Patent Application
20230391783
The present application relates to a field of organic synthesis and pharmaceutical chemistry, in particular to compounds containing 2,4-thiazole ring, method for preparing same and use thereof.
Any discussion of the prior art throughout the specification should not be taken as an admission that such prior art is widely known or forms part of the common general knowledge in the art.
Autoimmune disease is a disease in which T and B cells are over-activated in self-reaction, and cause damage to their own tissues and organs as a result of an immune response to their own antigens, such as systemic lupus erythematosus (SLE) and psoriasis. Epidemiological surveys show that there are millions of patients with SLE in China. At present, there are still no new chemical drugs for such autoimmune diseases worldwide. Clinical treatment is mainly based on the use of glucocorticoids in combination with non-specific anti-inflammatory and immunosuppressive drugs, which delays the progress of the disease to a certain extent, but long-term use will cause the decline of patients' immune function and cause a variety of complications.
Studies have proved that B lymphocytes and T lymphocytes play an important role in autoimmune diseases. It is an important topic in medical and pharmaceutical research to develop new drugs that can inhibit the proliferation and activation of immune cells and reduce the abnormal immune response of the body to treat autoimmune diseases.
Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor expressed mainly in the nucleus and can be activated by a range of compounds, such as the carcinogen 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), which has the property of promoting tumor growth and activating immune cells when activated by agonists such as TCDD. Studies have shown that AhR is a key transcription factor of T helper 22 (Th22) cell and plays an important role in immune response, thus AhR receptor has great potential in the treatment of autoimmune diseases.
In the absence of ligands, AhR is in the cytoplasm and exists as part of a protein complex consisting of heat shock protein (HSP) 90, p23, and AhR. Upon binding of a ligand such as TCDD, the AhR complex is activated and translocated to the nucleus, where AhR is released from chaperone protein and interacts with arylhydroarbon nuclear translocator (ARNT). Chaperone protein protects AhR from protein hydrolysis and retain a structure conducive to ligand binding. AhR-ARNT heterodimer is associated with signaling factors (e.g. chromatin remodeling factors, histone acetyltransferase and transcription factors), and ultimately binds to distal regulatory elements (DREs) or aryl hydrocarbon response elements (AHREs) to facilitate transcriptional regulation. Designing AhR-targeted immunomodulators is of great significance for the treatment of autoimmune diseases.
The present application provides compounds containing 2,4-thiazole ring in the structure thereof and pharmaceutically acceptable salts or isomers thereof, wherein the compounds have less toxic and side effects and have an inhibitory effect on the activity of immune cells.
In particular, the present application provides the following technical features, one or more of which constitute a technical solution for the present application.
In a first aspect of the present application, the present application provides a compound containing a 2,4-thiazole ring or a pharmaceutically acceptable salt or isomer thereof, the compound having a structure of Formula X:
And, in the compounds of Formula X2, R3 is not hydroxyl when R2 is alkyl.
Inventors have found that certain compounds tend to have severe toxicity and side effects while having an inhibitory effect on the activity of immune cells, such as in some embodiments where R2 is C1-C3 alkyl and R3 is hydrogen, and, in the case of the structure of Formula X2, where R2 is alkyl and R3 is hydroxyl. However, the compounds in the present application have no obvious toxicity at the test concentration, and have high safety index, good oral bioavailability and good developability.
In the present application, the term “C1-C6 alkyl” refers to a straight-chain, saturated hydrocarbon radical containing 1 to 6 carbon atoms, including, without limitation, methyl, ethyl, propyl, etc.
The term “C5-C15 alkenyl” refers to a straight-chain or branched hydrocarbon radical with one or more double bonds and containing 5 to 15 carbon atoms.
The term “3-10 membered heterocyclyl” refers to a saturated or partially saturated cyclic group having 3-10 ring atoms of which 1-3 are heteroatoms selected from the group consisting of nitrogen, oxygen and S(O)m (where m is an integer from 0 to 2), the remaining ring atoms being carbon atoms; for example, propylene oxide, tetrahydrofuranyl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc.
The term “C6-C12 aryl” refers to an aromatic ring group containing 6-10 ring atoms, but without heteroatoms in the ring atoms, such as phenyl, naphthyl, biphenyl, etc.
The term “5-12 membered heteroaryl” refers to an aromatic ring group of 5-12 ring atoms containing 1-4 ring heteroatoms. The heteroatom each independently selected from nitrogen, oxygen or sulfur. The heteroaryl may be a monocyclic heteroaryl having 5-7 ring atoms or a bicyclic heteroaryl having 7-12 ring atoms. It is sufficient that one of the bicyclic heteroaryl rings is a heteroaryl, and the other may be an aromatic or non-aromatic ring, with or without a heteroatom. In addition, bicyclic heteroaryl may be a fused ring structure in which two heterocycles share a common ring edge, or in which the two heterocycles are joined directly, such as by a single bond. Examples of heteroaryl groups include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, pyridyl, pyrimidyl, furyl, thienyl, isoxazolyl, indolyl, etc.
The term “sterol group” refers to a group of perhydrocyclopentanophenanthrene derivatives fused by three cyclohexane and one cyclopentane, such as sitosterol group, cholesterol group, ergosterol group, solasodine group and protodioscin group etc.
The term “3-10-membered cycloalkyl” means a group containing one or more saturated and/or partially saturated rings, all of which are carbon atoms, comprising from 3 to 10 carbon atoms; for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptenyl, cycloheptatrienyl, adamantyl, etc.
The term “halogen” refers to fluorine, chlorine, bromine or iodine.
The term “trihalomethyl” refers to methyl substituted with three same or different halo atoms, such as trifluoromethyl, etc.
In some embodiments in the present application, the pharmaceutically acceptable salt may be a hydrochloride, sulfate, phosphate, maleate, fumarate, citrate, mesylate, p-toluenesulfonate, tartrate, etc.
In some embodiments in the present application, R2 is selected from C1-C3 alkyl, C5-C15 monoalkenyl, C5-C15 dienyl, C5-C15 trienyl, alkynyl, 5-6 membered cycloalkyl, phenyl, 5-6 membered heterocyclyl, 5-6 membered heteroaryl and sterol group; wherein Y and R2 are directly connected, or Y and R2 are connected to form a ring;
R2 is unsubstituted or is substituted by one or more groups selected from C1-C6 alkyl, hydroxyl, halogen, trihalomethyl, carboxyl and phenyl.
Further, in some embodiments of the present application, R2 is selected from methyl, ethyl, propyl, C5 monoalkenyl, C10 dienyl, C15 trienyl, alkynyl, cyclopentyl, cyclohexyl, triazolyl, phenyl, piperidinyl, piperazinyl, pyrrolidinyl, pyridyl, pyrimidyl, sterol group; wherein Y and R2 are directly connected, or Y and R2 are connected to form a ring;
In some embodiments of the present application, R3 is selected from hydrogen, halogen, amino, acetyl, 5-6 membered heterocyclyl, phenyl, biphenyl, naphthyl, 5-6 membered heteroaryl, 5-6 membered cycloalkyl, ester group, carboxyl, amido, trihalomethyl and adamantyl;
Further, in some embodiments of the present application, R3 is selected from hydrogen, halogen, amino, hydroxyl, acetyl, phenyl, biphenyl, naphthyl, cyclopentyl, cyclohexyl, piperidinyl, piperazinyl, pyrrolidinyl, pyridyl, pyrimidyl, ester group, carboxyl, amido, trihalomethyl and adamantyl;
In some embodiments of the present application, the compound has a structure of Formula I or Formula II:
In these embodiments, in the compound of Formula I, X is O or S; Y is —O—, —NH— or
In these embodiments, in the compound of Formula II, X is O or S; Y is —O—, —NH— or
In some embodiments of the present application, when X is O, the compound has a structure of Formula IA or Formula IIA:
In some embodiments of the present application, when X is S, the compound has a structure of Formula IB:
In some embodiments of the present application, when Y is —O—, —NH— or
In some embodiments of the present application, R2′ is selected from
Further, in some embodiments of the present application, in Formula IIA2, R2 is selected from
and R3 is selected from halogen, hydroxyl, phenyl, naphthyl and adamantyl.
As examples, the present application provides a series of compounds selected from the following structures:
In a second aspect of the present application, the present application provides a method for preparing the compounds containing a 2,4-thiazole ring or pharmaceutically acceptable salts or isomers thereof described in the first aspect above, the method comprises:
Technicians in the art can experimentally select suitable reaction conditions according to the preparation method disclosed in the present application, including but not limited to selecting a reaction solvent, selecting reaction temperature, and deciding whether to add a catalyst, etc.
In a third aspect of the present application, the present application provides a pharmaceutical composition or pharmaceutical formulation, comprising the compound or a pharmaceutically acceptable salt or isomer thereof described in the first aspect above.
Alternatively, the pharmaceutical composition or pharmaceutical formulation further comprises a pharmaceutically acceptable excipient or pharmaceutical carrier.
The pharmaceutically acceptable excipient refers to an inert or inactive substance that may be used in the production of a drug or pharmaceutical, which is non-toxic to the subject. The pharmaceutically acceptable excipients include but are not limited to solvents, co-solvents, fillers, lubricants, disintegrants, buffers, stabilizers and preservatives, etc.
The pharmaceutical carrier may be a pharmaceutically acceptable solvent, a suspending agent or a carrier for delivering a compound into an animal or a human body. The carrier may be liquid or solid and is selected according to the planned administration mode. Proteins and liposomes may also be pharmaceutical carriers.
Technicians in the art may use techniques well known in the art to formulate compounds of the present application into pharmaceutical compositions or pharmaceutical formulations. For example, the preparation of pharmaceutical formulations can be carried out according to the guidance of the Modern Pharmaceutical Preparation Series edited by Shenyang Pharmaceutical University. Suitable pharmaceutical excipients, except as mentioned herein, are known in the art, for example, see Handbook of Pharmaceutical Excipients (the fourth edition), authors are Raymond C Rowe and Paul J Sheskey.
In a fourth aspect of the present application, the present application provides a use of the compounds or pharmaceutically acceptable salts or isomers thereof described in the first aspect above, or pharmaceutical compositions or pharmaceutical formulations described in the third aspect above, in the preparation of drugs for the prevention and/or treatment of diseases or conditions related to the anti-activation of the immune system.
Alternatively, the present application provides a use of the compounds or pharmaceutically acceptable salts or isomers thereof described in the above first aspect, or the pharmaceutical compositions or pharmaceutical formulations described in the third aspect above, in the preparation of immunosuppressive drugs.
In embodiments of the present application, the disease or condition is selected from the group consisting of rejection of organ, tissue or cell transplantation, graft-versus-host disease caused by transplantation, autoimmune syndrome, and diseases or conditions associated with cytokine storm.
preferably, the autoimmune syndrome includes lupus, systemic lupus erythematosus, psoriasis, eczema, dermatitis, arthritis, rheumatoid arthritis, spinal arthritis, gouty arthritis or other arthritic conditions, multiple sclerosis, dermatomycosis, antiphospholipid antibody syndrome, struma lymphomatosa, lymphocytic thyroiditis, multiple sclerosis, myasthenia gravis, type 1 diabetes, mellitus, uveitis, episcleritis, scleritis, Kawasaki's disease, uveoretinitis, choroiditis, uveitis associated with Behcet's syndrome, uveoencephalitis, viral encephalomyelitis, chronic allograft vascularopathy, post-infectious autoimmune diseases, rheumatic fever and post-infectious glomerulonephritis, inflammatory and cytoplastic dermatosis, psoriasis, psoriatic arthritis, atopic dermatitis, myopathy, myositis, osteomyelitis, contact dermatitis, dermatitis eczematosa, seborrheic dermatitis, lichen planus, pemphigus, urticaria, angioedema, angiitis, rubeola, acne vulgaris and mast-cell disease;
Further, the disease or condition associated with cytokine storm is cytokine storm syndrome caused by infectious diseases, including but not limited to tumors, inflammation, cytokine storm syndrome caused by infectious diseases such as COVID-19, etc.
In a fifth aspect of the present application, the present application provides a method for preventing and/or treating a disease or condition associated with activating immune system, comprising administering to a subject a therapeutically effective amount of a compound or a pharmaceutically acceptable salt or isomer thereof described in the first aspect above, or a pharmaceutical composition or pharmaceutical formulation described in the third aspect above.
Wherein the subject refers to an animal in need of treatment, observation or experiment, or an animal in treatment, observation or experiment, or an animal that has been subjected to treatment, observation or experiments; the animal particularly refers to a mammal, in particular a human, bovine, rat and mouse.
The therapeutically effective amount refers to an amount of a compound or a pharmaceutically acceptable salt or isomer thereof described in the first aspect above, or an amount of a pharmaceutical composition or pharmaceutical formulation comprising the compound or a pharmaceutically acceptable salt or isomer thereof, which may cause a biological or medical response of a tissue system, an animal or a person pursued by researchers, veterinarians, doctors, or other medical personnel, including alleviating or partially alleviating symptoms of the treated disease, syndrome, condition or disorder.
Compared with the existing technology, the present application has the following advantages:
The present application provides a series of novel compounds containing a 2,4-thiazole ring, which are synthesized by a simple and efficient method with high yields and low toxicity. These compounds have significant inhibitory effects on T lymphocytes and B lymphocytes and can be used for the preparation of immunosuppressive drugs.
The accompanying drawings to the specification, which form part of the present application, are used to provide a further understanding of the present application, and the illustrative embodiments of the present application and the description thereof are used to explain the present application and are not unduly limiting the present application. Hereinafter, embodiments of the present application are described in detail with reference to the accompanying drawings, wherein:
The present application is further described below with reference to specific embodiments. It should be understood that these embodiments are intended to illustrate the present application only and not to limit the scope of the present application. Experimental methods for which specific conditions are not indicated in the following embodiments generally follow conventional conditions or follow the conditions recommended by the manufacturer.
Unless otherwise defined, all professional and scientific terms used in the text have the same meaning as those familiar to those skilled in the art. Reagents or raw materials used in the present application are available through conventional means. Unless otherwise specified, reagents or raw materials used in the present application are used in a conventional manner in the field or in accordance with product specifications. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present disclosure. The preferred embodiments described herein are exemplary only.
In the examples, 1H NMR and 13C NMR were recorded by Avance III-400 or Avance III-600 NMR instrument with chemical shifts expressed as δ (ppm); mass spectrometry was recorded by MS-LCQ-DECA ion trap mass spectrometer (ESI/LR) and MS-Q-TOF quadrupole-time-of-flight mass spectrometry (ESI-HR); 200-300 mesh silica gel (Sinopharm Chemical Reagent Co., Ltd.) was used for compound separation.
The present invention provides a series of compounds containing a 2,4-thiazole ring, and the compounds can be prepared by the following method:
Preparation route I:
The reaction step comprises:
Wherein preparing a compound of Formula IB by oxidation sulfur exchange of a compound of formula IA, which reacts as follows:
Preparation route II:
Specifically, the preparation method and activity of exemplary compounds are illustrated in the following examples. Those skilled in the art can prepare more compounds that conform to the generic structure of the present application based on the disclosure of the present application.
Amines or alcohols containing different substituents were used as compound 10, and different compounds IA were prepared according to steps similar to those described previously in this example, and the structures of compound 10 and the prepared compounds IA are listed in Table 1:
Compound IA-1: yellow solid, the yield was 87.2%. 1H NMR (400 MHz, CDCl3) δ9.29-9.21 (m, 2H), 8.83 (dd, J=15.0, 3.1 Hz, 1H), 8.64-8.60 (m, 1H), 8.47-8.39 (m, 1H), 7.88 (s, 1H), 7.65 (dt, J=15.0, 3.1 Hz, 1H), 7.28 (td, J=14.9, 4.7 Hz, 2H), 4.74-4.66 (m, 1H), 4.04 (dt, J=19.8, 13.3 Hz, 1H), 3.81 (dt, J=24.8, 13.2 Hz, 1H), 2.24 (dtd, J=12.0, 8.8, 1.6 Hz, 1H), 2.13-1.88 (m, 3H).
Compound IA-2: yellow solid, the yield was 85.3%. 1H NMR (400 MHz, CDCl3) δ9.25 (dd, J=15.0, 2.9 Hz, 1H), 8.98 (s, 1H), 8.83 (dd, J=15.0, 2.9 Hz, 1H), 8.62(m, 1H), 8.43 (m, 1H), 8.38 (s, 1H), 7.65 (dt, J=15.0, 3.1 Hz, 1H), 7.29 (m, 3H), 4.23 (s, 2H).
Compound IA-3: yellow solid, the yield was 86.5%. 1H NMR (400 MHz, CDCl3) δ8.76 (d, J=7.0 Hz, 1H), 8.71 (dd, J=5.5, 3.5 Hz, 2H), 8.67 (s, 1H), 8.56 (dd, 1H), 8.16 (s, 1H), 7.93 (s, 1H), 7.81 (d, J=7.8 Hz, 1H), 7.33(t, 1H), 7.01 (t, 1H), 4.72 (d, J=5.9 Hz, 2H), 2.46 (s, J=1.2 Hz, 1H).
Compound IA-4: yellow solid, the yield was 89.8%. 1H NMR (400 MHz, DMSO-d6) δ9.38 (dd, J=7.1, 1.7 Hz, 1H), 8.92 (dd, J=4.1, 1.7 Hz, 1H), 8.88 (s, 1H), 8.55 (s, 1H), 7.35 (dd, J=7.0, 4.2 Hz, 1H), 5.51 (td, 1H), 4.88 (d, J=7.3 Hz, 2H), 1.83 (dd, J=3.0, 1.3 Hz, 6H).
Compound IA-5: yellow solid, the yield was 82.3%. 1H NMR (400 MHz, Chloroform-d) δ8.75 (d, J=7.0 Hz, 2H), 8.72-8.68 (m, 1H), 8.14 (s, 1H), 7.87 (t, J=6.4 Hz, 1H), 7.45 (m, J=8.0 Hz, 1H), 6.99 (dd, J=7.1, 4.2 Hz, 1H), 6.84 (m, J=9.6, 9.0 Hz, 2H), 4.69 (d, J=6.3 Hz, 2H).
Compound IA-6: yellow solid, the yield was 85.5%. 1H NMR (400 MHz, CDCl3) δ8.89 (s, 1H), 8.74 (dd, J=7.0, 1.8 Hz, 1H), 8.67 (dd, J=4.1, 1.7 Hz, 1H), 8.17 (s, 1H), 6.96 (dd, J=7.0, 4.1 Hz, 1H), 5.49 (t, J=6.7 Hz, 1H), 5.08 (t, J=6.1 Hz, 1H), 4.90 (d, J=7.1 Hz, 2H), 2.08 (td, J=9.8, 7.5, 4.3 Hz, 5H), 1.76 (s, 3H), 1.66 (s, 3H), 1.58 (s, 3H).
Compound IA-7: yellow solid, the yield was 85.4%. 1H NMR (400 MHz, CDCl3) δ9.25 (m,2H), 8.83 (dd, J=15.0, 3.1 Hz, 1H), 7.88 (s, 1H), 7.28 (t, J=15.0 Hz, 1H), 5.34(dddt, J=14.3, 10.3, 4.0, 2.0 Hz, 1H), 5.15 (dddt, J=14.3, 10.3, 4.0, 2.0 Hz, 1H), 4.67 (d, J=12.4 Hz, 2H), 2.02 (m, 4H), 1.82 (d, J=2.0 Hz, 3H), 1.70 (d, J=2.0 Hz, 3H), 1.66 (d, J=2.0 Hz, 3H).
Compound IA-8: yellow solid, the yield was 79.4%. 1H NMR (400 MHz, CDCl3) δ9.24 (m, 2H), 8.83 (dd, J=7.5, 1.4 Hz, 1H), 7.89 (d, J=12.1 Hz, 2H), 7.75 (d, J=1.4 Hz, 1H), 7.25 (m, 3H), 4.23 (s, 2H).
Compound IA-9: yellow solid, the yield was 87.3%. 1H NMR (400 MHz, CDCl3) δ8.82 (s, 1H), 8.77 (dd, J=7.1, 1.8 Hz, 1H), 8.71 (dd, J=6.8, 1.9 Hz, 3H), 8.64 (dd, J=4.1, 1.7 Hz, 2H), 8.29 (s, 2H), 8.09 (s, 1H), 7.84 (s, 2H), 7.37-7.25 (m, 10H), 7.23-7.15 (m, 7H), 6.99 (dd, J=7.0, 4.1 Hz, 1H), 6.94 (dd, J=7.0, 4.1 Hz, 2H), 6.01 (dd, J=7.7, 2.2 Hz, 2H), 5.44 (dd, J=7.8, 4.4 Hz, 1H), 4.44 (dt, J=12.2, 6.6 Hz, 1H), 4.29 (dt, J=11.6, 6.9 Hz, 1H), 3.98 (dd, J=8.6, 5.6 Hz, 4H), 2.42 (ddt, J=14.6, 7.4, 3.4 Hz, 4H), 2.16-1.86 (m, 11H).
Compound IA-10: yellow solid, the yield was 89.5%. 1H NMR (400 MHz, DMSO-d6) δ9.37 (dd, J=7.0, 1.6 Hz, 1H), 9.11 (d, J=2.3 Hz, 1H), 9.02 (s, 1H), 8.91 (dd, J=4.1, 1.6 Hz, 1H), 8.49 (s, 1H), 8.45-8.40 (m, 1H), 8.37 (dd, J=8.3, 2.1 Hz, 1H), 7.52 (dd, J=8.3, 4.7 Hz, 1H), 7.34 (dd, J=7.0, 4.1 Hz, 1H).
Compound IA-11: yellow solid, the yield was 82.6%. 1H NMR (400 MHz, CDCl3) δ8.66 (d, J=5.2 Hz, 2H), 8.64-8.59 (m, 1H), 8.08 (s, 1H), 7.82 (s, 1H), 7.33 (d, J=7.5 Hz, 2H), 7.28 (t, J=7.2 Hz, 2H), 7.24-7.18 (m, 1H), 6.94-6.87 (m, 1H), 4.62 (d, J=5.9 Hz, 2H).
Compound IA-12: yellow solid, the yield was 84.9%. 1H NMR (400 MHz, Chloroform-d) δ8.72 (d, J=7.0 Hz, 1H), 8.65 (s, 2H), 8.52 (s, 1H), 8.47 (d, J=4.1 Hz, 1H), 8.06 (s, 1H), 7.61 (d, J=7.5 Hz, 2H), 7.24 (d, J=6.7 Hz, 1H), 6.98-6.92 (m, 1H), 3.71 (q, J=6.8 Hz, 2H), 2.97 (t, J=7.1 Hz, 2H).
Compound IA-13: yellow solid, the yield was 87.9%. 1H NMR (400 MHz, Chloroform-d) δ8.74-8.68 (m, 2H), 8.63 (dt, J=6.0, 1.7 Hz, 2H), 8.55 (dd, J=4.0, 1.8 Hz, 1H), 8.51-8.46 (m, 2H), 8.45-8.39 (m, 2H), 8.09 (s, 1H), 8.02 (s, 1H), 7.94 (s, 1H), 7.21 (d, J=5.4 Hz, 2H), 7.13-7.07 (m, 2H), 6.91 (dd, J=7.0, 4.1 Hz, 1H), 6.86 (dd, J=7.0, 4.0 Hz, 1H), 6.07 (dd, J=7.9, 2.1 Hz, 1H), 5.29 (dd, J=8.1, 5.0 Hz, 1H), 4.38 (dt, J=11.7, 6.7 Hz, 1H), 4.24 (dt, J=11.6, 7.0 Hz, 1H), 3.96-3.85 (m, 2H), 2.39 (tdt, J=15.0, 12.6, 7.2 Hz, 2H), 1.96 (dtt, J=12.6, 9.9, 7.6, 3.6 Hz, 5H), 1.80 (dtd, J=22.6, 11.7, 11.3, 8.0 Hz, 2H).
Compound IA-14: yellow solid, the yield was 81.2%. 1H NMR (400 MHz, CDCl3) δ8.76 (dd, J=7.1, 1.7 Hz, 1H), 8.66 (d, J=4.0 Hz, 2H), 8.12 (s, 1H), 7.00 (dd, J=7.0, 4.1 Hz, 1H), 4.07 (q, J=9.1 Hz, 2H).
Compound IA-15: yellow solid, the yield was 86.3%. 1H NMR (400 MHz, DMSO-d6) δ9.29-9.25 (m, 2H), 8.88-8.80 (m, 3H), 8.66 (s, 1H), 8.26 (s, 1H), 8.23 (s, 1H), 7.26-7.21(m, 2H), 5.41 (t, J=7.9 Hz, 1H), 5.18 (d, J=3.8 Hz, 1H), 5.13 (d, J=3.4 Hz, 1H), 4.58 (t, J=8.4 Hz, 1H), 4.41 (s, 1H), 4.32 (q, J=3.8 Hz, 1H), 4.17-3.99 (m, 3H), 3.74 (dt, J=12.3, 2.1 Hz, 1H), 3.69-3.59 (m, 5H), 3.51 (s, 3H), 2.37 (td, J=10.7, 8.7, 3.2 Hz, 1H), 2.26-2.07 (m, 2H), 1.96 (ddd, J=13.2, 9.2, 4.4 Hz, 1H).
Compound IA-16: yellow solid, the yield was 86.4%. 1H NMR (400 MHz, CDCl3) δ9.29-9.21 (m, 2H), 8.83 (dd, J=15.0, 3.1 Hz, 1H), 7.88 (s, 1H), 7.28 (t, J=15.0 Hz, 1H), 4.87 (t, J=6.2 Hz, 1H), 3.68-3.42 (m, 5H), 2.35-2.17 (m, 1H), 2.09-1.90 (m, 1H), 1.87-1.52 (m, 4H).
Compound IA-17: yellow solid, the yield was 83.5%. 1H NMR (400 MHz, CDCl3) δ9.28-9.21 (m, 2H), 8.83 (dd, J=7.5, 1.4 Hz, 1H), 7.88 (s, 1H), 7.28 (t, J=7.5 Hz, 1H), 7.24-7.06 (m, 3H), 4.59 (t, J=4.6 Hz, 1H), 4.06 (dt, J=13.0, 6.6 Hz, 1H), 3.88-3.79 (m, 1H), 2.30-2.15 (m, 3H), 2.07-1.97 (m, 1H).
Compound IA-18: yellow solid, the yield was 82.3%. 1H NMR (400 MHz, CDCl3) δ8.87 (s, 1H), 8.78 (d, J=3.2 Hz, 1H), 8.45 (dd, J=6.2, 2.6 Hz, 1H), 8.33 (s, 1H), 7.28 (t, J=15.0 Hz, 1H), 3.74 (dd, J=12.6, 6.3 Hz, 2H), 2.52-2.36 (m, 2H).
Compound IA-19: yellow solid, the yield was 89.3%. 1H NMR (400 MHz, MeOD) δ9.06 (dd, J=7.0, 1.4 Hz, 1H), 8.81 (s, 1H), 8.78 (dd, J=4.0, 1.4 Hz, 1H), 8.19 (s, 1H), 7.19 (dd, J=7.0, 4.1 Hz, 1H), 4.35-4.23 (m, 1H), 3.86 (dd, J=13.9, 3.9 Hz, 1H), 3.56 (dd, J=13.9, 8.5 Hz, 1H).
Compound IA-20: yellow solid, the yield was 85.7%. 1H NMR (400 MHz, CDCl3) δ8.81-8.73 (m, 2H), 8.70 (dd, J=4.0, 1.5 Hz, 1H), 8.10 (s, 1H), 7.40 (d, J=8.1 Hz, 1H), 6.99 (dd, J=7.0, 4.1 Hz, 1H), 3.96 (tdt, J=11.9, 8.2, 3.9 Hz, 1H), 2.23 (d, J=10.3 Hz, 2H), 2.03 (m, J=10.0 Hz, 4H), 1.51 (dt, J=14.4, 12.3 Hz, 2H), 1.42-1.28 (m, 2H).
Compound IA-21: yellow solid, the yield was 83.4%. 1H NMR (400 MHz, CDCl3) δ8.77 (dd, J=7.1, 1.8 Hz, 1H), 8.70-8.64 (m, 2H), 8.52 (d, J=4.9 Hz, 1H), 8.06 (s, 1H), 7.65 (td, J=7.7, 1.8 Hz, 1H), 7.29 (d, J=5.7 Hz, 1H), 7.21-7.15 (m, 1H), 6.99 (dd, J=7.1, 4.1 Hz, 1H), 3.82 (t, J=7.0 Hz, 2H), 3.34 (s, 1H), 3.12 (t, J=7.0 Hz, 2H).
Compound IA-22: yellow solid, the yield was 88.6%. 1H NMR (400 MHz, DMSO-d6) δ9.31 (dd, J=7.0, 1.6 Hz, 1H), 9.15 (t, J=6.1 Hz, 1H), 9.10 (s, 1H), 8.86 (dd, J=4.1, 1.6 Hz, 1H), 8.81 (s, 2H), 8.27 (s, 1H), 7.29 (dd, J=7.0, 4.1 Hz, 1H), 4.55 (d, J=6.1 Hz, 2H).
Compound IA-23: yellow solid, the yield was 85.5%. 1H NMR (400 MHz, CDCl3) δ8.77 (dt, J=6.2, 3.1 Hz, 1H), 8.74-8.68 (m, 2H), 8.34 (d, J=1.9 Hz, 1H), 8.12 (t, J=4.7 Hz, 1H), 8.00 (s, 1H), 7.44 (dt, J=8.6, 6.8 Hz, 2H), 7.00 (dt, J=8.9, 4.5 Hz, 1H), 3.41-3.32 (m, 4H), 3.30-3.22 (m, 4H).
Compound IA-24: yellow solid, the yield was 73.5%. 1H NMR (400 MHz, CDCl3) δ8.79 (s, 1H), 8.76 (dd, J=7.0, 1.5 Hz, 1H), 8.71 (dd, J=3.9, 1.4 Hz, 1H), 8.11 (s, 1H), 7.47 (s, 1H), 7.00 (dd, J=7.0, 4.1 Hz, 1H), 5.34 (t, J=6.6 Hz, 1H), 5.10 (t, J=6.6 Hz, 1H), 4.10 (t, J=6.2 Hz, 2H), 2.19-2.09 (m, 2H), 2.08-2.02 (m, 2H), 1.75 (s, 3H), 1.68 (s, 3H), 1.61 (s, 3H).
Compound IA-25: yellow solid, the yield was 82.2%. 1H NMR (400 MHz, CDCl3) δ8.63 (s, 1H), 8.61-8.49 (m, 3H), 8.10 (s, 1H), 7.66 (t, J=8.2 Hz, 2H), 7.31 (dd, J=7.8, 4.7 Hz, 1H), 3.77 (q, J=7.0 Hz, 2H), 3.02 (t, J=7.2 Hz, 2H), 2.46 (s, 3H).
Compound IA-26: yellow solid, the yield was 83.5%. 1H NMR (400 MHz, CDCl3) δ8.78-8.71 (m, 2H), 8.69 (dd, J=4.0, 1.7 Hz, 1H), 8.59 (d, J=4.2 Hz, 1H), 8.13 (s, 1H), 8.05 (d, J=7.9 Hz, 1H), 7.99 (t, J=6.3 Hz, 1H), 7.47 (dd, J=7.9, 4.7 Hz, 1H), 6.99 (dd, J=7.0, 4.1 Hz, 1H), 4.88 (d, J=6.4 Hz, 2H).
Compound IA-27: yellow solid, the yield was 79.0%. 1H NMR (400 MHz, CDCl3) δ8.65 (dt, J=11.8, 6.5 Hz, 4H), 8.09 (s, 1H), 8.03 (t, J=6.4 Hz, N-H), 7.85 (d, J=8.2 Hz, 1H), 7.56 (d, J=8.0 Hz, 1H), 6.93 (dd, J=7.0, 4.1 Hz, 1H), 4.68 (d, J=4.4 Hz, 2H).
Compound IA-28: yellow solid, the yield was 81.3%. 1H NMR (400 MHz, DMSO-d6) δ9.29 (dd, J=7.1, 1.7 Hz, 1H), 9.13 (t, J=6.3 Hz, 1H), 8.84 (dd, J=4.1, 1.7 Hz, 1H), 8.81 (s, 1H), 8.59 (dd, J=8.9, 2.0 Hz, 2H), 8.26 (s, 1H), 8.02 (t, J=2.1 Hz, 1H), 7.27 (dd, J=7.0, 4.1 Hz, 1H), 4.54 (d, J=6.2 Hz, 2H).
Compound IA-29: yellow solid, the yield was 73.3%. 1H NMR (400 MHz, CDCl3) δ8.79-8.68 (m, 3H), 8.22 (d, J=1.2 Hz, 1H), 8.16 (s, 1H), 8.03 (t, J=5.9 Hz, 1H), 7.56 (dd, J=8.5, 1.7 Hz, 1H), 7.01 (dd, J=7.0, 4.1 Hz, 1H), 4.67 (d, J=6.4 Hz, 2H).
Compound IA-30: yellow solid, the yield was 83.5%. 1H NMR (400 MHz, CDCl3) δ8.79-8.68 (m, 1H), 8.23 (s, 1H), 8.17 (d, J=5.8 Hz, 1H), 7.94 (s, 1H), 7.87 (d, J=5.4 Hz, 1H), 7.00 (dd, J=7.0, 4.1 Hz, 1H), 6.93 (d, J=4.1 Hz, 1H), 4.68 (d, J=6.3 Hz, 1H).
Compound IA-31: yellow solid, the yield was 83.6%. 1H NMR (400 MHz, CDCl3) δ8.76 (d, J=7.0 Hz, 1H), 8.72 (d, J=4.4 Hz, 2H), 8.42 (s, 1H), 8.16 (s, 1H), 7.93 (s, 1H), 7.74 (dd, J=8.1, 1.8 Hz, 1H), 7.30 (t, J=7.3 Hz, 1H), 7.01 (dd, J=6.9, 4.1 Hz, 1H), 4.68 (d, J=6.3 Hz, 2H).
Compound IA containing different substituents were used as starting materials, and different compounds IB were prepared according to steps similar to those described previously in this example, and the structures of compound IA and the prepared compounds IB are listed in the below Table:
Compound IB-1: yellow solid, the yield was 69.2%. 1H NMR (400 MHz, CDCl3) δ9.25 (dd, J=15.0, 3.1 Hz, 1H), 8.83 (dd, J=15.0, 3.1 Hz, 1H), 8.62 (m, 1H), 8.43 (m, 2H), 7.88 (s, 1H), 7.65(dt, J=15.02,3.06 1H), 7.28 (td, J=14.9, 4.7 Hz, 2H), 4.40 (s, 2H).
Compound IB-2: yellow solid, the yield was 77.3%. 1H NMR (400 MHz, CDCl3) δ9.25 (dd, J=7.4, 1.5 Hz, 1H), 8.83 (dd, J=7.5, 1.4 Hz, 1H), 8.50 (d, J=1.3 Hz, 1H), 8.44 (s, 1H), 8.13 (dd, J=7.5, 1.3 Hz, 1H), 7.88 (s, 1H), 7.27 (dt, J=9.7, 7.5 Hz, 2H), 7.09 (dt, J=7.5, 1.5 Hz, 1H).
Compound IB-3: yellow solid, the yield was 73.7%. 1H NMR (400 MHz, CDCl3) δ9.41 (s, 1H), 9.25 (dd, J=15.0, 3.1 Hz, 1H), 8.83 (dd, J=15.0, 3.1 Hz, 1H), 8.61 (s, 2H), 8.54 (s, 1H), 8.44 (s, 1H), 7.88 (s, 1H), 7.26 (d, J=15.0 Hz, 1H), 4.40 (s, 2H).
Compound IB-4: yellow solid, the yield was 75.0%. 1H NMR (400 MHz, CDCl3) δ9.25 (dd, J=7.4, 1.5 Hz, 1H), 9.00 (s, 1H), 8.83 (dd, J=7.5, 1.4 Hz, 1H), 8.42(m, 3H), 7.88 (s, 1H), 7.46 (dt, J=7.5, 1.4 Hz, 1H), 7.28 (t, J=7.5 Hz, 1H), 7.16 (t, J=7.5 Hz, 1H), 3.54 (t, J=7.6 Hz, 2H), 2.81 (t, J=7.6 Hz, 2H).
Amines or alcohols containing different substituents were used as compound 19, and different compounds IIA were prepared according to steps similar to those described previously in this example, and the structures of compound 19 and the prepared compounds IIA are listed in Table 3:
Compound IIA-1: yellow solid, the yield was 65.2%. 1H NMR (400 MHz, CDCl3) δ10.13 (s, 1H), 9.41 (s, 1H), 8.90 (d, J=3.0 Hz, 1H), 8.62 (s, 2H), 8.47 (dd, J=28.3, 5.9 Hz, 3H), 8.30 (d, J=4.9 Hz, 2H), 8.18 (s, 1H), 7.73 (d, J=7.9 Hz, 1H), 7.54 (d, J=7.8 Hz, 1H), 7.48-6.88 (m, 10H), 5.83 (d, J=7.2 Hz, 1H), 5.46-5.27 (m, 1H), 4.46-4.07 (m, 1H), 4.07-3.78 (m, 3H), 3.00-2.19 (m, 2H), 1.93 (tddd, J=31.4, 25.2, 12.5, 6.2 Hz, 5H), 1.69 (dt, J=16.2, 11.2 Hz, 1H).
Compound IIA-2: yellow solid, the yield was 79.5%. 1H NMR (400 MHz, CDCl3) δ9.73 (s, 1H), 8.26 (dd, J=14.7, 3.3 Hz, 1H), 8.22 (s, 1H), 7.97 (s, 1H), 7.53 (dd, J=14.6, 3.3 Hz, 1H), 7.27 (td, J=14.9, 3.4 Hz, 1H), 7.18 (td, J=14.8, 3.3 Hz, 1H), 5.33 (tdd, J=12.5, 4.0, 2.0 Hz, 1H), 5.14 (tdd, J=12.5, 1.9, 1.1 Hz, 2H), 4.66 (d, J=12.3 Hz, 2H), 2.18 (m, 6H), 1.96 (m, 2H), 1.82 (d, J=1.8 Hz, 3H), 1.70 (d, J=2.0 Hz, 3H), 1.66 (d, J=2.0 Hz, 6H).
Compound IIA-3: yellow solid, the yield was 83.6%. 1H NMR (400 MHz, DMSO-d6) δ12.34 (d, J=3.3 Hz, 1H), 9.07 (d, J=3.3 Hz, 1H), 8.83 (s, 1H), 8.37-8.28 (m, 1H), 7.67-7.51 (m, 1H), 7.28 (q, J=4.8 Hz, 2H), 5.02 (d, J=2.5 Hz, 2H), 3.64 (s, 1H).
Compound IIA-4: yellow solid, the yield was 84.8%. 1H NMR(400 MHz, CDCl3): δ9.75 (s, 1H), 8.31-8.23 (m, 2H), 7.98 (s, 1H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), δ7.28 (td, J=14.9, 3.4 Hz, 1H), 7.19 (td, J=14.8, 3.2 Hz, 1H), 5.34 (dddt, J=14.4, 12.5, 4.0, 2.0 Hz, 1H), 4.67 (d, J=12.4 Hz, 2H), 1.82 (d, J=2.0 Hz, 3H), 1.70 (d, J=2.0 Hz, 3H).
Compound IIA-5: yellow solid, the yield was 90.0%. 1H NMR (400 MHz, CDCl3) δ9.75 (s, 1H), 8.26 (m, 2H), 7.98 (s, 1H), 7.54 (dd, J=14.6, 3.4 Hz, 1H), 7.23 (dtd, J=47.6, 14.8, 3.3 Hz, 2H), 5.34 (tdd, J=12.5, 4.0, 2.0 Hz, 1H), 5.15 (dddt, J=14.3, 10.3, 4.0, 2.0 Hz, 1H), 4.67 (d, J=12.4 Hz, 2H), 2.12 (m, 4H), 1.82 (d, J=2.0 Hz, 3H), 1.70 (d, J=2.0 Hz, 3H), 1.66 (d, J=2.0 Hz, 3H).
Compound IIA-6: yellow solid, the yield was 71.2%. 1H NMR (400 MHz, CDCl3) δ9.47-9.35 (m, 2H), 8.66 (s, 1H), 8.62 (s, 1H), 8.48 (d, J=3.6 Hz, 1H), 8.33 (d, J=6.0 Hz, 1H), 7.79 (d, J=7.7 Hz, 1H), 7.57 (d, J=6.4 Hz, 1H), 7.38 (dd, J=7.4, 4.9 Hz, 1H), 7.29 (dd, J=9.1, 5.3 Hz, 2H), 4.60 (d, J=6.0 Hz, 2H).
Compound IIA-7: yellow solid, the yield was 81.3%. 1H NMR (400 MHz, CDCl3) δ9.75 (s, 1H), 8.27 (m, 2H), 7.98 (s, 1H), 7.54 (dd, J=7.5, 1.6 Hz, 1H), 7.28 (td, J=7.5, 1.5 Hz, 1H), 7.19 (td, J=7.5, 1.5 Hz, 1H), 5.27 (t, J=6.2 Hz, 1H), 4.84 (p, J=7.2 Hz, 1H), 4.36 (dt, J=9.9, 7.3 Hz, 1H), 3.50 (dd, J=12.5, 2.0 Hz, 1H), 3.25 (dd, J=12.5, 2.0 Hz, 1H), 2.78 (ddd, J=12.4, 7.1, Hz, 1H), 2.19 (ddd, J=12.3, 8.4, 6.2 Hz, 1H), 2.08 (dq, J=12.7, 6.9 Hz, 1H), 1.59 (m, 18H), (m, 12H), 0.80 (d, J=6.4 Hz, 3H).
Compound IIA-8: yellow solid, the yield was 84.5%. 1H NMR (400 MHz, CDCl3) δ9.75 (s, 1H), 8.27 (dd, J=15.0, 3.1 Hz, 2H), 7.98 (s, 1H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), 7.23 (dtd, J=47.4, 14.9, 3.2 Hz, 2H), 5.27 (tt, J=12.5, 2.0 Hz, 1H), 4.84 (p, J=14.7 Hz, 1H), 2.47 (ddd, J=24.7, 14.7, 1.9 Hz, 1H), 2.21 (m, 3H), 1.53 (m, 23H), 90 (m, 15H), 0.49 (dt, J=22.7, 18.5 Hz, 1H).
Compound IIA-9: yellow solid, the yield was 83.9%. 1H NMR (400 MHz, CDCl3) δ9.75 (s, 1H), 8.27 (q, J=3.2 Hz, 2H), 7.98 (s, 1H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), 7.23 (dtd, J=47.4, 14.8, 3.2 Hz, 2H), 5.27 (tt, J=12.3, 2.0 Hz, 1H), 4.82 (p, J=14.7 Hz, 1H), 2.45 (ddd, J=24.9, 14.7, 2.0 Hz, 1H), 2.23 (m, 3H), 1.42 (m, 40H), 0.49 (dt, J=22.5, 18.4 Hz, 1H).
Compound IIA-10: yellow solid, the yield was 89.4%. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.27 (dd, J=6.9, 1.9 Hz, 2H), 7.98 (s, 1H), 7.54 (dd, J=7.5, 1.6 Hz, 1H), 7.28 (td, J=7.5, 1.5 Hz, 1H), 7.19 (td, J=7.5, 1.5 Hz, 1H), 6.67 (s, 1H), 5.27 (t, J=6.2 Hz, 1H), 4.08 (p, J=7.6 Hz, 1H), 3.79 (dt, J=10.4, 8.0 Hz, 1H), 3.50 (dd, J=12.4, 8.6 Hz, 1H), 3.25 (dd, J=12.4, 8.6 Hz, 1H), 2.47 (ddd, J=12.5, 7.9, 0.8 Hz, 1H), 2.18 (m, 2H), 1.64 (m, 19H), 0.89 (m, 10H), 0.77 (m, 4H).
Compound IIA-11: yellow solid, the yield was 84.7%. 1H NMR (400 MHz, DMSO-d6) δ12.41 (d, J=3.3 Hz, 1H), 9.11 (d, J=3.3 Hz, 1H), 8.94 (s, 1H), 8.33 (dt, J=7.8, 3.0 Hz, 1H), 7.60(dd, 1H), 7.52 (dd, 1H), 7.45 (t, J=7.4 Hz, 2H), 7.41-7.37 (m, 1H), 7.36-7.32 (m, 5H), 7.23 (dd, J=7.7, 1.4 Hz, 1H), 5.51 (s, 2H), 2.28 (s, 3H).
Compound IIA-12: yellow solid, the yield was 78.9%. 1H NMR (400 MHz, MeOD) δ11.72 (s, 1H), 9.28 (d, J=2.6 Hz, 1H), 8.36 (s, 1H), 8.26 (dd, J=5.6, 3.4 Hz, 1H), 7.45-7.38 (m, 1H), 7.25-7.13 (m, 2H), 3.46 (t, J=5.7 Hz, 2H), 3.38 (t, J=5.8 Hz, 2H), 1.89 (s, 4H).
Compound IIA-13: yellow solid, the yield was 83.5%. 1H NMR (400 MHz, MeOD) δ9.20 (d, J=1.8 Hz, 1H), 8.54 (d, J=1.4 Hz, 1H), 8.41-8.36 (dd, 1H), 7.57-7.48 (dd, 1H), 7.30 (m, J=6.3 Hz, 2H), 4.82 (t, J=3.8 Hz, 1H), 4.13 (dd, J=11.4, 4.1 Hz, 1H), 4.04 (dd, J=11.4, 3.6 Hz, 1H), 3.85 (s, 3H).
Compound IIA-14: yellow solid, the yield was 88.6%. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.27 (dd, J=14.7, 3.4 Hz, 1H), 8.18 (s, 1H), 7.98 (s, 1H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), 7.40 (s, 1H), 7.29(m, 2H), 7.19 (td, J=14.8, 3.2 Hz, 1H), 6.96 (m, 1H), 6.87 (td, J=16.0, 3.0 Hz, 1H), 4.23 (s, 2H).
Compound IIA-15: yellow solid, the yield was 87.1%. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.27 (m, 2H), 7.98 (s, 1H), 7.54 (dd, J=7.5, 1.6 Hz, 1H), 7.28 (td, J=7.5, 1.5 Hz, 1H), 7.19 (td, J=7.5, 1.5 Hz, 1H), 6.66 (s, 1H), 3.18 (t, J=7.7 Hz, 2H), 2.51(m, 4H), 2.36 (t, J=7.6 Hz, 2H), 1.73 (m, 2H), 1.68(m, 4H).
Compound IIA-16: yellow solid, the yield was 82.3%. 1H NMR (400 MHz, DMSO-d6) δ12.35 (s, N-H), 9.45 (s, 1H), 9.36 (t, J=6.2 Hz, 1H), 8.68 (s, 1H), 8.39-8.31 (m, 1H), 7.64 (d, J=1.8 Hz, 1H), 7.61-7.55 (m, 1H), 7.49-7.38 (m, 2H), 7.36-7.25 (m, 2H), 4.62 (d, J=6.0 Hz, 2H).
Compound IIA-17: yellow solid, the yield was 88.4%. 1H NMR (400 MHz, DMSO) δ12.45 (s, 1H), 10.62 (s, 1H), 9.53 (d, J=3.2 Hz, 1H), 9.02 (d, J=2.3 Hz, 1H), 8.85 (s, 1H), 8.39 (dd, J=4.7, 1.2 Hz, 1H), 8.37-8.31 (m, 1H), 8.31-8.24 (m, 1H), 7.64-7.55 (m, 1H), 7.47 (dd, J=8.3, 4.7 Hz, 1H), 7.35-7.26 (m, 2H).
Compound IIA-18: yellow solid, the yield was 88.8%. 1H NMR (400 MHz, CDCl3) δ10.39 (s, 1H), 9.44 (s, 2H), 8.92 (d, J=3.1 Hz, 1H), 8.50 (d, J=7.8 Hz, 1H), 8.42 (d, J=7.6 Hz, 2H), 8.25 (s, 1H), 8.05 (s, 2H), 7.82 (d, J=3.2 Hz, 2H), 7.66-6.88 (m, 25H), 5.66 (d, J=6.9 Hz, 2H), 5.40 (dd, J=7.5, 4.7 Hz, 1H), 4.19 (ddd, J=18.0, 12.3, 6.5 Hz, 2H), 4.08 (dd, J=29.5, 8.0 Hz, 1H), 4.03-3.81 (m, 3H), 2.70-2.07 (m, 4H), 2.07-1.49 (m, 9H).
Compound IIA-19: yellow solid, the yield was 87.9%. 1H NMR (400 MHz, MeOD) δ9.34 (s, 1H), 8.53 (s, 1H), 8.39 (d, J=7.3 Hz, 1H), 7.52 (d, J=7.0 Hz, 1H), 7.42 (d, J=7.5 Hz, 2H), 7.36 (t, J=7.3 Hz, 2H), 7.29 (t, 3H), 4.69 (s, 2H).
Compound IIA-20: yellow solid, the yield was 90.1%. 1H NMR (400 MHz, DMSO) δ12.41 (s, 1H), 9.39 (s, 1H), 8.89 (t, J=5.3 Hz, 1H), 8.60 (s, 1H), 8.55 (s, 1H), 8.47 (d, J=4.5 Hz, 1H), 8.35 (d, J=7.2 Hz, 1H), 7.79 (d, J=7.5 Hz, 1H), 7.60 (d, J=7.2 Hz, 1H), 7.40 (t, 1H), 7.31(m, 2H), 3.63 (dd, J=13.0, 6.5 Hz, 2H), 2.97 (dd, 2H).
Compound IIA-21: yellow solid, the yield was 82.3%. 1H NMR (400 MHz, CDCl3) δ10.60 (s, 1H), 9.58 (s, 2H), 8.97 (d, J=3.1 Hz, 1H), 8.64 (d, J=5.0 Hz, 4H), 8.54 (t, J=5.9 Hz, 3H), 8.41 (d, J=7.3 Hz, 2H), 8.32 (s, 3H), 7.61-7.21 (m, 16H), 7.17 (d, J 32 3.1 Hz, 2H), 5.57 (d, J=7.5 Hz, 2H), 5.48-5.28 (m, 4H), 4.70-3.96 (m, 5H), 3.96-3.82 (m, 2H), 2.38 (dtd, J=19.9, 12.7, 7.4 Hz, 3H), 2.16-1.76 (m, 9H), 1.75 (d, J=6.7 Hz, 1H).
Compound IIA-22: yellow solid, the yield was 84.0%. 1H NMR (400 MHz, CDCl3) δ9.68 (s, 1H), 8.22 (dd, J=14.7, 3.3 Hz, 1H), 8.15 (s, 1H), 7.93 (s, 1H), 7.49 (dd, J=14.6, 3.4 Hz, 1H), 7.19 (dtd, J=47.3, 14.8, 3.3 Hz, 2H), 4.37 (t, J=16.3 Hz, 1H), 3.51(m, 2H), 2.05 (m, 1H), 1.87 (m, 1H), 1.66 (m, 4H).
Compound IIA-23: yellow solid, the yield was 87.1. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.31-8.20 (m, 2H), 7.98 (s, 1H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), 7.23 (dtd, J=47.4, 14.8, 3.2 Hz, 2H), 4.89 (p, J=5.6 Hz, 1H), 4.55 (t, J=9.0 Hz, 1H), 3.66 (qd, J=24.8, 5.4 Hz, 2H), 2.21 (ddd, J=22.9, 8.0, 4.6 Hz, 1H), 2.14 (ddd, J=14.7, 8.0, 4.6 Hz, 1H).
Compound IIA-24: yellow solid, the yield was 83.0%. 1H NMR (400 MHz, DMSO-d6) δ12.52 (d, J=3.3 Hz, 1H), 9.28 (d, J=3.2 Hz, 1H), 8.70 (s, 1H), 8.57 (d, J=8.1 Hz, 1H), 8.42-8.28 (m, 1H), 7.63-7.52 (m, 1H), 7.31 (dd, J=6.4, 2.9 Hz, 2H), 4.58 (dt, J=8.5, 4.3 Hz, 1H), 3.92 (qd, J=11.3, 4.5 Hz, 2H).
Compound IIA-25: yellow solid, the yield was 87.2%. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.27 (dd, J=14.1, 3.9 Hz, 2H), 7.98 (s, 1H), 7.54 (dd, J=14.6, 3.4 Hz, 1H), 7.28 (td, J=14.9, 3.4 Hz, 1H), 7.19 (td, J=14.8, 3.2 Hz, 1H), 4.20 (t, J=15.9 Hz, 1H), 3.54 (m, 5H), 2.23 (m, 1H), 1.75(m, 5H).
Compound IIA-26: yellow solid, the yield was 89.1%. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.27 (dd, J=14.7, 3.4 Hz, 1H), 8.18 (s, 1H), 7.98 (s, 1H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), 7.23 (dtd, J=47.4, 14.8, 3.2 Hz, 2H), 4.99 (t, J=8.6 Hz, 1H), 4.77 (p,J=16.0 Hz, 1H), 4.11 (dd, J=24.9, 15.7 Hz, 1H), 3.66 (s, 3H), 3.24 (dd, J=24.7, 15.8 Hz, 1H), 2.60 (ddd, J=24.9, 16.2, 8.6 Hz, 1H), 1.81 (ddd, J=24.7, 16.2, 8.7 Hz, 1H), 1.38 (s, 1H).
Compound IIA-27: yellow solid, the yield was 82.2%. 1H NMR (400 MHz, DMSO-d6) δ12.42 (s, 1H), 9.43 (d, J=1.2 Hz, 1H), 9.31 (t, J=6.3 Hz, 1H), 8.75 (s, 1H), 8.35 (d, J=6.9 Hz, 1H), 7.59 (dd, J=5.8, 2.7 Hz, 1H), 7.37-7.25 (m, 2H), 4.17 (dt, J=18.7, 9.4 Hz, 2H).
Compound IIA-28: yellow solid, the yield was 82.6%. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.27 (dd, J=14.7, 3.4 Hz, 1H), 8.19 (s, 1H), 8.07-7.96 (m, 2H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), 7.23 (dtd, J=47.4, 14.8, 3.2 Hz, 2H), 7.09 (dd, J=16.0, 10.0 Hz, 1H), 4.88 (t, J=8.6 Hz, 1H), 3.86 (dt, J=24.8, 13.2 Hz, 1H), 3.64 (dt, J=24.8, 13.1 Hz, 1H), 2.33-2.17 (m, 1H), 2.08-1.93 (m, 1H), 1.86-1.72 (m, 2H).
Compound IIA-29: yellow solid, the yield was 85.9%. 1H NMR (400 MHz, CDCl3) δ8.77-8.72 (m, 2H), 8.69 (dd, J=4.1, 1.7 Hz, 1H), 8.11 (s, 1H), 6.98 (dd, J=7.0, 4.1 Hz, 1H), 3.86-3.66 (m, 2H), 2.51 (qt, J=10.8, 6.8 Hz, 2H).
Compound IIA-30: yellow solid, the yield was 82.0%. 1H NMR (400 MHz, CDCl3) δ9.27 (s, 1H), 8.88 (s, 1H), 8.50 (d, J=7.3 Hz, 1H), 8.37 (s, 1H), 7.46 (d, J=7.6 Hz, 1H), 7.39-7.29 (m, 3H), 7.18 (s, 1H), 5.32 (t, J=6.2 Hz, 1H), 5.08 (t, J=6.0 Hz, 1H), 4.12 (t, J=5.6 Hz, 2H), 2.13-2.01 (m, 4H), 1.74 (s, 3H), 1.66 (s, 3H), 1.59 (s, 3H).
Starting from various substituted diazonium compounds 20, compound IIB was synthesized using a method similar to the one described above in this example. The structures of diazonium compound 20 and the resulting compound IIB are shown in Table 4:
Compound IIB-1: yellow solid, the yield was 43.1%. 1H NMR (400 MHz, CDCl3) δ9.74 (s, 1H), 8.26 (dd, J=14.7, 3.4 Hz, 1H), 8.20 (s, 1H), 7.97 (s, 1H), 7.53 (dd, J=14.6, 3.4 Hz, 1H), 7.48 (s, 1H), 7.22 (m, 7H), 5.62 (s, 2H), 4.36 (dt, J=55.1, 11.2 Hz, 2H), 3.06 (t, J=11.2 Hz, 2H).
Compound IIB-2: yellow solid, the yield was 41.0%. 1H NMR (400 MHz, DMSO-d6) δ12.38 (d, J=3.3 Hz, 1H), 9.07 (d, J=3.2 Hz, 1H), 8.88 (s, 1H), 8.41 (s, 1H), 8.37-8.26 (m, 1H), 7.98-7.86 (m, 4H), 7.67-7.56 (m, 1H), 7.51 (ddd, J=17.4, 7.3, 2.5 Hz, 3H), 7.36-7.24 (m, 2H), 5.81 (s, 2H), 5.48 (s, 2H).
Compound IIB-3: yellow solid, the yield was 49.3%. 1H NMR (400 MHz, CDCl3) δ9.75 (s, 1H), 8.27 (q, J=3.8 Hz, 2H), 7.98 (s, 1H), 7.54 (dd, J=14.7, 3.4 Hz, 1H), 7.23(m, 3H), 5.63 (s, 2H), 4.46 (t, J=15.4 Hz, 2H), 3.52 (t, J=15.6 Hz, 2H), 1.86(m, 4H), 1.33 (m, 4H).
Compound IIB-4: yellow solid, the yield was 50.0%. 1H NMR (400 MHz, CDCl3) δ9.75 (s, 1H), 8.36 (s, 1H), 8.27 (dd, J=14.7, 3.3 Hz, 1H), 7.98 (s, 1H), 7.54 (dd, J=14.6, 3.4 Hz, 1H), 7.39 (s, 1H), 7.23 (dtd, J=47.6, 14.9, 3.3 Hz, 2H), 5.63 (s, 2H), 4.46 (t, J=10.8 Hz, 2H), 3.62 (t, J=9.9 Hz, 2H), 1.95 (ddt, J=25.8, 15.0, 10.8 Hz, 2H), 1.77 (s, 1H), 1.46 (m, 6H).
Compound IIB-5: yellow solid, the yield was 59.3%. 1H NMR (400 MHz, DMSO-d6) δ12.38 (s, 1H), 9.08 (d, J=2.1 Hz, 1H), 8.88 (s, 1H), 8.39-8.27 (m, 1H), 8.16 (s, 1H), 7.63-7.56 (m, 1H), 7.31 (qd, J=7.1, 3.7 Hz, 2H), 5.49 (s, 2H), 4.08 (s, 2H), 1.99-1.89 (m, 3H), 1.65 (d, J=12.2 Hz, 3H), 1.53 (d, J=12.1 Hz, 3H), 1.46 (d, J=2.8 Hz, 6H).
Lymphocytotoxicity test: rats were euthanized by spinal dislocation method, their spleens were aseptically removed and ground to make a single cell suspension. After removing red blood cells with 2 mL of red blood cell lysis buffer, the cell concentration was adjusted to 2×106 cells/mL using RPMI-1640 medium containing 10% fetal bovine serum (FBS). Next, 100 μL of the 2×106 cells/mL cell suspension and 100 μL of the appropriate concentration of the tested compound were added to each well of a 96-well plate, and the plate was incubated at 37° C. with 5% CO2 for 48 hours. At the end of the incubation period, 10 μL of CCK8 was added to each well, and the plate was returned to the incubator for 5-7 hours. The OD450 value was then measured at 450 nm using an enzyme-linked immunosorbent assay reader.
Lymphocyte proliferation assay: fresh spleen cells at a concentration of 2×106 cells/mL were incubated for 48 hours in an incubator maintained at 37° C. and 5% CO2 for 48 hours, and the culture medium was induced for cell proliferation with 5 μg/mL of ConA, and the appropriate concentration of the tested compound was added to test its inhibitory activity on lymphocyte proliferation. The test results of some compounds are shown in the table below and figures accompanying the application:
Wherein Tacrolimus is positive drug Tacrolimus; ITE is an endogenous AhR partial agonist (natural endogenous ligand of AHR); KYN (kynurenine) is metabolized by Trp via the dioxygenases TDO and IDO and is an AhR agonist.
The compounds obtained in the example of the present application have better inhibitory activity on lymphocyte proliferation, wherein IC50 values of some compounds are shown in the following table, wherein the IC50 curves of some compounds are shown in
In addition, the toxicity of the compounds on lymphocytes was also tested in this example. The testing method was as follows: the cell density was 3×106 cells/mL, and 100 μL of the cell suspension containing 3×105 cells/well was added to each well of a 96-well plate without any stimulation factor. The toxicity of the compounds was then tested at concentrations of 5, 2.5, 1.25, and 0.625 The test results showed that the compound in the present application had no obvious toxicity at the test concentration, and the results were shown in
The results showed that the compounds of the present application have low toxicity to lymphocytes and have a significant inhibitory effect on lymphocytes proliferation. Among them, IA-3 has a particularly significant inhibitory effect on lymphocytes and has less cytotoxicity and high safety index. The remaining compounds also showed significant inhibitory activity on the proliferation of rat lymphocytes and were generally superior to the positive drug Tacrolimus.
In conclusion, it can be concluded from these facts that the compounds of the present application are effective in inhibiting immune cell activity, can be used for the prevention or treatment of immune diseases, and have very promising applications in the preparation of immunosuppressive drugs.
A total of 15 healthy male mice were randomly and equally divided into 3 groups, namely, control group, model group and drug administration group, with 5 mice in each group, labeled and their body weight recorded. Respectively administered: the control group and the model group were injected with normal saline via tail vein respectively, and the administration group was injected with the prepared in Examples 1-4 via tail vein respectively; the dose was 10 mg/kg (2 mg/mL; 50 μL/10 g). One hour later, LPS was injected intraperitoneally at 3 mg/kg (0.3 mg/mL, 100 μL/10 g). A second injection of the drug was given 5 hours later. Blood was collected from the mice 9 hours later, followed by euthanasia.
ELISA was performed to determine levels of IL-6, IL-2, TNF-a and IFN-y in serum. The results showed that the compounds of the present application exhibited different degrees of reduction in the levels of LPS-induced inflammatory factors IL-6, IL-2, TNF-a and IFN-y in serum, especially compound IA-3 had the most significant effect on the reduction of the above factors, and the results are shown in
Histopathological examination was performed on the administration group of compound IA-3: mouse lungs were fixed with 4% paraformaldehyde, and histopathological changes were observed by HE. The results showed that the LPS group had severe lung injury, manifested by inflammatory cell infiltration, thickening of alveolar walls, pulmonary interstitial congestion, and bleeding. However, treatment with compound IA-3 reduced the severity of lung pathology. The results of H&E staining of mouse lung tissues were shown in
A total of 15 healthy male mice were randomly and equally divided into 3 groups, namely, control group, model group and drug administration group, with 5 mice in each group, labeled and their body weight recorded. Respectively administered: the control group and the model group were injected with normal saline via tail vein respectively, and the administration group was injected with the compounds prepared in Examples 1-4 via tail vein respectively; the dose was 10 mg/kg (2 mg/mL; 50 μL/10 g). CD-3 antibody was injected intraperitoneally, 100 μg/kg, after 0.5 hours. A second injection of the drug was given 5 hours later. Blood was collected from the mice 9 hours later, followed by euthanasia.
ELISA was performed to determine levels of IL-6, IL-2, TNF-a and IFN-y in serum. The results showed that the compounds of the present application exhibited different degrees of reduction in the levels of CD-3 antibody-induced inflammatory factors IL-6, IL-2, TNF-a and IFN-y in serum, especially compound IA-3 had the most significant effect on the reduction of the above factors, and the results are shown in
After experimental verification that the compounds of the present application have good oral bioavailability, this example uses compound IA-3, which exhibits relatively good inhibitory activity on immune cell proliferation, as an example, to illustrate the process of studying its oral bioavailability prior to its further study as an immunosuppressant in animal models, as follows:
To prepare the standard series solution, a suitable amount of compound IA-3 was accurately weighed and placed in a 100 mL volumetric flask. The compound was dissolved in acetonitrile and diluted to the mark to produce a 25.0 μg/mL stock solution. The solution was mixed thoroughly by shaking and used for further analysis. An appropriate amount of the stock solution was taken separately, diluted with acetonitrile, and made up to the mark to prepare a series of standard solutions with concentrations of 5, 10, 25, 50, 100, 250, 500, and 1000 ng/mL. The solutions were stored in a refrigerator at 4° C. for later use.
Preparation of internal standard solution: a suitable amount of internal standard was accurately weighed and placed in a 100 mL volumetric flask. The internal standard was dissolved in acetonitrile and diluted to the mark to produce a 25 μg/mL stock solution. The solution was mixed thoroughly by shaking and used for further analysis. An accurately weighed amount of the stock solution was taken again and diluted with acetonitrile to prepare the internal standard reference solution at a concentration of 50 ng/mL. The solution was stored in a refrigerator at 4° C. for later use.
Preparation of plasma samples: 80 μL of rat plasma was taken, and 120 μL of internal standard was added. The mixture was vortexed for 2 min and centrifuged for 10 min. Then, 140 μL of the supernatant was collected and centrifuged for another 10 min. Finally, 10 μL of the supernatant was injected for analysis.
Standard Curve: 64 μL of blank rat plasma was mixed with 16 μL of the test compound standard series solution and 120 μL of acetonitrile solution containing the internal standard. The mixture was vortexed for 2 minutes and centrifuged for 10 minutes. Then, 140 μL of the supernatant was taken and centrifuged for another 10 minutes, 10 μL of the sample was injected and analyzed with HPLC-HR-MS. Instrumentation obtained the linear regression equation as the standard curve.
Sampling: eight healthy male Wistar rats were randomly divided into two groups (oral group and intravenous group), with four rats in each group. Rats were fasted for 12 h and provided with free access to water before the experiment. The oral group was administered with the test drug at a dose of 5 mg/kg, and the intravenous group was administered with 1 mg/kg. Blood samples of approximately 0.4 mL were collected from the rat orbital vein before and at 5 min, 15 min, 30 min, 1 h, 1.5 h, 2 h, 4 h, 6 h, 8 h, 10 h, and 12 h after administration, and immediately transferred to heparinized centrifuge tubes. Plasma was separated by centrifugation.
Sample determination and data processing: the rat plasma samples were processed according to the “Preparation of plasma samples” procedure and analyzed under the conditions described above. The non-compartmental model was applied to process the drug concentration data mentioned above, and the pharmacokinetic parameters were calculated using the DAD 2.0 software. The bioavailability was calculated according to the formula F=AUCig*Div/AUCiv*Dig*100%.
The results were shown in Tables 5 and 6:
After intragastric administration of IA-3, the total exposure (AUC) in whole blood was higher, with a relative oral bioavailability of 22.07%. These results suggest that IA-3 has the potential for becoming a drug.
The aforementioned descriptions are only preferred embodiments of the present application and are not intended to limit the scope of the present application. Although the aforementioned embodiments have been described in detail, those skilled in the art can still modify the technical solutions described in the embodiments or replace some technical features with equivalent alternatives. Any modifications, equivalent replacements, improvements, and the like made within the spirit and principles of the present application should be included within the scope of the present application's protection.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2021105834508 | May 2021 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2021/098707 | 6/7/2021 | WO |
