data are pooled from two independent replicates. Statistical analysis was performed using the Mantel-Cox log-rank test. (c) Log2 fold change in tumor burden in HCC-bearing mice adoptively transferred with either Htr2a-KO or -WT tumor-specific CD8 T cells. Statistical analysis using the Mann-Whitney test.

[0095] Experimental data The present inventors identified the selective serotonin receptor antagonist ketanserin as a lead hit in an in vitro screen of the Prestwick library of US FDA-approved drugs for small molecules capable of improving hepatic nonparenchymal cell (NPC) suppression of CD8 T cell effector function (Figure 1). Ketanserin achieved a high score (7 out of a maximum possible 9) on a number of positive perturbation parameters of CD8 T cells (Figure 1A). Ketanserin treatment of hepatic NPC-activated CD8 T cells increased their survival and proliferation rates (Figure 1B) and also resulted in increased expression of the activation marker CD25 (Figure 1C) and the effector cytokines IFNγ and TNFα (Figures 1D and 1E), but not the cytotoxic effector molecule granzyme B (Figure 1F).

[0096] We subsequently validated this hit molecule by incubating CD8 T cells activated in vitro with hepatic NPCs with increasing doses of ketanserin tartrate and found that treatment with ketanserin had a dose-dependent effect on the number of viable cells, CD25 expression, and INFγ and TNFα secretion (Figure 2). As further validation, we disrupted the Htr2a locus (5-HT 2A We generated CRISPR-Cas9-edited CD8 T cells (Fig. 3A) encoding Htr2a, and activated these Htr2a-KO cells with hepatic NPCs in the presence or absence of ketanserin. We found that genetic disruption of Htr2a significantly reduced the uptake of 5-HT. 2A We found that inhibition of IL-16 by ketanserin recapitulated the phenotype of increased proliferation, CD25 expression, and INFγ secretion that we previously observed (FIG. 3B).

[0097] To determine whether 5-HT2A serotonin receptors are expressed by human CD8 T cells, we examined the expression of 5-HT2A receptors during activation of mouse and human CD8 T cells by immunoblot analysis. 2A The kinetics of expression was profiled (Figure 4). 2A We found that expression of ketanserin was consistently increased after in vitro activation of both mouse (consistent with data from Leon-Ponte et al. (Blood, 2007)) and human CD8 T cells (a novel finding not previously described in the literature). These data indicate that the molecular target of the tool compound ketanserin is indeed present in activated CD8 T cells and can therefore be inhibited by the specific action of ketanserin.

[0098] We have used ketanserin to inhibit 5-HT in mouse and human CD8 T cells during activation. 2A The effects of ketanserin on inhibiting granzyme B activity after in vitro activation were further compared (Figure 5). In both mouse and human CD8 T cells, treatment with ketanserin significantly inhibited granzyme B activity after in vitro activation. +In contrast to mouse CD8 T cells, human CD8 T cells showed a trend toward increased cell number and INFγ expression, but these were not significant.

[0099] We evaluated the therapeutic effects of ketanserin administration in our custom MITCH model of HCC (SIGN / TDF / 121). We previously described that the MITCH model of HCC only partially responds to combined monoclonal antibody blockade of PD-L1 and VEGFR2, mimicking the resistance observed when HCC patients were treated with the combination of atezolizumab (anti-PD-L1) and bevacizumab (anti-VEGFA), which is approved as a first-line HCC therapy in the clinic. Mice treated with ketanserin survived significantly longer than mice in the control cohort, and tumor growth and mortality were generally delayed (Figure 6A). Because mice bearing Htr2a-deficient HCC tumors showed no significant difference in survival or spontaneous tumor rejection compared to mice bearing Htr2a wild-type HCC tumors, the protective effect of ketanserin is likely mediated by actions on cells other than HCC tumor cells (Figure 6B). Furthermore, adoptive transfer of HCC-bearing mice with Htr2a-KO tumor-specific CD8 T cells also more potently reduced tumor burden compared with mice treated with the same number of Htr2a-wild-type CD8 T cells (Figure 6C).

[0100] Thus, these data suggest that the serotonin receptor 5-HT 2A This study provides mounting evidence that inhibition of T cell-targeted immunotherapy of HCC could potentially be an effective adjuvant for T cell-targeted immunotherapy.

[0101] Application Examples The serotonin receptor 5-HT 2A Embodiments of inhibitors of provide an alternative adjuvant immunotherapy for treating proliferative diseases.

[0102] Advantageously, 5-HT 2ASpecifically targeting serotonin receptors results in the specific activation of tumor-specific CD8 T cells by improving the suppression of CD8 T cells by nonparenchymal liver cells.

[0103] Those skilled in the art will appreciate that other changes and / or modifications can be made to the embodiments disclosed herein without departing from the spirit or scope of the present disclosure as broadly described. For example, in the description herein, features of different exemplary embodiments can be mixed, combined, interchanged, incorporated, adopted, modified, encompassed, etc. across the different exemplary embodiments. The present embodiments are therefore to be considered in all respects as illustrative and non-restrictive.

Claims

1. Serotonin receptor 5-HT for use in treating a proliferative disorder in a subject in need thereof - Patent application ２Ａ Inhibitors or serotonin receptor 5-HT ２Ａ A composition comprising an inhibitor.

2. Serotonin receptor 5-HT in the manufacture of a medicament for treating a proliferative disorder in a subject in need thereof - Patent Application 20070122997 ２Ａ Use of inhibitors.

3. A method of treating a proliferative disorder in a subject in need thereof, comprising administering an effective amount of a serotonin receptor 5-HT antagonist to a subject in need thereof. ２Ａ administering an inhibitor to said subject.

4. The serotonin receptor 5-HT ２Ａ 5-HT serotonin receptor inhibitors for use according to claim 1, wherein the inhibitor improves the suppression of the immune system and optionally improves the suppression of CD8 T cell function. ２Ａ Inhibitor or or use according to claim 2 or method according to claim 3.

5. The 5-HT ２Ａ The inhibitor is 5-HT ２Ａ 5. The method of claim 1, wherein the serotonin receptor 5-HT2 receptor is a selective antagonist of the serotonin receptor 5-HT2 receptor. ２Ａ Inhibitor or the use according to claim 2 or 4 or the method according to claim 3 or 4.

6. The 5-HT ２Ａ Inhibitors inhibit the serotonin receptor 5-HT by inhibiting its function in enhancing CD8 T cell activation. ２Ａ and / or the serotonin receptor 5-HT ２Ａ 5. The method of claim 1, wherein the serotonin receptor 5-HT2 receptor is an agent that targets the overall function of the serotonin receptor 5-HT2 receptor and thus inhibits its function in enhancing CD8 T cell activation. ２Ａ Inhibitor or use according to any one of claims 2 or 4-5 or method according to claim 3 or any one of claims 4-5.

7. The 5-HT ２Ａ The inhibitor is a compound having the formula: 【Chemistry 1】 and pharmaceutically acceptable acid addition salts thereof and possible stereoisomers thereof, wherein R is hydrogen or C １～６ is alkyl, Alk is C １～４ is an alkanediyl, Q is a group of the following formula: 【Chemistry 2】 (In the formula, Y １ and Y ２ are each independently O or S; R ２ is hydrogen, halo, C １～６ Alkyl or C １～６ alkyloxy, R ３ is hydrogen or halo; or Q is a group of the following formula: 【Transformation 3】 (In the formula, R ４ is hydrogen or C １～６ is alkyl, Z is -S-, -CH ２ -or-CR ５ =CR ６ -, and said R ５ and R ６ are each independently hydrogen or C １～６ alkyl and A is a divalent group -CH ２ -CH ２ -, -CH ２ -CH ２ -CH ２ -or-CR ７ =CR ８ -, and said R ７ and R ８ are each independently hydrogen, halo, amino or C １～６ alkyl), R １ is the basis of the following formula -X-Ar (c) wherein Ar is phenyl or substituted phenyl, said substituted phenyl having an amino group and / or 1, 2 or 3 halo atoms; X is >C=O, >CH-OH, >CH-O-C(O)-R ９ , >CH ２ , >C(OC １～６ alkyl) ２ , 【Chemistry 4】 , >C=N-OH or >C=N-NH ２ and The R ９ is hydrogen or C １～６ is alkyl, wherein q is an integer of 2 or 3. That is, or R １ is the basis of the following formula 【Transformation 5】 (In the formula, R １０ is hydrogen or C １～６ alkyl, and R １１ , R １２ and R １３ are each independently hydrogen or halo; or R １ is the basis of the following formula 【Transformation 6】 (Wherein A is O or S, R １４ and R １５ are each independently hydrogen, halo, hydroxy, C １～６ Alkyloxy or C １～６ alkyl), Serotonin receptor 5-HT for the use according to any one of claims 1 or 4 to 6 ２Ａ Inhibitor or the use according to claim 2 or any one of claims 4 to 6 or the method according to claim 3 or any one of claims 4 to 6.

8. The 5-HT ２Ａ The inhibitor is a compound having the formula: 【Transformation 7】 [In the formula, Z is 【Transformation 8】 R is hydrogen, a cyclic or linear or branched acyclic organyl group, a lower hydroxyalkyl group, a lower aminoalkyl group, or an aralkyl or heteroaralkyl group; n is 0, 1, or 2; X １ is methylene, vinylene, or an NH or N(lower alkyl) group; X ２ is methylene, or X １ is methylene or vinylene, X ２ is methylene or a bond, or X １ is methylene, X ２ is O, S, NH, or N(lower alkyl) or a bond; Y １ is methylene, and Y ２ is methylene, vinylene, ethylene, propylene, or a bond; Y １ is a bond, and Y ２ is vinylene, or Y １ is ethylene, and Y ２ is O, S, NH, or N(lower alkyl); Ar １ and Ar ２ are independently unsubstituted or substituted aryl or heteroaryl groups; W is oxygen or sulfur; or a pharmaceutically acceptable salt, ester, or prodrug thereof; Serotonin receptor 5-HT for the use according to claim 1 or any one of claims 4 to 7 ２Ａ Inhibitor or the use according to claim 2 or any one of claims 4 to 7 or the method according to claim 3 or any one of claims 4 to 7.

9. The 5-HT ２Ａ The inhibitor is a compound having the formula: 【Chemistry 9】 wherein one of X and Y is CH ２ and the other is CH ２ , O, and S; The dotted line emanating from Z indicates an optional bond; when no bond is shown, Z is N, CH or COH; when a bond is shown, Z is C; Ar is phenyl, 2-thienyl, 3-thienyl, 2-furanyl, 3-furanyl, 2-pyrimidyl, 1-indolyl, 2-indolyl, 3-indolyl, 1-indol-2-onyl, 3-indol-2-onyl, 2- or 3-benzofuranyl, 2- or 3-benzothiophenyl, 1-naphthyl or 2-naphthyl, each optionally being halogen, lower alkyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, trifluoromethylsulfonyloxy, cycloalkyl, cycloalkyl-lower alkyl, nitro, amino, lower alkylamino, di-lower alkylamino, acylamino or C １～２ substituted with alkylenedioxy; R １ is hydrogen, lower alkyl, lower alkenyl, lower alkynyl, cycloalk(en)yl, cycloalk(en)yl-lower alk(en / yn)yl, aryl, aryl-lower alkyl, acyl, thioacyl, lower alkylsulfonyl, trifluoromethylsulfonyl, arylsulfonyl, R １ is R ９ VCO- group (wherein V is O or S, and R ９ is lower alkyl, cycloalkyl, cycloalkyl lower alkyl, or aryl), or R １ is R １０ R １１ NCO-group or R １０ R １１ NCS- group (in the formula, R １０ and R １１ are independently hydrogen, lower alkyl, cycloalkyl, cycloalkyl-lower alkyl, or aryl, or R １０ and R １１ together with the N atom to which they are attached form a pyrrolidinyl, piperidinyl or perhydroazepine group, R ２ is hydrogen, lower alkyl, cycloalkyl, or cycloalkyl-lower alkyl; or R １ and R ２ together with the N atom to which they are attached form the group 【Chemistry 10】 (Wherein Q is C=O, C=S or CH ２ and T is NH, S, O or CH ２ and m is 1 to 4 inclusive; R ３ ~R ５ are independently hydrogen, halogen, lower alkyl, lower alkylcarbonyl, phenylcarbonyl, halogen-substituted phenylcarbonyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, cycloalkyl, cycloalkyl-lower alkyl, or nitro; R ６ and R ７ are each hydrogen or lower alkyl, or they are joined together to form a 3- to 7-membered carbocyclic ring; R ８ is hydrogen or lower alkyl, any alkyl, cycloalkyl or cycloalkylalkyl groups present are optionally substituted with one or two hydroxy groups, which in turn are optionally esterified with an aliphatic or aromatic carboxylic acid, and any aryl substituents present are optionally substituted with halogen, lower alkyl, lower alkoxy, lower alkylthio, hydroxy, lower alkylsulfonyl, cyano, trifluoromethyl, trifluoromethylsulfonyloxy, cycloalkyl, cycloalkyl-lower alkyl or nitro; and pharmaceutically acceptable acid addition salts thereof. Serotonin receptor 5-HT for the use according to claim 1 or any one of claims 4 to 8 ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 8 or method according to claim 3 or any one of claims 4 to 8.

10. The 5-HT ２Ａ The inhibitors include [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate), 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4 10. The serotonin receptor 5-HT1 receptor antagonist for use according to claim 1 or any one of claims 4 to 9, wherein the serotonin receptor 5-HT1 receptor antagonist is selected from the group consisting of (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 9 or method according to claim 3 or any one of claims 4 to 9.

11. Serotonin receptor 5-HT receptor agonist for use according to claim 1 or any one of claims 4 to 10, wherein the proliferative disease is a tumor and / or cancer. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 10 or method according to claim 3 or any one of claims 4 to 10.

12. The serotonin receptor 5-HT receptor inhibitor for use according to any one of claims 1 and 4 to 11, wherein the proliferative disease is liver cancer. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 11 or method according to claim 3 or any one of claims 4 to 11.

13. The serotonin receptor 5-HT receptor inhibitor for use according to any one of claims 1 and 4 to 12, wherein the proliferative disease is hepatocellular carcinoma. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 12 or method according to claim 3 or any one of claims 4 to 12.

14. The method of claim 1 or any one of claims 4 to 13, further comprising one or more combination therapies selected from the group consisting of immunotherapy, chemotherapy, ablation therapy, and transplantation. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 13 or method according to claim 3 or any one of claims 4 to 13.

15. 15. The method of claim 1 or any one of claims 4 to 14, further comprising immunotherapy such as, but not limited to, immune checkpoint blockade (ICB) therapy, including administration of monoclonal antibodies, optionally wherein said immunotherapy may comprise the use of one or more of anti-PD-L1 (such as atezolizumab or nivolumab), anti-VEGFA (such as bevacizumab), anti-CTLA4 (such as ipilimumab), or anti-PD-1 (such as nivolumab) plus anti-CTLA4 (such as ipilimumab), and / or combinations thereof. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 14 or method according to claim 3 or any one of claims 4 to 14.

16. The combination therapy is ２Ａ The serotonin receptor 5-HT inhibitor for use according to claim 1 or any one of claims 4 to 15 is to be administered to the subject simultaneously, sequentially or separately with the inhibitor. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 15 or method according to claim 3 or any one of claims 4 to 15.

17. 17. The method of claim 1 or any one of claims 4 to 16, wherein the inhibitor is to be administered intravenously, subcutaneously, orally, sublingually, or intraperitoneally. ２Ａ Inhibitor or use according to claim 2 or any one of claims 4 to 16 or method according to claim 3 or any one of claims 4 to 16.

18. Serotonin receptor 5-HT ２Ａ A pharmaceutical composition comprising an inhibitor and immune checkpoint blockade / ICB therapy.

19. 18. The pharmaceutical composition of claim 17, wherein the serotonin receptor inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate), 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound.

20. the serotonin receptor inhibitor is [3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione] (ketanserin), 2,3-dihydroxybutanedioic acid, 3-[2-[4-(4-fluorobenzoyl)piperidin-1-yl]ethyl]-1H-quinazoline-2,4-dione (ketanserin tartrate), 1-[(4-fluorophenyl)methyl]-1-(1-methylpiperidin-4-yl)-3-[[4-(2-methylpropoxy)phenyl]methyl]urea (pimavanserin), (R)-(2,3-dimethoxyphenyl)-[1-[2-(4-fluorophenyl)ethyl]piperidin-4-yl]methanol (borinancerin) or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutically functional derivative of such a compound; ２Ａ inhibitors and combination therapies including immune checkpoint blockade / ICB therapy.

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