WO2024179405A1 - Class of cinnamamide derivatives and application thereof - Google Patents

Abstract

The present invention relates to a class of cinnamamide derivatives and an application thereof. Specifically, compounds of the present invention have a structure shown in Formula I, wherein the definitions of groups and substituents are as described in the description. Further disclosed in the present invention are a preparation method of the compounds and a use thereof in preventing and/or treating GPR183-related diseases.

Description

A class of cinnamamide derivatives and their application Technical Field

The invention relates to the field of medicine, and in particular to a class of cinnamamide derivatives and applications thereof.

Background Art

G protein-coupled receptor 183 (GPR183), also known as Epstein-Barr virus-induced gene 2 (EBI2), was first identified in 1993 as a lymphocyte-specific GPCR expressed at high levels in EBV-infected cells. In 2011, hydroxycholesterols (oxysterols) were identified as endogenous agonists of GPR183, with 7α,25-dihydroxycholesterol (7α,25-OHC) being the most potent endogenous ligand. Subsequently, GPR183 was identified as a chemotactic receptor, with 7α,25-dihydroxycholesterol being a potent endogenous ligand, a finding that correlated well with the observation that GPR183 is highly expressed in leukocytes and that its differential expression is important for accurate localization of B cells within lymphoid organs. The pharmacological significance of this target stems from the role of GPR183 itself and its endogenous ligands in a variety of diseases, such as B cell malignancies, inflammatory/autoimmune diseases, and metabolic diseases. The activation of GPR183 plays a vital role in humoral immune response. Many autoimmune diseases are directly or indirectly related to the physiological function of GPR183. GPR183 agonists and antagonists are of great value in further explaining its biological function and its role in the treatment of various diseases. In 2014, through high-throughput screening and structural optimization, the small molecule compound NIBR189 was identified as the first GPR183 antagonist, showing strong inhibitory activity against agonist-induced GPR183 activation.

Summary of the invention

The object of the present invention is to provide a compound represented by formula I and a preparation method thereof and use thereof in preventing and/or treating GPR183-related diseases.

The first aspect of the present invention provides a compound, which is a compound represented by formula I, or A pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof,

Wherein, ring A is a saturated or unsaturated 4-10 membered heterocycloalkyl containing 1, 2 or 3 N;

n is selected from the following group: 0, 1, 2, 3;

Each R ₁ , R ₂ , and R ₃ is independently selected from the following group: hydrogen, deuterium, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C4 alkyl-(C＝O)-NH-, -N(C1-C6 alkyl) ₂ , -NH(C1-C6 alkyl), -NH ₂ , wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: halogen, hydroxyl, amino;

Y is a straight bond or a C1-C5 alkylene group;

When Y is a straight bond, T is R ₄ and R ₅ are each independently selected from the following group: H, C1-C6 alkoxy; or, R ₄ and R ₅ together with the phenyl carbon to which they are connected form a substituted or unsubstituted 3-6 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 3-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, halogen, =O, hydroxyl, mercapto, amino, -NH(C1-C6 alkyl), -N(C1-C6 alkyl) ₂ ;

When Y is C1-C5 alkylene, T is R ₄ and R ₅ are each independently selected from the following group: H, C1-C6 alkoxy; or, R ₄ and R ₅ together with the phenyl carbon to which they are connected form a substituted or unsubstituted 3-6 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 3-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, halogen, =O, hydroxyl, mercapto, amino, -NH(C1-C6 alkyl), -N(C1-C6 alkyl) ₂ ;

R' is selected from the following group: hydrogen, deuterium, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C4 alkyl-(C＝O)-NH-, wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: halogen, hydroxyl, amino;

m is selected from the following group: 1, 2, 3;

The additional condition is that the compound is not

In another preferred embodiment, _R4 and _R5 together with the phenyl carbon to which they are connected form a substituted or unsubstituted 3-6 membered heteroaryl group containing 1, 2 or 3 heteroatoms selected from N, O or S.

In another preferred embodiment, R ₄ and R ₅ together with the phenyl carbon to which they are connected form a substituted 5-membered heteroaryl containing 1 N and 1 S, wherein the substitution refers to substitution by a group selected from the group consisting of: -N(C1-C6 alkyl) ₂ , -NH(C1-C6 alkyl), -NH ₂ .

In another preferred embodiment, the 5-membered heteroaryl group containing 1 N and 1 S is

In another preferred embodiment, Select from the following group:

In another preferred embodiment, ring A is a ring structure selected from the following group: a monocyclic ring, a fused ring, a bridged ring, and a spirocyclic ring.

In another preferred embodiment, ring A is a saturated 4-8 membered heterocycloalkyl group containing 2 N atoms, and ring A is a monocyclic ring;

n is 0;

Each R ₁ , R ₂ , and R ₃ is independently selected from the group consisting of hydrogen, halogen;

Y is a straight bond, T is for

X ₁ , X ₂ , and X ₃ are each independently selected from the group consisting of N, CR ₆ , C(R ₆ ) ₂ , NR ₆ , S, and O;

Each R ₆ is independently selected from the group consisting of hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, halogen, =0, hydroxyl, mercapto, and amino.

In another preferred embodiment, It means that the structure is unsaturated or aromatic.

In another preferred embodiment, ring A is

In another preferred embodiment, Select from the following group:

In another preferred embodiment, ring A is a saturated 6-10 membered heterocycloalkyl group containing 2 N atoms, and ring A is a bridged ring;

n is 0;

Each R ₁ , R ₂ , and R ₃ is independently selected from the group consisting of hydrogen, halogen;

Y is a straight bond, T is _R4 and _R5 are each independently selected from the group consisting of H, C1-C6 alkoxy.

In another preferred embodiment, ring A is

In another preferred embodiment, ring A is a saturated 4-8 membered heterocycloalkyl group containing 1 N, and ring A is a monocyclic ring;

n is selected from the following group: 0, 1, 2, 3;

Each R ₁ , R ₂ , and R ₃ is independently selected from the group consisting of hydrogen, halogen;

Y is C1-C5 alkylene, T is _R4 and _R5 are each independently selected from the following group: H, C1-C6 alkoxy;

R' is selected from the group consisting of hydrogen, halogen;

m is selected from the following group: 1, 2, 3.

In another preferred embodiment, ring A is

In another preferred embodiment, n is 0, Y is C1-C3 alkylene, and T is

R' is a halogen;

m is selected from the following group: 1, 2, 3.

In another preferred embodiment, ring A is a saturated 4-8 membered heterocycloalkyl group containing 2 N atoms, and ring A is a monocyclic ring;

n is 0;

Each of R ₁ , R ₂ , and R ₃ is independently halogen;

Y is a straight bond, T is for

X ₁ , X ₂ , and X ₃ are each independently selected from the following group: N, CR ₆ , C(R ₆ ) ₂ , NR ₆ , S, and O;

Each R ₆ is independently selected from the group consisting of hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, halogen, =0, hydroxyl, mercapto, and amino.

In another preferred embodiment, ring A is

In another preferred embodiment, Select from the following group:

In another preferred embodiment, the compound is selected from the following group:

The second aspect of the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a safe and effective amount of one or more compounds described in the first aspect of the present invention.

The third aspect of the present invention provides a use of the compound described in the first aspect of the present invention for preparing a medicament for preventing and/or treating a GPR183-related disease.

In another preferred embodiment, the drug is used to inhibit the activity of GPR183.

In another preferred embodiment, the GPR183-related disease is selected from the following group: tumor, inflammation, autoimmune disease, metabolic disease, pain, bacterial infection disease, viral infection disease.

In another preferred embodiment, the tumor is selected from the following group: B-cell malignancies, hematological malignancies, clear cell renal cell carcinoma, prostate cancer, laryngeal squamous cell carcinoma, head and neck cancer, and non-small cell lung cancer.

In another preferred embodiment, the inflammation is selected from the group consisting of inflammatory bowel disease, osteoarthritis, non-alcoholic fatty liver disease, interstitial cystitis, atherosclerosis, pneumonia, chronic sinusitis, myocarditis, and nephritis.

In another preferred embodiment, the autoimmune disease is selected from the following group: rheumatoid arthritis, systemic lupus erythematosus, autoimmune encephalomyelitis, vitiligo, type Ⅰ diabetes, chronic atrophic gastritis, multiple sclerosis, acute idiopathic polyneuritis.

In another preferred embodiment, the metabolic disease is selected from the group consisting of obesity, diabetes, dyslipidemia, osteoporosis, and scurvy.

In another preferred embodiment, the pain is neuropathic pain.

In another preferred embodiment, the bacterial infection disease is selected from the following group: pulmonary tuberculosis, scarlet fever, and purulent meningitis.

In another preferred embodiment, the viral infectious disease is selected from the following group: new coronavirus, influenza virus, measles, mumps, and viral hepatitis.

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 shows the inhibitory activity results of representative compounds S10 and NIBR189 obtained in Experimental Example 3 on GPR183, (a) the activity test results of S10 under the Glosensor cAMP test system; (b) the activity test results of S10 under the CRE reporter gene test system; (c) the activity test results of NIBR189 under the Glosensor cAMP test system; (d) the activity test results of NIBR189 under the CRE reporter gene test system.

FIG. 2 shows the inhibitory effect of the representative compound S10 obtained in Experimental Example 4 on hERG potassium ion channels.

DETAILED DESCRIPTION

After long-term and in-depth research, the inventor unexpectedly prepared a compound of formula I with stronger GPR183 inhibitory activity and better pharmacokinetic properties through structural optimization. On this basis, the inventor completed the present invention.

the term

In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

In the present invention, the term "halogen" refers to F, Cl, Br or I.

In the present invention, "C1-C6 alkyl" refers to a straight or branched alkyl group comprising 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl, tert-pentyl, or the like.

In the present invention, the term "C2-C6 alkenyl" refers to a straight or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl.

In the present invention, the term "C2-C6 alkynyl" refers to a straight or branched alkynyl group having 2 to 6 carbon atoms and containing one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl.

In the present invention, the term "C3-C8 cycloalkyl" refers to a cyclic alkyl group having 3 to 8 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy and butoxy, etc. Preferably, it is C1-C4 alkoxy.

In the present invention, the term "heterocyclyl" or "heterocycloalkyl" refers to a 3-10 membered heterocyclic group containing 1, 2 or 3 heteroatoms selected from N, O and S, including (but not limited to) the following groups:

In the present invention, the term "aromatic ring" or "aryl group" has the same meaning, preferably "C6-C10 aryl group". The term "C6-C10 aryl group" refers to an aromatic ring group having 6 to 10 carbon atoms without heteroatoms in the ring, such as phenyl, naphthyl, etc.

In the present invention, the term "aromatic heterocycle" or "heteroaryl" has the same meaning and refers to a heteroaromatic group containing one to multiple heteroatoms. For example, "3-6 membered heteroaryl" refers to an aromatic heterocycle containing 1 to 3 heteroatoms selected from oxygen, sulfur and nitrogen and 1 to 5 carbon atoms. Non-limiting examples include: furanyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring. The heteroaryl group can be optionally substituted or unsubstituted.

In the present invention, the term "halo" means substituted with halogen.

In the present invention, the term "deuterated" means substituted with deuterium.

In the present invention, the term "substituted" refers to one or more hydrogen atoms on a specific group being replaced by a specific substituent. The specific substituent is the substituent described above, or the substituent appearing in each embodiment. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable position of the group, and the substituent may be the same or different at each position. It should be understood by those skilled in the art that the combination of substituents contemplated by the present invention is those stable or chemically feasible combinations. The substituents include, for example (but not limited to): halogen, hydroxyl, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3- to 12-membered heterocyclic radical, aryl, heteroaryl, C1-C8 aldehyde, C2-C10 acyl, C2-C10 ester, amino, C1-C6 alkoxy, C1-C10 sulfonyl, etc.

In the present invention, the term 1-6 refers to 1, 2, 3, 4, 5 or 6. Other similar terms independently have similar meanings. The term "plurality" refers to 2-6, such as 2, 3, 4, 5 or 6.

It should be understood that when a group is present in multiple different positions of a compound at the same time, its definition at each position is independent of each other and may be the same or different. That is, the term "selected from the following group:" has the same meaning as the term "each independently selected from the following group:".

Compound

The present invention designs and synthesizes a new type of cinnamoyl derivatives through the drug chemistry techniques such as ring-integration strategy and fluorine substitution. The compounds show better inhibitory activity than NIBR189 and better pharmacokinetic properties. Has potential for further development.

Specifically, the present invention provides a compound, which is a compound represented by Formula I, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof,

wherein each group is as defined above.

In another preferred embodiment, in the compound, any one of ring A, R ₁ , R ₂ , R ₃ , n, m, R ₄ , R ₅ , Y, T, and R′ is independently a corresponding group in the specific compound of the present invention.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a compound of the present invention and an acid or base that is suitable for use as a drug. Pharmaceutically acceptable salts include inorganic salts and organic salts. A preferred class of salts is a salt formed by a compound of the present invention and an acid. Suitable acids for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid.

Another preferred salt is a salt of the compound of the present invention and a base, such as an alkali metal salt (e.g., sodium salt or potassium salt), an alkaline earth metal salt (e.g., magnesium salt or calcium salt), an ammonium salt (e.g., lower alkanolammonium salt and other pharmaceutically acceptable amine salts), for example, methylamine salt, ethylamine salt, propylamine salt, dimethylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tert-butylamine salt, ethylenediamine salt, hydroxyethylamine salt, dihydroxyethylamine salt, trihydroxyethylamine salt, and amine salts formed from morpholine, piperazine, and lysine, respectively.

The term "solvate" refers to a complex in which the compound of the present invention is coordinated with solvent molecules to form a specific ratio.

It should be understood that the embodiments of the present invention specifically describe the preparation methods of the compounds of formula I of the present invention, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention belongs.

Typically, the raw materials and reagents used in the preparation process of the compounds of the present invention can be purchased through commercial channels unless otherwise specified.

Pharmaceutical compositions and methods of administration

The present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and one or more safe and effective amounts of the compounds of the present invention.

Since the compounds of the present invention have excellent anti-tumor activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate tumor-related diseases.

The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-1000 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

The pharmaceutical composition is in the form of injection, capsule, tablet, pill, powder or granule.

There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic; (c) humectants, such as glycerol; (d) disintegrants. (e) dissolving agents, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) buffering agents, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain a buffer.

Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as anti-tumor drugs).

The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

Compared with the prior art, the present invention has the following main advantages:

(1) Compared with NIBR189, the compounds of the present invention have better GPR183 inhibitory activity and better pharmacokinetic properties;

(2) The compounds of the present invention can effectively inhibit the migration and activation of T cells.

The present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are generally carried out according to conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

Preparation Example 1 Preparation of Compound S1:

Synthesis of compound 1-2

Compound 1-1 (10.0 g, 44.05 mmol), 1-tert-butyloxycarbonyl-piperazine (8.2 g, 44.05 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (55.7 g, 147 mmol) were added. Dissolve in 80 ml of anhydrous N,N-dimethylformamide. Take triethylamine (37 ml, 267 mmol) and slowly add it to the stirring solution at room temperature. Stir at room temperature after the addition is complete. After reacting for 6 hours, the reaction solution is extracted with ethyl acetate, and the organic phase is washed with saturated brine and then dried over anhydrous sodium sulfate. Collect the organic phase, concentrate it, and then elute it with column chromatography gradient to obtain a pink powder solid 1-2.

Synthesis of Compounds 1-3

Compound 1-2 (4.0 g, 10.12 mmol) was dissolved in 45 ml of hydrochloric acid 1,4-dioxane solution and stirred at room temperature. After reacting for two hours, the reaction solution was filtered, the filter cake was washed with 1,4-dioxane solution and placed in a drying oven for drying, and finally an off-white powder solid 1-3 was obtained.

Synthesis of compound S1

Compound 1-3 (80 mg, 0.27 mmol), 1-methyl-1H-indole-6-carboxylic acid (47 mg, 0.27 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (124 mg, 0.33 mmol) were dissolved in 3 ml of anhydrous N,N-dimethylformamide. N-methylmorpholine (150 μl, 1.35 mmol) was slowly added dropwise to the stirring solution, and stirred at room temperature after the addition was completed. After reacting for 3 hours, the reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine and then dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, and then gradient eluted by column chromatography to obtain a white powder solid S1. ¹ H NMR(400MHz,chloroform-d)δ7.74(s,1H),7.64(d,J＝15.4Hz,1H),7.51(d,J＝8.2Hz,2H),7.39(d,J＝8.2Hz,2H),7.36–7.29(m,2H),7.13(d,J＝3.1Hz,1H),6.86 (d,J＝15.4Hz,1H),6.54(d,J＝3.1Hz,1H),3.83(s,3H),3.82–3.46(m,8H).

Preparation Example 2 Preparation of Compound S2:

Substrate 1-4 was replaced with 2-methylbenzimidazole-5-carboxylic acid, and the synthesis steps of compound S2 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ7.64 (d, J = 15.4 Hz, 1H), 7.51 (d, J = 8.1 Hz, 3H), 7.39 (d, J = 8.0 Hz, 2H), 7.28 (s, 3H), 6.86 (d, J = 15.4 Hz, 1H), 3.77 (s, 3H), 3.88-3.57 (m, 8H).

Preparation Example 3 Preparation of Compound S3:

Substrate 1-4 was replaced with 2-mercapto-5-benzimidazolecarboxylic acid, and the synthesis steps of compound S3 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ7.62 (d, J = 15.6 Hz, 1H), 7.53 (d, J = 7.9 Hz, 2H), 7.42 (d, J = 8.2 Hz, 2H), 7.36–7.31 (m, 4H), 7.31–7.27 (m, 1H), 6.89 (d, J = 15.1 Hz, 1H), 3.88–3.32 (m, 8H).

Preparation Example 4 Preparation of Compound S4:

Substrate 1-4 was replaced with benzothiazole-6-carboxylic acid, and the synthesis steps of compound S4 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ9.11 (s, 1H), 8.19 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 1.7 Hz, 1H), 7.65 (d, J = 15.4 Hz, 1H), 7.57 (dd, J = 8.4, 1.6 Hz, 1H), 7.51 (d, J = 8.1 Hz, 2H), 7.39 (d, J = 8.1 Hz, 2H), 6.86 (d, J = 15.3 Hz, 1H), 3.96–3.46 (m, 8H).

Preparation Example 5 Preparation of Compound S5:

Substrate 1-4 was replaced with 2,2-difluorobenzodioxolane-5-carboxylic acid, and the synthesis steps of compound S5 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ7.65 (d, J = 15.3 Hz, 1H), 7.54–7.48 (m, 2H), 7.39 (d, J = 8.1 Hz, 2H), 7.19 (tt, J = 5.0, 1.5 Hz, 2H), 7.12 (dd, J = 8.6, 1.0 Hz, 1H), 6.85 (d, J = 15.4 Hz, 1H), 3.73 (m, 8H).

Preparation Example 6 Preparation of Compound S6:

Substrate 1-4 was replaced with 2-oxo-2,3-dihydro-1H-benzo[d]imidazole-5-carboxylic acid, and the synthesis steps of compound S6 were the same as those of compound S1. ¹ H NMR (400 MHz, DMSO-d ₆ )δ10.83(s,1H),10.78(s,1H),7.69(s,2H),7.61(s,2H),7.49(d,J＝15.2Hz,1H),7.32(d,J＝15.7Hz,1H),7.04(s,1H),6.98(s,2H),3.81–3.45(m,8H).

Preparation Example 7 Preparation of Compound S7:

Substrate 1-4 was replaced with 3-methyl-1H-indole-6-carboxylic acid, and the synthesis steps of compound S7 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ8.28 (s, 1H), 7.70–7.55 (m, 2H), 7.54–7.47 (m, 3H), 7.39 (d, J = 8.1 Hz, 2H), 7.16 (dd, J = 8.1, 1.4 Hz, 1H), 7.11–7.04 (m, 1H), 6.86 (d, J = 15.4 Hz, 1H), 4.11-3.58 (m, 8H), 2.34 (d, J = 1.1 Hz, 3H).

Preparation Example 8 Preparation of Compound S8:

Substrate 1-4 was replaced with 2-methyl-1H-indole-6-carboxylic acid, and the synthesis steps of compound S8 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ8.35 (s, 1H), 7.64 (d, J = 15.4 Hz, 1H), 7.55–7.48 (m, 3H), 7.45 (s, 1H), 7.39 (d, J = 8.2 Hz, 2H), 7.11 (dd, J = 8.1, 1.5 Hz, 1H), 6.86 (d, J = 15.5 Hz, 1H), 6.25 (s, 1H), 3.99–3.56 (m, 8H), 2.47 (s, 3H).

Preparation Example 9 Preparation of Compound S9:

Substrate 1-4 was replaced with 1H-indole-6-carboxylic acid, and the synthesis steps of compound S9 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ8.63 (s, 1H), 7.71–7.60 (m, 2H), 7.56 (s, 1H), 7.51 (d, J = 8.2 Hz, 2H), 7.39 (d, J = 8.1 Hz, 2H), 7.32 (t, J = 2.9 Hz, 1H), 7.16 (dd, J＝8.2,1.4Hz,1H),6.86(d,J＝15.3Hz,1H),6.59(d,J＝2.8Hz,1H),3.73(m,8H).

Preparation Example 10 Preparation of Compound S10:

Substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid, and the synthesis steps of compound S10 were the same as those of compound S1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.79 (d, J=1.7 Hz, 1H), 7.69 (d, J=8.0 Hz, 4H), 7.61 (d, J=8.3 Hz, 2H), 7.49 (d, J=15.4 Hz, 1H), 7.39–7.26 (m, 3H), 3.83–3.44 (m, 8H).

Preparation Example 11 Preparation of Compound S11:

Substrate 1-4 was replaced with 2-trifluoromethylbenzothiazole-6-carboxylic acid, and the synthesis steps of compound S11 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ8.27 (d, J = 8.4 Hz, 1H), 8.13 (s, 1H), 7.67 (d, J = 2.2 Hz, 1H), 7.64 (d, J = 5.4 Hz, 1H), 7.52 (d, J = 8.0 Hz, 2H), 7.39 (d, J = 8.1 Hz, 2H), 6.85 (d, J = 15.3 Hz, 1H), 4.04–3.36 (m, 8H).

Preparation Example 12 Preparation of Compound S12:

Synthesis of compound 12-2

Compound 1-1 (150 mg, 0.66 mmol), compound 12-1 (140 mg, 0.66 mmol) and HATU (836 mg, 2.2 mmol) were dissolved in 3 ml of anhydrous N,N-dimethylformamide. Triethylamine (556 μl, 4 mmol) was slowly added to the stirring solution at room temperature, and stirred at room temperature after the addition was completed. After reacting for 6 hours, the reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine and then dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, and then subjected to column chromatography gradient elution to obtain a white powder solid 12-2.

Synthesis of compound 12-3

Compound 12-2 (111 mg, 0.264 mmol) was dissolved in 2 ml of dichloromethane, and 1 ml of trifluoroacetic acid was added. After reacting at room temperature for 2 hours, the mixture was concentrated and saturated sodium bicarbonate solution was slowly added to neutralize the mixture. The mixture was extracted with methane, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography with gradient elution to obtain a white solid 12-3.

Synthesis of compound S12

Compound 12-3 (157 mg, 0.49 mmol) was dissolved in 3 ml of anhydrous dichloromethane and stirred in an ice bath. Triethylamine (613 μl, 4.42 mmol) was slowly added to the stirred solution in an ice bath. After stirring for two minutes, 4-methoxybenzoyl chloride (100 μl, 0.74 mmol) was slowly added dropwise. After reacting for half an hour, the solution was filtered, and the filter cake was washed with a small amount of anhydrous ethanol and dried to obtain a white solid S12. ¹ H NMR (400MHz, chloroform-d) δ7.69 (d, J = 15.4Hz, 1H), 7.52 (d, J = 8.0Hz, 2H), 7.40 (d, J = 7.9Hz, 2H), 7.36 (d, J = 8.3Hz, 2H), 6.92 (d, J = 8.2Hz, 2H), 6.76 (d, J = 15.5 Hz,1H),4.76(s,2H),4.46(s,1H),3.84(s,3H),3.52(s,1H),3.05(s,1H),2.04(s,2H),1.94(s,2H),1.65(s,1H).

Preparation Example 13 Preparation of Compound S13:

Substrate 12-1 was replaced with tert-butyl 2,5-diazabicyclo[2.2.2]octane-2-carboxylate, and the synthesis steps of compound S13 were the same as those of compound S12. ¹ H NMR (400 MHz, Methanol-d ₄ )δ7.61(d, J＝5.8 Hz, 1H),7.59–7.54(m, 4H),7.52(d, J＝10.9 Hz, 1H),7.48–7.38(m, 1H),7.24–6.95(m, 3H),4.81–3.92(m, 3H),3.84(s, 3H),3.83–3.57(m, 3H),2.08(s, 3H),1.88(s, 1H).

Preparation Example 14 Preparation of Compound S14:

Substrate 12-1 was replaced with tert-butyl (4-methylpiperidin-4-yl)carbamate, and the synthesis steps of compound S14 were the same as those of compound S12. ¹ H NMR (400MHz, chloroform-d) δ7.77(d,J＝8.3Hz,2H),7.59(d,J＝15.4Hz,1H),7.50(d,J＝7.9Hz,2H),7.37(d,J＝8.2Hz,2H),6.95(d,J＝8.2Hz,2H),6.86(d,J＝15.6 Hz,1H),6.37(s,1H),4.50(s,1H),3.94(s,1H),3.86(t,J＝1.5Hz,3H),3.76 (s,1H),3.68–3.40(m,2H),3.16(s,1H),1.97(d,J＝13.1Hz,2H),1.87–1.66 (m,2H).

Preparation Example 15 Preparation of Compound S15:

Synthesis of compound 15-3

Compound 15-1 (300 mg, 1.08 mmol), p-methoxyaniline (172 mg, 1.40 mmol) and [(dimethylamino)chloromethylene]dimethylammonium hexafluorophosphate (362 mg, 1.29 mmol) were dissolved in 3 ml of anhydrous acetonitrile and stirred at room temperature. Then 1-methylimidazole (309 mg, 3.76 mmol) was slowly added to the stirring solution. After reacting for 3 hours, the reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine and then dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, and then subjected to column chromatography gradient elution to obtain a red oily liquid 15-3.

Synthesis of compound 15-4

Compound 15-3 (188 mg, 0.49 mmol) was dissolved in 2 ml of methanol solution of hydrochloric acid and reacted at room temperature for 2 hours, then concentrated, slowly added with saturated sodium bicarbonate solution for neutralization, extracted with dichloromethane, the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to gradient elution by column chromatography to obtain dark purple solid 15-4.

Synthesis of compound S15

Compound 15-4 (139 mg, 0.49 mmol), compound 1-1 (133 mg, 0.59 mmol) and HATU (619 mg, 1.63 mmol) were dissolved in 3 ml of anhydrous N,N-dimethylformamide. Triethylamine (412 μl, 2.96 mmol) was slowly added to the stirring solution at room temperature, and the mixture was stirred at room temperature after the addition was completed. After reacting for 6 hours, the reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine and then dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, and then subjected to column chromatography gradient elution to obtain white powder solid S15. ¹ H NMR (400 MHz, DMSO-d ₆ )δ9.91(d,J＝4.5Hz,1H),7.73(t,J＝10.6Hz,2H),7.61(d,J＝8.0Hz,2H),7.53–7.46(m,3H),7.41–7.31(m,1H),6.88(d,J＝8.4Hz,2H),4.68(s,1H),4.38 (dd,J＝45.0,13.5Hz,1H),3.71(d,J＝2.3Hz,3H),3.67–3.48(m,1H),3.27(d,J＝6.8Hz,1H),2.67(d,J＝14.1Hz,2H),2.28(s,1H),1.88(s,1H),1.49–1.29 (m,1H).

Preparation Example 16 Preparation of Compound S16:

Substrate 15-1 was replaced with 2-(1-(tert-butyloxycarbonyl)piperidin-4-yl)-2,2-difluoroacetic acid, and the synthesis steps of compound S16 were the same as those of compound S15. ¹ H NMR(400MHz,chloroform-d)δ7.91(s,1H),7.58(d,J＝16.2Hz,1H),7.48(t,J＝9.2Hz,4H),7.37(d,J＝8.0Hz,2H),6.91–6.87(m,2H),6.84(s,1H),4.85(d,J＝14 .0Hz,1H),4.19(d,J＝11.5Hz,1H),3.80(d,J＝2.2Hz,3H),3.14(s,1H),2.81–2.44(m,2H),1.93(s,2H),1.52(s, 2H).

Preparation Example 17 Preparation of Compound S17:

Synthesis of compound 17-1

Fluorophosphonoacetate (257 mg, 1.20 mmol) was slowly added dropwise to a stirred suspension of sodium hydride (55% oil dispersion, 1.2 mmol) in 2 ml of anhydrous tetrahydrofuran under argon protection. After the addition was complete, the reaction solution was stirred at 0°C for 15 minutes. Subsequently, a solution of p-bromobenzaldehyde (185 mg, 1.00 mmol) in 2 ml of anhydrous tetrahydrofuran was slowly added dropwise to the stirred reaction solution and stirred for another 30 minutes at 0°C. The reaction was refluxed overnight and cooled to room temperature. The reaction was quenched with saturated aqueous ammonium chloride solution. The reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine and then dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, and then gradient eluted by column chromatography to obtain compound 17-1.

Synthesis of compound 17-2

Compound 17-1 (355 mg, 1.30 mmol) was dissolved in 1 ml of anhydrous acetonitrile, and then 4M HCl (0.49 ml, 1.95 mmol) was added dropwise. After the addition was completed, the reaction solution was stirred at 80 ° C for 3 hours. After the reaction was completed, the reaction solution was cooled to room temperature and quenched with saturated aqueous ammonium chloride solution. The reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine and then dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, and then gradient eluted by column chromatography to obtain compound 17-2.

Synthesis of compound S17

Compound 17-2 (123 mg, 0.50 mmol), compound 17-3 (124 mg, 0.50 mmol) and HATU (619 mg, 1.63 mmol) were dissolved in 3 ml of anhydrous N,N-dimethylformamide. Triethylamine (412 μl, 2.96 mmol) was slowly added to the stirring solution at room temperature, and stirred at room temperature after the addition was completed. After reacting for 6 hours, the reaction solution was extracted with ethyl acetate, and the organic phase was washed with saturated brine and then dried over anhydrous sodium sulfate. The organic phase was collected, concentrated, and then subjected to column chromatography gradient elution to obtain a white powder Like solid S17. ¹ H NMR(500MHz,chloroform-d)δ9.23(s,1H),8.06–7.93(m,2H),7.79(dd,J＝8.1,2.2Hz,1H),7.66–7.49(m,2H),7.42–7.22(m,2H),6.92(s,1H),4.02–3.19(m,8H ).

Preparation Example 18 Preparation of Compound S18:

Substrate 17-3 was replaced with N-(p-methoxyphenyl)-2,2-difluoro-4-piperidineacetamide, and the synthesis steps of compound S18 were the same as those of compound S17. ¹ H NMR(500MHz,chloroform-d)δ7.90–7.65(m,2H),7.66–7.51(m,2H),7.44–7.24(m,2H),6.92(t,J＝1.2Hz,2H),6.90(d,J＝1.6Hz,1H),5.88(s,1H),3.76(s,3H) ,3.72(dd,J＝10.8,8.1Hz,2H),3.60(dd,J＝10.9,8.2Hz,2H),2.47(s,J＝8.3Hz,1H),2.24(dt,J＝10.9,8.1Hz,2H),1.74(dt,J＝10.8,8.2Hz,2H).

Preparation Example 19 Preparation of Compound S19:

Substrate 17-3 was replaced with 1-methyl-5-indolecarboxylic acid, and the synthesis steps of compound S19 were the same as those of compound S17. ¹ H NMR (500 MHz, chloroform-d) δ7.87 (d, J=2.2 Hz, 1H), 7.59–7.53 (m, 2H), 7.53–7.48 (m, 2H), 7.45–7.40 (m, 1H), 7.40–7.34 (m, 2H), 6.92 (s, 1H), 6.30–6.26 (m, 1H), 3.85–3.75 (m, 8H), 3.69 (s, 3H).

Preparation Example 20 Preparation of Compound S20:

Substrate 1-1 was replaced with (E)-3-(4-fluorophenyl)acrylic acid, and substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S20 were the same as those of compound S1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.82–7.80 (m, 3H), 7.69 (s, 2H), 7.52 (d, J＝15.4 Hz, 1H), 7.37–7.23 (m, 5H), 3.78–3.56 (m, 8H).

Preparation Example 21 Preparation of Compound S21:

Substrate 1-1 was replaced with (E)-3-(4-trifluoromethylphenyl)acrylic acid, and substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S21 were the same as those of compound S1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.96 (d, J=8.0 Hz, 2H), 7.80–7.76 (m, 3H), 7.69 (s, 2H), 7.58 (d, J=15.4 Hz, 1H), 7.44 (d, J=15.5 Hz, 1H), 7.36 (d, J=8.2 Hz, 1H), 7.31 (dd, J=8.3, 1.7 Hz, 1H), 3.80 (br, 2H), 3.64–3.57 (m, 6H).

Preparation Example 22 Preparation of Compound S22:

Substrate 1-1 was replaced with (E)-3-(4-cyanophenyl)acrylic acid, and substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S22 were the same as those of compound S1. ¹ H NMR (400 MHz, chloroform-d) δ7.94(d,J＝8.1Hz,2H),7.88(d,J＝8.0Hz,2H),7.80(s,1H),7.69(s,2H),7.57(d,J＝15.3Hz,1H),7.46(d,J＝15.5Hz,1H),7.36(d,J＝8.2Hz,1H),7.32(s ,1H),3.79(s,2H),3.64(s,2H),3.56(s,4H).

Preparation Example 23 Preparation of Compound S23:

Substrate 1-1 was replaced with (E)-3-(4-chlorophenyl)acrylic acid, and substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S23 were the same as those of compound S1. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ7.81–7.73 (m, 3H), 7.69 (s, 2H), 7.56–7.42 (m, 3H), 7.37–7.26 (m, 3H), 3.78 (s, 2H), 3.66–3.60 (m, 2H), 3.56 (s, 4H).

Preparation Example 24 Preparation of Compound S24:

Substrate 1-1 was replaced with (E)-3-(4-dimethylaminophenyl)acrylic acid, and substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S24 were the same as those of compound S1. ¹ H NMR (500 MHz, chloroform-d) δ7.85 (d, J = 1.9 Hz, 1H), 7.56 (t, J = 1.0 Hz, 2H), 7.56–7.54 (m, 1H), 7.49–7.43 (m, 2H), 6.76 (d, J = 1.3 Hz, 1H), 6.74 (d, J = 1.3 Hz, 1H), 6.70 (d, J = 15.6 Hz, 1H), 6.60 (s, 2H), 3.76–3.71 (m, 4H), 3.69–3.65 (m, 4H), 2.98 (s, 6H).

Preparation Example 25 Preparation of Compound S25:

Substrate 1-1 was replaced with (E)-3-(4-trifluoromethoxyphenyl)acrylic acid, and substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S25 were the same as those of compound S1. ¹ H NMR (400 MHz, Methanol-d4) δ7.74 (d, J = 10.1 Hz, 3H), 7.59 (d, J = 15.4 Hz, 1H), 7.43 (d, J = 8.2 Hz, 1H), 7.37 (d, J = 8.5 Hz, 1H), 7.29 (d, J = 8.3 Hz, 2H), 7.18 (d, J = 15.8 Hz, 1H), 3.76 (s, 8H).

Preparation Example 26 Preparation of Compound S26:

Substrate 1-1 was replaced with (E)-3-(4-(3-fluoropropoxy)phenyl)acrylic acid, and substrate 1-4 was replaced with 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S26 were the same as those of compound S1. ^1. 60(s,2H),4.55(dt,J＝46.3,5.0Hz,2H),4.06(t,J＝6.8Hz,2H),3.82–3.66(m,8H),2.22–1.93(m,2H).

Preparation Example 27 Preparation of Compound S27:

Substrate 15-1 was replaced with 2-(1-(tert-butyloxycarbonyl)piperidin-4-yl)-2,2-difluoroacetic acid, and substrate 15-2 was replaced with benzo[d]thiazole-2,6-diamine. The synthesis steps of compound S27 were the same as those of compound 15. ¹ H NMR (500 MHz, chloroform-d) δ9.08 (s, 1H), 8.34 (dd, J = 1.8, 0.7 Hz, 1H), 7.66 (dd, J = 7.9, 0.7 Hz, 1H), 7.64 (dd, J = 7.7, 1.8 Hz, 1H), 7.56 (dd, J = 8.6, 0.6 Hz, 2H), 7.53 (d, J = 8.5 Hz, 2H), 7.4 5(dd,J＝15.6,0.7Hz,1H),6.57(s,2H),6.53(d,J＝15.6Hz,1H),3.73–3.59(m,4H),2.84(tt,J＝20.9,7.2Hz,1H),2.02(ddt,J＝12.4,9.8,7.4Hz,2H),1.79 (ddt,J＝12.4,9.7,7.5Hz,2H).

Preparation Example 28 Preparation of Compound S28:

Substrate 15-1 was replaced with 2-(1-(tert-butyloxycarbonyl)-3,3-difluoropiperidin-4-yl)acetic acid, and substrate 15-2 was replaced with benzo[d]thiazole-2,6-diamine. The synthesis steps of compound S28 were the same as those of compound 15. ¹ H NMR (500MHz, chloroform-d) δ9.29 (s, 1H), 8.26 (d, J＝2.0 Hz, 1H), 7.66 (d, J＝7.8 Hz, 1H), 7.60 (dd, J＝7.8, 1.9 Hz, 1H), 7.56 (dd, J＝8.6, 0.6 Hz, 2H), 7.53 (d, J＝8.5 Hz, 2H), 7.48–7.4 3(m,1H),6.66(d,J＝15.6Hz,1H),6.57(s,2H),3.97–3.83(m,2H),3.82–3.74(m,1H),3.65(ddd,J＝12.5,10.1,7.4Hz,1H),2.73–2.57(m,3H),2.07–1. 93(m,1H),1.90–1.72(m,1H).

Preparation Example 29 Preparation of Compound S29:

The substrate 1-tert-butyloxycarbonyl-piperazine was replaced by tert-butyl 1,4-diazepane-1-carboxylate, and the substrate 1-4 was replaced by 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S29 were the same as those of compound S1. 1H NMR (500MHz, chloroform-d) δ7.85 (d, J＝1.9Hz, 1H), 7.68 (d, J＝8.1Hz, 1H), 7.56 (dd, J＝8.6, 0.6H z,2H),7.53(d,J＝8.5Hz,2H),7.48–7.44(m,1H),7.44–7.42(m,1H),6.65(d,J＝15.6Hz,1H),6.60(s,2H),3.76–3.58(m,5H),3.58–3.46(m,2H),3.40 (dd,J＝7.5,6.8Hz,1H),1.94–1.80(m,2H).

Preparation Example 30 Preparation of Compound S30:

The substrate 1-tert-butyloxycarbonyl-piperazine was replaced by tert-butyl 2-methyl-1,4-diazepane-1-carboxylate, and the substrate 1-4 was replaced by 2-aminobenzothiazole-6-carboxylic acid. The synthesis steps of compound S30 were the same as those of compound S1. ¹ H NMR(500MHz,chloroform-d)δ7.91(d,J＝2.0Hz,1H),7.68(d,J＝8.0Hz,1H),7.56(dd,J＝8.6,0.7Hz,2H),7.53(d,J＝8.5Hz,2H),7.51(dd,J＝8.0,1.9Hz,1H),7.45(dt ,J＝15.6,0.6Hz,1H),6.73(d,J＝15.6Hz,1H),6.60(s,2H),4.16(qt,J＝7.7,5.8Hz,1H),3.72–3.64(m,1H),3.61(dd,J＝12.5,5.9Hz,1H),3.54– 3.43(m,2H),3.35(dt,J＝12.3,7.1Hz,1H),3.27–3.16(m,1H),1.90–1.70(m,2H),1.24(d,J＝7.7Hz,3H).

Preparation Example 31 Preparation of Compound S31:

Substrate 1-4 was replaced with 2-(methylamino)benzo[d]thiazole-6-carboxylic acid, and the synthesis steps of compound S31 were the same as those of compound S1. ¹ H NMR (500 MHz, chloroform-d) δ7.88 (d, J = 1.8 Hz, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.56 (dd, J = 8.6, 0.7 Hz, 2H), 7.53 (d, J = 8.5 Hz, 2H), 7.49–7.40 (m, 2H), 7.07 (q, J = 4.3 Hz, 1H), 6.70 (d, J = 15.6 Hz, 1H), 3.77–3.70 (m, 4H), 3.70–3.64 (m, 4H), 3.00 (d, J = 4.2 Hz, 3H).

Preparation Example 32 Preparation of Compound S32:

Substrate 1-4 was replaced with 2-(ethylamino)benzo[d]thiazole-6-carboxylic acid, and the synthesis steps of compound S32 were the same as those of compound S1. ¹ H NMR(500MHz,chloroform-d)δ7.88(d,J＝1.8Hz,1H),7.70(d,J＝7.9Hz,1H),7.56(dd,J＝8.6,0.6Hz,2H),7.53(d,J＝8.5Hz,2H),7.49–7.40(m,2H),7.19(t,J＝3.3 Hz,1H),6.70(d,J＝15.6Hz,1H),3.80–3.70(m,4H),3.70–3.63(m,4H),3.57(qd,J＝6.0,3.4Hz,2H),1.30(t,J＝6.0Hz,3H).

Preparation Example 33 Preparation of Compound S33:

Substrate 1-4 was replaced with 2-(isopropylamino)benzo[d]thiazole-6-carboxylic acid. The synthesis steps of compound S33 were the same as those of compound S1. 1H NMR (500 MHz, chloroform-d) δ7.88 (d, J = 1.9 Hz, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.56 (dd, J = 8.6, 0.6 Hz, 2H), 7.53 ( d,J＝8.5Hz,2H),7.48–7.42(m,2H),6.70(d,J＝15.6Hz,1H),6.16(d,J＝6.2Hz,1H),4.04(dp,J＝6.4,5.4Hz,1H),3.80–3.70(m,4H),3.70–3.62(m,4H), 1.25(d,J=5.3Hz,6H).

Preparation Example 34 Preparation of Compound S34:

Substrate 1-4 was replaced with 2-(tert-butylamino)benzo[d]thiazole-6-carboxylic acid, and the synthesis steps of compound S34 were the same as those of compound S1. ¹ H NMR (500 MHz, chloroform-d) δ7.88 (d, J = 1.9 Hz, 1H), 7.70 (d, J = 7.9 Hz, 1H), 7.56 (dd, J = 8.6, 0.6 Hz, 2H), 7.53 (d, J = 8.5 Hz, 2H), 7.48–7.41 (m, 2H), 6.70 (d, J = 15.6 Hz, 1H), 5.85 (s, 1H), 3.76–3.72 (m, 4H), 3.69–3.63 (m, 4H).

Experimental Example 1. Evaluation of the GPR183 Inhibitory Activity of Compounds

1. Experimental purpose:

Using the Glosensor cAMP assay, we screened out compounds that can effectively inhibit GPR183 activity 2. Test method:

One day in advance, HEK293 cells to be transfected the next day were inoculated in a 6 cm dish. The inoculation principle was to ensure that the cell density on the next day was 80-90%. The next day, the transfection plasmids GPR183 and pGloSensor ^TM -22F cAMP were transfected at a ratio of 1:1 using serum-free DMEM medium. The ratio of the volume of the transfection reagent PEI to the total amount of transfected plasmids was 3:1, and the total amount of plasmid transfection in the 6 cm dish was 2ug. After 5-6 hours, the transfection solution was discarded, the cells were washed once with 1x PBS, and serum-containing DMEM medium was added for overnight culture to allow the plasmid to be fully expressed. After overnight culture, the culture medium was discarded, the cells were washed once with 1x PBS, 500ul of trypsin was added for digestion for 30s, the trypsin was aspirated, and 3ml of _CO2- independent culture medium was added to resuspend the cells. Then, GloSensor substrate (Promega) was added under light-proof conditions and mixed evenly to allow the substrate to fully contact the cells. After the cell suspension is evenly mixed, the cells are evenly spread into a 384-well plate with 30ul of cell suspension in each well. If the agonist is detected, for Gi-coupled receptors, forskolin is mixed with agonist 7α,25-OHC at different concentration gradients in a 1:1 ratio, and then 30ul of the mixed solution is added to the 384-well plate that has already read the background; if the antagonist is detected, for Gi-coupled receptors, 15ul of antagonists at different concentration gradients are first added to the cell suspension and incubated for 20 minutes, and then a mixture of forskolin and agonist EC ₈₀ is added. Use a LUMIstar Omega chemiluminescent microplate reader to quantify the luminescence intensity;

3. The experimental results are shown in Table 1.

Table 1 Cell activity test results

Selected compounds of the present invention were screened in the above test method, and the results are summarized in the above table as Groups A, B and C. In this document, Group "A" refers to _IC50 values between >0.1 nM and ≤2 nM, Group "B" refers to _IC50 values between >2 nM and ≤10 nM, and Group "C" refers to _IC50 values between >10 nM and ≤100 nM.

Experimental Example 2. In vivo pharmacokinetic study

1. Experimental purpose:

This experiment aims to investigate the pharmacokinetic characteristics of the compounds of the present invention in mice after oral administration and intravenous injection.

2. Test method:

Six mice were divided into two groups and given the test compound by gavage and tail vein injection, respectively. Blood samples of about 0.05 ml were collected from the cheeks of the intravenous injection group at 5min, 15min, 30min, 1h, 2h, 4h, 6h, 8h, and 24h after administration; blood samples of about 0.05 ml were collected from the gavage group at 5min, 15min, 30min, 1h, 2h, 4h, 6h, 8h, and 24h after administration. The concentration of the test compound in mouse plasma samples was determined by LC-MS/MS, and the pharmacokinetic parameters were calculated using WinNolin software.

3. The experimental results are shown in Table 2.

Table 2 Comparison of PK properties of compounds and NIBR189 in mice

4. Experimental conclusion

The compounds of the present invention have good pharmacokinetic characteristics in mice. Compared with the positive compound NIBR189, under intravenous administration, the plasma exposure (AUC _0-∞ ) of the representative compound S10 is much higher than that of NIBR189 (1918 vs 966hr*ng/ml), and the clearance rate is much lower than that of NIBR189 (523 vs 1038ml/hr/kg). Similarly, under oral administration, the plasma exposure (AUC _{0- ∞} ) of the representative compound S10 is much higher than that of NIBR189 (4092 vs 2070hr*ng/ml), and the half-life is also slightly longer than that of NIBR189. Therefore, the pharmacokinetic characteristics of the representative compound S10 are better than those of NIBR189.

Experimental Example 3. Evaluation of the inhibitory activity of representative compound S10 on GPR183

1. Experimental purpose:

The inhibitory activity of representative compound S10 on GPR183 was tested by Glosensor cAMP and CRE reporter gene assays.

2. Test method:

(1) Glosensor cAMP detection method:

Same as above.

(2) CRE reporter gene detection method:

One day in advance, use a 6 cm dish to inoculate the HEK293 cells to be transfected the next day. The inoculation principle is to ensure that the cell density on the second day is 80-90%; the next day, use serum-free DMEM medium to transfect the plasmid GPR183 and CRE reporter gene plasmid at a ratio of 1:1, and the ratio of the volume of transfection reagent PEI to the total amount of transfected plasmid is 3:1, where the total amount of plasmid transfection in the 6 cm dish is 2ug; 5-6h later, pour out the transfection solution, wash once with 1x PBS, digest with trypsin, and resuspend in complete medium to a cell suspension with a cell density of 300,000/ml, inoculate in a 96-well plate, 100ul/well, and wait for the cells to adhere to the wall. Incubate with a certain concentration of antagonist for 20 minutes, then add a certain concentration of agonist 7α, 25-OHC and forskolin mixed solution and incubate for 4 hours. After the incubation, add 100ul firefly luciferase reporter gene cell lysate to each well and place it at -80℃ overnight. The next day, take out the plate with lysate from -80℃, thaw it on a shaker at 120r for 40 minutes, then place it in a high-speed refrigerated centrifuge at 3700rpm and centrifuge it at room temperature for 10 minutes. After the centrifugation, aspirate 35ul of supernatant and spread it in a 384-well plate, add 35ul of luciferin substrate, and immediately put it into the LUMIstar Omega chemiluminescence microplate reader to read the value.

3. The experimental results are shown in Figure 1.

4. Experimental conclusion

In both the Glosensor cAMP test system and the CRE reporter gene test system, the representative compound S10 showed more potent GPR183 inhibitory activity compared with NIBR189.

Experimental Example 4. Test of the hERG potassium channel effect of representative compound S10

1. Experimental purpose:

This experiment aims to investigate the possible hERG potassium channel toxicity of the representative compound S10.

2. Test method:

The hERG potassium channel effect of the compound was tested using the fully automated patch clamp Qpatch detection technology. CHO-hERG cells were cultured to a cell density of 60-80%, the culture medium was removed, washed with 7 mL PBS, and then treated with trypsin. After complete digestion, the culture medium was added to neutralize, then centrifuged and the supernatant was aspirated, and then the culture medium was added to resuspend. The compound was then diluted with the Bravo instrument to obtain six different concentrations. DMSO solution. (DMSO content does not exceed 0.2%, at this concentration DMSO has no effect on hERG potassium channels), and then prepare solutions (including intracellular fluid and extracellular fluid).

The single-cell high-impedance sealing and whole-cell pattern formation process are all automatically completed by the Qpatch instrument. After obtaining the whole-cell recording mode, the cell is clamped at -80 mV. Before giving a 5-second +40 mV depolarizing stimulus, a 50-millisecond -50 mV pre-voltage is given, and then repolarized to -50 msec and maintained for 5 seconds, and then returned to -80 mV. This voltage stimulus is applied every 15 seconds. After recording for two minutes, the extracellular solution is given for five minutes of recording, and then the drug administration process begins. The compound concentration starts from the lowest test concentration, and each test concentration is given for 2.5 minutes. After all concentrations are given continuously, 3 μL Cisapride is given based on the positive control compound. At least two cells (n≥2) are tested for each concentration.

3. Experimental results are shown in Figure 2. 4. Experimental conclusion

In the hERG potassium channel action test, the hERG IC ₅₀ value of compound S10 is 8.71 μM, which is better than NIBR189 (hERG IC ₅₀ = 1.6 μM) reported in the previous document (WO2023066190). Therefore, compared with NIBR189, compound S10 has lower hERG toxicity.

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims (10)

A compound, characterized in that the compound is a compound represented by formula I, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof,
Wherein, ring A is a saturated or unsaturated 4-10 membered heterocycloalkyl containing 1, 2 or 3 N; n is selected from the following group: 0, 1, 2, 3; Each R ₁ , R ₂ , and R ₃ is independently selected from the following group: hydrogen, deuterium, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C4 alkyl-(C＝O)-NH-, -N(C1-C6 alkyl) ₂ , -NH(C1-C6 alkyl), -NH ₂ , wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: halogen, hydroxyl, amino; Y is a straight bond or a C1-C5 alkylene group; When Y is a straight bond, T is R ₄ and R ₅ are each independently selected from the following group: H, C1-C6 alkoxy; or, R ₄ and R ₅ together with the phenyl carbon to which they are connected form a substituted or unsubstituted 3-6 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 3-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: hydrogen, C1-C4 alkyl, halo C1-C4 alkyl, halogen, =O, hydroxyl, thiol, amino, -NH(C1-C6 alkyl), -N(C1-C6 alkyl) ₂ ; When Y is C1-C5 alkylene, T is R ₄ and R ₅ are each independently selected from the following group: H, C1-C6 alkoxy; or, R ₄ and R ₅ together with the phenyl carbon to which they are connected form a substituted or unsubstituted 3-6 membered heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 3-6 membered heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O or S, wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, halogen, =O, hydroxyl, mercapto, amino, -NH(C1-C6 alkyl), -N(C1-C6 alkyl) ₂ ; R' is selected from the following group: hydrogen, deuterium, halogen, hydroxyl, cyano, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C4 alkyl-(C=O)-NH-, wherein the substitution independently refers to substitution by 1, 2 or 3 substituents selected from the following group: halogen, hydroxyl, amino; m is selected from the following group: 1, 2, 3; The additional condition is that the compound is not The compound according to claim 1, characterized in that Ring A is a saturated 4-8-membered heterocycloalkyl group containing 2 N atoms, and Ring A is a monocyclic ring; n is 0; Each R ₁ , R ₂ , and R ₃ is independently selected from the group consisting of hydrogen, halogen; Y is a straight bond, T is for X ₁ , X ₂ , and X ₃ are each independently selected from the group consisting of N, CR ₆ , C(R ₆ ) ₂ , NR ₆ , S, and O; Each R ₆ is independently selected from the group consisting of hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, halogen, =0, hydroxyl, mercapto, and amino. The compound according to claim 2, characterized in that Select from the following group:
The compound according to claim 1, characterized in that Ring A is a saturated 6-10 membered heterocycloalkyl group containing 2 N atoms, and Ring A is a bridged ring; n is 0; Each R ₁ , R ₂ , and R ₃ is independently selected from the group consisting of hydrogen, halogen; Y is a straight bond, T is _R4 and _R5 are each independently selected from the group consisting of H, C1-C6 alkoxy. The compound according to claim 1, characterized in that Ring A is a saturated 4-8-membered heterocycloalkyl group containing 1 N, and Ring A is a monocyclic ring; n is selected from the following group: 0, 1, 2, 3; Each R ₁ , R ₂ , and R ₃ is independently selected from the group consisting of hydrogen, halogen; Y is C1-C5 alkylene, T is _R4 and _R5 are each independently selected from the group consisting of H, C1-C6 alkoxy; R' is selected from the group consisting of hydrogen, halogen; m is selected from the following group: 1, 2, 3. The compound according to claim 1, characterized in that Ring A is a saturated 4-8-membered heterocycloalkyl group containing 2 N atoms, and Ring A is a monocyclic ring; n is 0; Each of R ₁ , R ₂ , and R ₃ is independently halogen; Y is a straight bond, T is for X ₁ , X ₂ , and X ₃ are each independently selected from the group consisting of N, CR ₆ , C(R ₆ ) ₂ , NR ₆ , S, and O; Each R ₆ is independently selected from the group consisting of hydrogen, C1-C4 alkyl, halogenated C1-C4 alkyl, halogen, =0, hydroxyl, mercapto, and amino. The compound according to claim 1, characterized in that the compound is selected from the group consisting of:


A pharmaceutical composition, characterized in that it comprises a pharmaceutically acceptable carrier and one or more safe and effective amounts of the compound according to claim 1. A use of the compound according to claim 1, characterized in that it is used to prepare a drug for preventing and/or treating GPR183-related diseases. The use according to claim 9, characterized in that the GPR183-related disease is selected from the group consisting of tumors, inflammation, autoimmune diseases, metabolic diseases, pain, bacterial infections, and viral infections.