Drug application of small molecule inhibitor targeting CD47

Abstract

The invention discloses a compound with a structure shown in a formula I or a pharmaceutically acceptable salt thereof, or a compound with a structure shown in a formula II or a pharmaceutically acceptable salt thereof, or a compound with a structure shown in a formula III or a pharmaceutically acceptable salt thereof. The invention discloses application of a compound as shown in a formula I or a pharmaceutically acceptable salt thereof, or a compound as shown in a formula II or a pharmaceutically acceptable salt thereof, or a compound as shown in a formula III or a pharmaceutically acceptable salt thereof in preparation of a CD47 inhibitor. The invention further discloses application of the compound with the structure shown in the formula I or pharmaceutically acceptable salt of the compound, or the compound with the structure shown in the formula II or pharmaceutically acceptable salt of the compound, or the compound with the structure shown in the formula III or pharmaceutically acceptable salt of the compound in preparation of drugs for treating malignant tumors caused by CD47 overexpression. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine and relates to a small molecule inhibitor targeting CD47 and its application in preparing a CD47 inhibitor and preparing a drug for treating malignant tumors caused by overexpression of CD47. Background Art

[0002] Currently, the incidence and mortality of hematological malignancies, including non-Hodgkin's lymphoma and acute myeloid leukemia, as well as solid tumors, such as colorectal cancer, ovarian cancer, breast cancer, bladder cancer, and head and neck cancer, are showing a significant upward trend worldwide. Tumor immunotherapy can block immune checkpoints through antibodies or small molecule inhibitors to restore the immune clearance of the above tumors by the immune system. Among them, macrophages play an important role in nonspecific tumor immunotherapy, and the macrophage immune checkpoint CD47 is an important cause of the loss of phagocytosis in the tumor microenvironment (TME).

[0003] The CD47 protein is a transmembrane protein and cell surface glycoprotein molecule that belongs to the immunoglobulin superfamily. CD47 on the surface of tumor cells inhibits the phagocytosis of macrophages by interacting with signal regulatory protein alpha (SIRPα) on the surface of macrophages. The mechanism is that after SIRPα interacts with CD47, it promotes the localization of SIRPα to the phagocytic synapse, activates Src homology region 2 domain phosphatase-1, and finally inhibits the accumulation of non-myosin IIA on the cell membrane, preventing tumor cells from being phagocytosed. The CD47-SIRPα axis is a key immune checkpoint in different cancers, including hematological malignancies. Preclinical and clinical studies have used a variety of CD47-targeted antibodies to treat tumors and have shown good efficacy, which provides convincing evidence for the therapeutic potential of targeting the CD47-SIRPα axis in tumors.

[0004] However, so far, no small molecule inhibitor targeting CD47 has entered the clinical trial stage on the market. As inhibitors that also act on the CD47 target, small molecule inhibitors can also effectively treat hematological malignancies such as non-Hodgkin's lymphoma and acute myeloid leukemia, as well as solid tumors such as colorectal cancer, ovarian cancer, breast cancer, bladder cancer and head and neck cancer, and inhibit tumor cell metastasis. In addition, compared with antibodies targeting CD47, small molecule inhibitors targeting CD47 have more advantages. First, small molecule inhibitors are not immunogenic, and compared with antibodies, they have a shorter half-life and are more targeted, which makes the side effects of small molecule inhibitors in the body more controllable during treatment. Secondly, CD47 is a ubiquitously expressed cell surface protein that is also expressed on red blood cells. Some CD47 antibodies bind to high-density determinant clusters on the red blood cell membrane and cause red blood cell agglutination, while specific small molecule inhibitors may avoid this. At the same time, small molecule inhibitors also have the potential for oral administration, which can improve patient compliance. Therefore, finding CD47 small molecule inhibitors is an urgent problem to be solved. Summary of the invention

[0005] The present invention aims to develop a small molecule inhibitor targeting CD47 with good activity and effect, and use it in the treatment of malignant tumors caused by overexpression of CD47.

[0006] The inventors constructed an inhibitor screening model targeting CD47. The model predicts the interaction between the target protein and the small molecule compound library, which greatly reduces the number of compounds screened in experiments, shortens the experimental cycle, reduces the cost of drug development, and improves the efficiency of active compound discovery. The inventors obtained candidate compounds with good predicted activity through virtual screening, and then verified the activity in vitro based on the CD47-SIRPα binding kit, and further screened small molecule inhibitors with good inhibitory effects on CD47 protein.

[0007] The object of the present invention is to provide a compound having a structure as shown in formula I (denoted as compound SWY-AK-309) or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in formula II (denoted as compound SWY-AM-598) or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in formula III (denoted as compound SWY-AF-747) or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug:

[0008]

[0009] The chemical name of compound SWY-AK-309: ethyl 2-[(4-isopropylbenzoyl)amino]-4-methyl-1,3-thiazole-5-carboxylate.

[0010] The chemical name of compound SWY-AM-598 is: 6-chloro-3,10-dimethoxybenzo[4,5]thieno[2,3-c]quinoline.

[0011] The chemical name of compound SWY-AF-747 is: 4-(1-benzofuran-2-ylcarbonyl)-5-[4-(benzyloxy)-3-methoxyphenyl]-3-hydroxy-1-(2-phenylethyl)-2,5-dihydro-1H-pyrrol-2-one.

[0012] The pharmaceutically acceptable salts of the compound of formula I are non-toxic anionic salts, including but not limited to chloride, bromide, iodide, sulfate, hydrochloride, maleate, fumarate, oxalate, lactate, tartrate, gluconate, methanesulfonate and 4-toluenesulfonate.

[0013] The pharmaceutically acceptable salts of the compound represented by Formula II are non-toxic anionic salts, including but not limited to chloride, bromide, iodide, sulfate, hydrochloride, maleate, fumarate, oxalate, lactate, tartrate, gluconate, methanesulfonate and 4-toluenesulfonate.

[0014] The pharmaceutically acceptable salt of the compound shown in Formula III is a non-toxic cationic salt, including but not limited to sodium salt, potassium salt, calcium salt, magnesium salt or a cationic salt formed by the compound shown in Formula III and choline, ethanolamine, diethanolamine, benzylethylamine, piperazine, N-methylglucosamine or tromethamine.

[0015] The half maximal inhibitory concentration (IC) of the three compounds of the present invention on CD47-SIRPα 50) are 3.57μM, 4.79μM and 12.473μM respectively. Compared with CD47 antibodies, the small molecule inhibitors targeting CD47 of the compounds of the present invention have the advantages of short half-life, controllable side effects, no immunogenicity, and avoidance of blood toxicity. The three compounds or their pharmaceutically acceptable salts, tautomers, prodrugs, and pharmaceutically acceptable salts of prodrugs described in the present invention can target CD47 protein, promote the phagocytosis of tumor cells by macrophages by blocking the CD47-SIRPα signaling pathway, thereby inhibiting tumor metastasis and achieving the purpose of treating tumors. The small molecule inhibitor targeting CD47 described in the present invention can target hematological malignancies such as non-Hodgkin's lymphoma and acute myeloid leukemia, as well as solid tumors such as colorectal cancer, ovarian cancer, breast cancer, bladder cancer, and head and neck cancer, and can also be extended to the treatment of other malignant tumors overexpressing CD47.

[0016] Another object of the present invention is to provide the use of a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula III or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug in the preparation of a CD47 inhibitor.

[0017] Another object of the present invention is to provide a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula III or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug in the preparation of a drug for treating malignant tumors caused by overexpression of CD47.

[0018] The malignant tumors caused by overexpression of CD47 are malignant tumors of the blood system, colorectal cancer, ovarian cancer, breast cancer, glioblastoma, bladder cancer, prostate cancer, and head and neck cancer.

[0019] The malignant tumors of the blood system are acute myeloid leukemia and chronic myeloid leukemia.

[0020] Another object of the present invention is to provide a pharmaceutical composition, which is a preparation prepared with a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula III or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug as an active ingredient and a pharmaceutically acceptable carrier.

[0021] The preparation is a solid dosage form, a liquid dosage form, a gas dosage form, or a semisolid dosage form.

[0022] The carrier comprises at least one of an excipient, a binder, a disintegrant, a lubricant, a flavoring agent, a fragrance, a colorant and a sweetener. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 IC of compounds SWY-AK-309, SWY-AM-598, and SWY-AF-747 against CD47-SIRPα 50 curve. DETAILED DESCRIPTION

[0024] The present invention mainly relates to the development process of CD47 small molecule inhibitors and the study of compound-protein interaction at the molecular level (based on the inhibition rate and IC of CD47-SIRPα binding kit). 50 In order to prove that the present invention inhibits the activity of CD47 protein, the following pharmacological experiments will be carried out.

[0025] Example 1

[0026] Reagents: CD47 / SIRP ALPHA BINDING KITSAS (Shanghai Medicilon Biotechnology Co., Ltd.). The detection kit contains frozen Tag1-CD47, Tag2-SIRP alpha, CD47 / SIRP alpha standards, Anti-Tag1Eu Cryptate reagent, Anti-Tag2 XL665 antibody and PPI Europium detection buffer.

[0027] Compounds to be tested: Compound SWY-AK-309, Compound SWY-AM-598, Compound SWY-AF-747.

[0028] Data processing method: The ratio was calculated according to Emission Ratio (ER) = Em665 / Em615, the ER in the compound well was recorded as ERcompound, the ER in the control well was recorded as ERvehicle, and the ER in the blank control well was recorded as ERblank (for specific operations, please refer to the kit operation flow https: / / www.cisbio.cn / human-cd47-sirp-alpha-biochemical-binding-kit-44631), and the inhibition rate of the test compound at different concentrations was calculated to illustrate the inhibitory ability of the compound on CD47-SIRPα binding. The inhibition rate was calculated using the following formula:

[0029] Inhibition rate = (ERvehicle-ERcompound) / (ERvehicle-ERblank)×100%

[0030] IC 50 Determination: The IC values ​​of the three compounds were determined using the activity detection method provided by the CD47 / SIRP ALPHA BINDING KITAS kit (https: / / www.cisbio.cn / human-cd47-sirp-alpha-biochemical-binding-kit-44631) and SpectraMax i3x Multi-Mode Reader (Molecular Devices). 50 ,IC 50 Curve Figure 1 .

[0031] IC of compound SWY-AK-309 against CD47-SIRPα 50 The IC value of compound SWY-AM-598 on CD47-SIRPα is 3.569 μM. 50 The IC value of compound SWY-AF-747 for CD47-SIRPα is 4.792 μM. 50 It was 12.473 μM, indicating that all three compounds had strong inhibitory ability on CD47-SIRPα binding.

Claims

1. A compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula III or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug:

2. Use of a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula III or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug in the preparation of a CD47 inhibitor.

3. Use of a compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula II or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in Formula III or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug in the preparation of a drug for treating malignant tumors caused by overexpression of CD47.

4. The use according to claim 3, characterized in that: The malignant tumors caused by overexpression of CD47 are malignant tumors of the blood system, colorectal cancer, ovarian cancer, breast cancer, glioblastoma, bladder cancer, prostate cancer, and head and neck cancer.

5. The use according to claim 4, characterized in that: The malignant tumors of the blood system are acute myeloid leukemia and chronic myeloid leukemia.

6. A pharmaceutical composition, characterized in that: The pharmaceutical composition is a preparation prepared with a compound having a structure as shown in formula I of claim 1 or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in formula II or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug, or a compound having a structure as shown in formula III or a pharmaceutically acceptable salt, tautomer, prodrug, or a pharmaceutically acceptable salt of a prodrug as an active ingredient and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition according to claim 6, characterized in that: The preparation is a solid dosage form, a liquid dosage form, a gas dosage form, or a semisolid dosage form.

8. The pharmaceutical composition according to claim 6, characterized in that: The carrier comprises at least one of an excipient, a binder, a disintegrant, a lubricant, a flavoring agent, a fragrance, a colorant and a sweetener.

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