CN107827837B

CN107827837B - Sphingosine-1-phosphate receptor modulator compounds, and preparation method and application thereof

Info

Publication number: CN107827837B
Application number: CN201711168722.8A
Authority: CN
Inventors: 张健; 马仕珉; 王刚; 王飞; 欧洋
Original assignee: SUZHOU LANXITE BIOTECHNOLOGY CO Ltd
Current assignee: SUZHOU LANXITE BIOTECHNOLOGY CO Ltd
Priority date: 2017-11-21
Filing date: 2017-11-21
Publication date: 2021-09-24
Anticipated expiration: 2037-11-21
Also published as: CN107827837A; WO2019101045A1

Abstract

The invention relates to sphingosine-1-phosphate receptor modulator compounds, a preparation method and application thereof. Specifically provided are compounds represented by the following formula (I), racemates, stereoisomers, tautomers, solvates, hydrates or pharmaceutically acceptable salts thereof:

Description

Sphingosine-1-phosphate receptor modulator compounds, and preparation method and application thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to a sphingosine-1-phosphate receptor modulator compound, and a preparation method and application thereof.

Background

Sphingosine-1-phosphate (S1P) is a sphingomyelin metabolite and is involved in cell growth, proliferation, migration, and other processes. The S1P receptor belongs to G protein coupled receptor, and comprises S1P₁、S1P₂、S1P₃、S1P₄、S1P₅Five subtypes. Wherein, S1P₁The receptor agonist can reduce the number of peripheral blood lymphocytes and play a role in immunoregulation. The first marketed S1P receptor agonist drug Fingolimod (FTY 720) has proven effective in pathological models of various autoimmune diseases, and is clinically approved for the treatment of multiple sclerosis. However, the drug has agonistic effects on multiple subtypes of the S1P receptor, which is on S1P₃Receptor agonism causes bradycardia side effects, resulting in limited clinical use. Therefore, it is necessary to develop a composition having S1P₁A receptor selective immunomodulator.

Disclosure of Invention

In order to solve the above problems, the present invention provides a compound represented by formula (I), a racemate, a stereoisomer, a tautomer, a solvate, a hydrate, or a pharmaceutically acceptable salt thereof:

wherein R is₁Selected from H, unsubstituted or substituted by one or more R_aSubstituted of the following groups: c_1-8Alkyl radical, C_3-8Cycloalkyl, heterocyclyl, aryl or heteroaryl;

R₂is-CN;

R₃is selected from-NR'₂Or aza C_3-8A cycloalkyl group; wherein, said aza C_3-8Cycloalkyl radicals substituted by one or more-R₆COOR₇Substituted; r' may be the same or different, and is optionally: H. c_1-8Alkyl, -R₆COOR₇Wherein at least one of R' is-R₆COOR₇(ii) a Wherein R is₆Selected from chemical bonds or C_1-8Alkylene radical, R₇Is selected from H or C_1-8An alkyl group;

R_aselected from-F, -Cl, -Br, -I, -CN, -OH, ═ O or the following groups substituted by-F, -Cl, -Br, -I, -CN, -OH, ═ O: c_1-8Alkyl radical, C_3-8Cycloalkyl, heterocyclyl, aryl or heteroaryl.

According to an embodiment of the compounds of the invention, wherein R₁Selected from unsubstituted or substituted by one or more R_aSubstituted of the following groups: c_1-6Alkyl radical, C_3-6A cycloalkyl group; the R is_aSelected from-F, -Cl, -Br, -I, -CN or the following group substituted by-F, -Cl, -Br, -I, -CN: c_1-6Alkyl radical, C_3-6A cycloalkyl group;

as an example, R₁Selected from unsubstituted or substituted by one or more R_aSubstituted of the following groups: methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, cyclopentyl, n-hexyl, isohexyl, cyclohexyl; the R is_aSelected from-F, -Cl, -Br, -I or-CN;

as an example, R₃Is selected from

Wherein R is₄、R₅Is selected from H or C_1-6An alkyl group; l is selected from C_1-6Alkylene, m and n are identical or different and are each independently selected from the group consisting of integers from 1 to 3, R₆Selected from chemical bonds or C_1-8An alkylene group;

as an example, R₃Selected from-azetidinyl-R₆COOR₇-azacyclopentyl-R₆COOR₇or-azacyclohexyl-R₆COOR₇。

According to an embodiment of the compounds of the present invention, the pharmaceutically acceptable salts of the compounds are selected from salts of inorganic or organic acids with salifiable sites in the compounds (e.g., salifiable N-up);

the inorganic acids include, but are not limited to: hydrochloric acid, sulfuric acid, hydrobromic acid, phosphoric acid, hydrofluoric acid, hydroiodic acid, pyrosulfuric acid, or nitric acid;

such organic acids include, but are not limited to: citric acid, maleic acid, succinic acid, acetic acid, malic acid, tartaric acid, methanesulfonic acid, benzenesulfonic acid, formic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2- (4-hydroxybenzoyl) benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, p-toluenesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, stearic acid, lactic acid, oxalic acid, malonic acid, adipic acid, alginic acid, fumaric acid, D-gluconic acid, succinic acid, pyruvic acid, trifluoroacetic acid, propionic acid, benzoic acid, cinnamic acid, and mixtures thereof, Mandelic acid, ascorbic acid, glucoheptylic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

Preferably, the pharmaceutically acceptable salt of the compound is selected from the hydrochloride salt.

As an embodiment of the present invention, the compound of formula (I) is selected from the group comprising, but not limited to, the following compounds or salts thereof:

the invention also provides a preparation method of the compound shown in the formula (I), which comprises the following steps: reacting the compound of formula (I-4) with the compound of formula (I-5) to obtain the compound of formula (I)

Wherein R is₁、R₂、R₃Having the definitions as described above.

Preferably, when R is₃Group R in (1)₇In the case of alkyl, the resulting compound of the formula (I) may be subjected to waterDecomposing and reacting to obtain R₇A compound of formula (I) which is H.

Preferably, the compound of formula (I-4) can be prepared by a process comprising:

wherein R is₁、R₂、R₃Having the definitions set out above;

1) reacting 4-cyanobenzaldehyde with ethylene glycol to obtain a compound of formula (I-1);

2) reacting the compound of the formula (I-1) obtained in the step 1) with hydroxylamine hydrochloride to obtain a compound of a formula (I-2);

3) reacting the compound of the formula (I-2) obtained in the step 2) with a compound of a formula (II) to obtain a compound of a formula (I-3), and then reacting under an acidic condition to obtain a compound of a formula (I-4).

Optionally, the preparation method further comprises reacting the compound of formula (I) with a salt-forming reagent to form a pharmaceutically acceptable salt thereof.

Preferably, the salifying agent may be an organic or inorganic acid;

the organic or inorganic acid has the definition as described above, for example the inorganic acid is selected from hydrochloric acid or a solution thereof.

If protection is desired, any functional group in the compounds of formula (II), formula (I-3), formula (I-4), formula (I-5) may be protected, and then, if necessary, the protecting group may be removed.

The term "compound of the invention" as used in the present invention includes one or more compounds of formula (I), racemates, stereoisomers, tautomers, solvates, hydrates or pharmaceutically acceptable salts thereof.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention.

Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

Such pharmaceutically acceptable carriers include, but are not limited to: one or more of fillers, diluents, disintegrants, lubricants, glidants, binders, solubilizers, surfactants, emulsifiers, preservatives, antioxidants, flavoring agents, colorants, osmotic pressure regulators, pH regulators, proppants.

Preferably, in the pharmaceutical composition, the mass of the compound of the present invention accounts for 0.01-99.99% of the total mass of the pharmaceutical composition.

Preferably, the pharmaceutical composition is used alone or in combination with other therapies or pharmaceutical preparations, and the dosage and frequency of administration are determined in combination with factors such as the route of administration, the type of condition, the age of the patient, and whether or not to take other drugs. In general, the dosage range is from 0.001mg to 100mg per kg of body weight per day, with the required dosage being provided in single or divided doses per day.

The pharmaceutical composition can be administered by oral administration, injection, infusion, drip, inhalation, or application.

The invention also provides a preparation containing the pharmaceutical composition.

Dosage forms of the formulation include, but are not limited to: liquid dosage forms, solid dosage forms, or semi-solid dosage forms.

The liquid dosage form can be oral solution preparation, injection, infusion solution, lotion, drop, liniment, aerosol, etc.; the solid dosage form can be tablet, capsule, granule, powder, pill, suppository, patch, etc.; the semisolid dosage form may be an ointment, gel, paste, and the like.

The dosage forms also include common preparations, sustained release preparations, controlled release preparations, targeted preparations and various microparticle drug delivery systems.

The invention also provides a treatment S1P₁A method of treating a receptor-related disorder comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition thereof.

The S1P₁Receptor-related disorders include, but are not limited to: autoimmune diseases, lymphocyte-mediated diseases, inflammatory diseases, bacterial infections, fungal infections, viral infections or cancer, etc.

By way of example, the method of treatment is the administration of a therapeutically effective amount of a compound of the invention or a pharmaceutical composition thereof as described above to an individual suffering from multiple sclerosis, and/or inflammatory bowel disease, and/or crohn's disease, and/or ulcerative colitis, and/or systemic lupus erythematosus, and/or rheumatoid arthritis, and/or psoriasis, and/or transplant rejection.

The subject may be a human or other mammal.

The invention also provides the use of a compound of the invention or a pharmaceutical composition thereof in the preparation of a medicament for the treatment of S1P₁Use in medicine for a receptor-related disorder.

Preferably, the use can be in the preparation of a medicament for the treatment of inflammatory bowel disease, and/or crohn's disease, and/or ulcerative colitis, and/or systemic lupus erythematosus, and/or rheumatoid arthritis, and/or psoriasis, and/or multiple sclerosis, and/or transplant rejection.

Has the advantages that:

the invention provides a S1P with a brand-new structure₁Receptor modulators, compounds of this type with the already reported S1P₁Receptor modulator Compounds for S1P₁The agonist strength and selectivity of the receptor are improved, and the agonist is S1P₃Have little agonistic activity and have higher immunosuppressive activity and metabolic stability, for example the percentage of lymphocyte reduction may exceed 60%. In a biological activity experiment, the compound is proved to have obvious treatment effect on the ulcerative colitis and appropriate in vivo bioavailability and can become S1P₁A therapeutic agent for a receptor-related disease.

Definition and description of terms:

unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter of the application. In this application, the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the term "comprising" as well as other forms, such as "includes," "including," and "containing," are not limiting.

The term "alkyl" refers to a straight or branched alkyl group having 1 to 8, preferably 1 to 6, carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sec-butyl, pentyl, neopentyl.

The term "aryl" is understood to mean preferably a mono-, bi-or tricyclic hydrocarbon ring having a monovalent aromatic or partially aromatic character of 6 to 20 carbon atoms, preferably "C_6-14Aryl ". The term "C_6-14Aryl "is to be understood as preferably meaning a mono-, bi-or tricyclic hydrocarbon ring having a monovalent or partially aromatic character with 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms (" C_6-14Aryl group "), in particular a ring having 6 carbon atoms (" C₆Aryl "), such as phenyl; or biphenyl, or is a ring having 9 carbon atoms ("C₉Aryl), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C₁₀Aryl radicals), such as tetralinyl, dihydronaphthyl or naphthyl, or rings having 13 carbon atoms ("C₁₃Aryl radicals), such as the fluorenyl radical, or a ring having 14 carbon atoms ("C)₁₄Aryl), such as anthracenyl.

The term "heteroaryl" is understood to mean monocyclic, bicyclic and tricyclic ring systems containing 5 to 20 ring atoms, 5 to 14 ring atoms, or 5 to 12 ring atoms, or 5 to 10 ring atoms, or 5 to 6 ring atoms, at least one of which is aromatic, and at least one of which contains one or more heteroatoms (e.g., N, O, S, Se, etc.), wherein each ring system contains a ring of 5 to 7 atoms with one or more attachment points to the rest of the molecule. The heteroaryl group is optionally substituted with one or more substituents described herein. In some embodiments, a heteroaryl group of 5-10 atoms contains 1,2,3, or 4 heteroatoms independently selected from O, S, Se and N. In other embodiments, a 5-6 atom heteroaryl group contains 1,2,3, or 4 heteroatoms independently selected from O, S, Se and N.

Examples of monocyclic rings of heteroaryl groups include, but are not limited to, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl and the like and their benzo derivatives, such as benzofuranyl, benzothienyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl, isoquinolyl, and the like; or azocinyl, indolizinyl, purinyl and the like and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like.

The term "heterocyclyl" means a monocyclic, bicyclic or tricyclic ring system in which one or more atoms in the ring are independently optionally substituted with a heteroatom, the ring may be fully saturated or contain one or more unsaturations, but is not aromatic, having one or more points of attachment to other molecules. One or more ring hydrogen atoms may be independently unsubstituted or substituted with one or more substituents described herein. Some of these embodiments are "heterocyclyl" groups which are monocyclic of 3 to 7 atoms, or bicyclic of 7 to 10 atoms, containing 1 to 5, preferably 1 to 3 heteroatoms selected from N, O, S and Se. In particular, the heterocyclic group may include, but is not limited to: 4-membered rings such as azetidinyl, oxetanyl; 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or a 6-membered ring such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl; or a 7-membered ring such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclyl group may be bicyclic, for example but not limited to a 5,5 membered ring, such as a hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a 5,6 membered bicyclic ring, such as a hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring. The nitrogen atom containing ring may be partially unsaturated, i.e. it may contain one or more double bonds, such as but not limited to 2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 4, 5-dihydrooxazolyl or 4H- [1,4] thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolyl, 1, 3-benzoxazolyl, 1, 3-benzodioxolyl.

Unless otherwise indicated, heterocyclyl, heteroaryl include all possible isomeric forms thereof, e.g. positional isomers thereof. Thus, for some illustrative, non-limiting examples, pyridyl or pyridinylene includes pyridin-2-yl, pyridinylene-2-yl, pyridin-3-yl, pyridinylene-3-yl, pyridin-4-yl, and pyridinylene-4-yl; thienyl or thienylene includes thien-2-yl, thien-3-yl and thien-3-yl.

As used herein, reference to the term "subject", "patient" or "individual" refers to an individual suffering from a disease, disorder or condition, and the like, including mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs, and the like. Examples of non-human mammals include, but are not limited to, birds, fish, and the like. In one embodiment related to the methods and compositions provided herein, the mammal is a human.

As used herein, the term "treating" and other similar synonyms include alleviating, or ameliorating a symptom of a disease or disorder, preventing other symptoms, ameliorating, or preventing an underlying metabolic cause of a symptom, inhibiting a disease or disorder, e.g., arresting the development of a disease or disorder, alleviating a disease or disorder, ameliorating a disease or disorder, alleviating a symptom of a disease or disorder, or discontinuing a symptom of a disease or disorder, and further, the term encompasses prophylactic purposes. The term also includes obtaining a therapeutic effect and/or a prophylactic effect. The therapeutic effect refers to curing or ameliorating the underlying disease being treated. In addition, a cure or amelioration of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, e.g., an improvement in the condition of the patient is observed, although the patient may still be affected by the underlying disease. For prophylactic effect, the composition can be administered to a patient at risk of developing a particular disease, or to a patient presenting with one or more physiological symptoms of the disease, even if a diagnosis of the disease has not yet been made.

The terms "effective amount," "therapeutically effective amount," or "pharmaceutically effective amount" as used herein, refer to an amount of at least one agent or compound that is sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

The terms "administering," "administration," "administering," and the like, as used herein, refer to a method capable of delivering a compound or composition to a desired site for biological action. These methods include, but are not limited to, oral routes, via the duodenal route, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), topical and rectal administration. Administration techniques useful for the compounds and methods described herein are well known to those skilled in the art. In preferred embodiments, the compounds and compositions discussed herein are administered orally.

The term "acceptable" as used herein with respect to a formulation, composition or ingredient means that there is no long-term deleterious effect on the general health of the subject being treated.

The term "pharmaceutically acceptable" as used herein refers to a substance (e.g., carrier or diluent) that does not affect the biological activity or properties of the compounds of the present application, and is relatively non-toxic, i.e., the substance can be administered to an individual without causing an adverse biological response or interacting in an adverse manner with any of the components contained in the composition.

The term "pharmaceutical composition" as used herein refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical ingredient including, but not limited to, carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and/or excipients.

The term "carrier" as used herein refers to a relatively non-toxic chemical compound or agent that facilitates the introduction of the compound into a cell or tissue.

Detailed Description

The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

The positive control used in examples 2 to 6 was Ozanimod developed by Receptos corporation, made by oneself. Ozanimod is an alternative S1P developed by Receptos₁A receptor modulator. It is currently in the clinical study stage of multiple immune diseases, and data show that it has a high S1P₁Receptor selectivity and good pharmacokinetic properties.

The structure of Ozanimod is shown below:

preparation of the Compound of example 1 and Structure analysis data

(1) Preparation of intermediate I-1

Intermediate I-1: intermediate 4-formylbenzonitrile (6.0g,45.76mmol), ethylene glycol (9.32ml,183.02mmol) and p-toluenesulfonic acid (626mg, 3.66mmol) were dissolved in 240ml of toluene, heated under reflux for 2h and partitioned with a trap. Cooling to room temperature, evaporating off the solvent, dissolving the residue in 300ml of ethyl acetate, washing with water, drying, and carrying out column chromatography, wherein the eluent is petroleum ether: 20 parts of dichloromethane: 1, 5.8g of white solid is obtained, the yield is 71.2%.¹H NMR(400MHz,DMSO-d₆)δ7.87(d,J＝8.4Hz,2H),7.62(d,J＝8.4Hz,2H),5.83(s,1H),3.94-4.08(m,4H).MS(ESI)m/z(M+H)⁺176.8。

(2) Preparation of intermediate I-2

Intermediate I-2: intermediate I-1(5.60g,31.96mmol) was dissolved in 240ml of anhydrous methanol, and hydroxylamine hydrochloride (7.78g, 111.88mmol) and sodium hydrogencarbonate (10.74g, 127.86mmol) were added successively under stirring, followed by heating and refluxing for 10 hours. Cooling, suction filtering, washing the filter cake with anhydrous methanol, and combining the filtrates. Adding distilled water into the filtrate, extracting with ethyl acetate, washing with water, drying, and carrying out column chromatography, wherein an eluent is dichloromethane: methanol 20:1, obtaining 4.3g of white solid with the yield of 65.4 percent.¹H NMR(400MHz,DMSO-d₆)δ9.66(s,1H),7.67-7.69(m,2H),7.41-7.43(m,2H),5.82(s,2H),5.74(s,1H),3.93-4.07(m,4H).MS(ESI)m/z(M+H)⁺209.4。

(3) Preparation of intermediates I-3-1, I-3-2

Intermediate I-3-1 (R)₁Is isopropyl, R₂CN): 3-cyano-4-isopropoxybenzeneFormic acid (4.84g,23.58mmol), 1-hydroxybenzotriazole (2.9g,21.42mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (4.1g,21.42mmol), potassium carbonate (4.44g,32.14mmol) were dissolved in 240ml of DMF, stirred at room temperature, intermediate I-2(5.2g,21.42mmol) was added, and the reaction was heated. Cooling, filtering, washing filter cake with ethyl acetate, and merging filtrate. Adding distilled water into the filtrate, extracting with ethyl acetate, washing with water, drying, concentrating, and performing column chromatography, wherein an eluent is petroleum ether: ethyl acetate 4:1 gave 3.58g of a white solid in 53.1% yield.¹H NMR(400MHz,DMSO-d₆)δ8.46(s,1H),8.37(d,J＝8.9Hz,1H),8.10(s,2H),7.64(d,J＝8.2Hz,2H),7.52(d,J＝9.1Hz,1H),5.82(s,1H),4.96(p,J＝6.0Hz,1H),3.93-4.16(m,4H),1.38(d,J＝6.0Hz,6H)；MS(ESI)m/z 378.4。

Intermediate I-3-2 (R)₁Is cyclopentyl, R₂CN): the same preparation as described for intermediate I-3-1 was carried out using 3-cyano-4-cyclopentyloxybenzoic acid instead of 3-cyano-4-isopropoxybenzoic acid to give 6.7g of a white solid in 61.1% yield.¹H NMR(400MHz,DMSO-d₆)δ8.50(d,J＝2.4Hz,1H),8.41(dd,J＝9.0,2.2Hz,1H),8.10-8.12(m,2H),7.65-7.67(m,2H),7.52(d,J＝8.8Hz,1H),5.84(s,1H),5.14-5.18(m,1H),3.96-4.12(m,4H),1.99-2.06(m,2H),1.63-1.82(m,6H).MS(ESI)m/z(M+H)⁺404.5。

(4) Preparation of intermediates I-4-1, I-4-2

Intermediate I-4-1 (R)₁Is isopropyl, R₂CN): intermediate I-3-1(4.60g,12.18mmol) was dissolved in 60ml of acetone, and 2N hydrochloric acid solution was added with stirring and heated at 45 ℃ for 3 hours. The reaction was cooled to room temperature, filtered under reduced pressure and dried to give 2.28g of a white solid with a yield of 56.3%.¹H NMR(400MHz,DMSO-d₆)δ10.11(s,1H),8.48(d,J＝2.4Hz,1H),8.39(dd,J＝9.0,2.2Hz,1H),8.28(d,J＝8.4Hz,2H),8.10(d,J＝8.0Hz,2H),7.54(d,J＝9.2Hz,1H),4.97(hept,J＝6.0Hz,1H),1.38(d,J＝6.0Hz,6H).MS(ESI)m/z(M+H)⁺334.1。

Intermediate I-4-2 (R)₁Is cyclopentyl, R₂CN): the intermediate I-3-1 was replaced with the intermediate 1-3-2 in the same manner as described for the preparation of the intermediate I-4-1 to give 4.45g of a white solid with a yield of 48.5%.¹1H NMR(400MHz,DMSO-d₆)δ10.12(s,1H),8.52(d,J＝2.4Hz,1H),8.42(dd,J＝8.8,2.4Hz,1H),8.29-8.32(m,2H),8.10--8.14(m,2H),7.53(d,J＝9.2Hz,1H),5.15-5.19(m,1H),1.98-2.07(m,2H),1.63-1.83(m,6H).MS(ESI)m/z(M+H)⁺360.8。

(5) Preparation of target compound LK-S1P-001

(4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazol-3-yl) benzyl) glycinate hydrochloride

Intermediate I-4-1(400mg,1.2mmol) was dissolved in 70ml dichloromethane, glycine methyl ester hydrochloride (222mg,1.8mmol), glacial acetic acid (0.28ml,4.80mmol) and N, N-diisopropylethylamine (0.3ml,1.8mmol) and 15ml methanol were added, reacted at room temperature for 3h, then added with sodium cyanoborohydride (76mg, 1.2mmol) and reacted under argon for 10 h. Saturated sodium bicarbonate solution was added, extracted 3 times with dichloromethane (3 × 50mL), washed with water, dried, separated by column chromatography eluting with dichloromethane: methanol 20:1 gave 298mg of a white solid in 61.1% yield.¹H NMR(400MHz,DMSO-d₆)δ8.49(m,1H),8.39(dt,J＝9.2,1.7Hz,1H),8.03(d,J＝8.0Hz,2H),7.53-7.56(m,3H),4.97(hept,J＝6.0Hz,1H),3.81(s,2H),3.63(s,3H),3.36(s,2H),2.73(s,1H),1.38(d,J＝6.0Hz,6H).MS(ESI)m/z(M+H)⁺407.4。

Dissolving the product N- (4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazole-3-yl) benzyl) -glycine methyl ester (200mg, 49.24mmol) in 5ml of methanol, adding lithium hydroxide (6mg, 246.2mmol), heating for reaction for 3-5h, cooling, dropwise adding a 1mol/L concentrated hydrochloric acid solution to pH 3, evaporating the solvent, adding distilled water, performing suction filtration, washing and drying to obtain a white solid, namely 115 mg. The yield thereof was found to be 59.3%.¹H NMR(400MHz,DMSO-d₆)δ8.49(d,J＝2.0Hz,1H),8.39(dd,J＝8.8,2.4Hz,1H),8.11(d,J＝8.4Hz,2H),7.70(d,J＝8.4Hz,2H),7.56(d,J＝9.2Hz,1H),4.97(hept,J＝6.0Hz,1H),4.18(s,2H),3.66(s,2H),1.38(d,J＝6.0Hz,6H).MS(ESI)m/z(M+H)⁺393.7。

(6) Preparation of target compound LK-S1P-002

N- (4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazol-3-yl) benzyl) -N-methylglycine hydrochloride

According to the same production method as that of the aforementioned objective compound LK-S1P-001, N- (4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazol-3-yl) benzyl) -N-methylglycine ethyl ester was first produced in the form of a white solid (170 mg) with a yield of 65.4% by replacing glycine methyl ester hydrochloride with sarcosine ethyl ester hydrochloride.¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝2.0Hz,1H),8.38(dd,J＝9.0,1.8Hz,1H),8.03(d,J＝8.0Hz,2H),7.53(t,J＝7.8Hz,3H),4.97(hept,J＝5.8Hz,1H),4.10(q,J＝7.1Hz,2H),3.73(s,2H),3.33(s,2H),2.28(s,3H),1.38(d,J＝6.0Hz,6H),1.20(t,J＝7.0Hz,3H).MS(ESI)m/z(M+H)+421.7。

Further, the same preparation method as that of LK-S1P-001 was employed, and hydrolysis and acidification reactions were performed to obtain LK-S1P-002 as a white solid (102 mg, 55.9% yield).¹H NMR(400MHz,DMSO-d₆)δ8.50(d,J＝2.4Hz,1H),8.40(dd,J＝9.0,2.2Hz,1H),8.16(d,J＝8.4Hz,2H),7.79(d,J＝8.4Hz,2H),7.56(d,J＝9.2Hz,1H),4.98(hept,J＝6.1Hz,1H),4.45(s,2H),4.07(s,2H),2.79(s,3H),1.38(d,J＝6.0Hz,6H).MS(ESI)m/z(M+H)⁺407.7。

(7) Preparation of target Compound LK-S1P-003

(4- (5- (-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) glycinate hydrochloride

In accordance with the foregoing objectsCompound LK-S1P-001 was prepared in the same manner as intermediate I-4-2 by first replacing intermediate I-4-1 with intermediate I-4-2 to give (4- (5- (3-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) glycine methyl ester as a white solid (144 mg, 59.6% yield).¹H NMR(400MHz,DMSO-d₆)δ8.47(d,J＝2.4Hz,1H),8.37(dd,J＝9.0,2.2Hz,1H),8.02-8.00(m,2H),7.54-7.48(m,3H),5.16-5.12(m,1H),3.80(s,2H),3.63(s,3H),3.36(s,2H),2.66(s,1H),1.96-2.05(m,2H),1.62-1.81(m,6H).MS(ESI)m/z(M+H)⁺433.9。

Further, the same preparation method as that of LK-S1P-001 was followed, and hydrolysis and acidification were carried out to obtain LK-S1P-003, which is a white solid (112 mg) with a yield of 70.9%.¹H NMR(400MHz,DMSO-d6)δ10.37(s,1H),8.48(s,1H),8.39(dd,J＝9.2,2.0Hz,1H),8.11(d,J＝8.0Hz,2H),7.70(d,J＝8.0Hz,2H),7.52(d,J＝9.2Hz,1H),5.14-5.17(m,1H),4.19(s,2H),3.69(s,2H),1.97-2.06(m,2H),1.64-1.80(m,6H)；MS(ESI)m/z(M+H)⁺419.2。

(8) Preparation of target compound LK-S1P-004

N- (4- (5- (3-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) -N-methylglycine hydrochloride

According to the same production method as that of the aforementioned objective compound LK-S1P-001, N- (4- (5- (3-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) -N-methylglycine ethyl ester was first produced in the form of a white solid (126 mg) with the intermediate I-4-2 replacing the intermediate I-4-1 and the sarcosine methyl ester hydrochloride replacing the glycine methyl ester hydrochloride.¹H NMR(400MHz,DMSO-d₆)δ8.46(d,J＝2.4Hz,1H),8.38(dd,J＝9.0,2.2Hz,1H),8.04-8.02(m,2H),7.53-7.48(m,3H),5.16-5.12(m,1H),4.10(q,J＝7.2Hz,2H),3.72(s,2H),3.32(s,2H),2.28(s,3H),2.06-1.96(m,2H),1.62-1.82(m,6H),1.20(t,J＝7.2Hz,3H).MS(ESI)m/z(M+H)⁺447.1。

Further preparing the same as LK-S1P-001, hydrolyzing, acidifyingThe objective compound LK-S1P-004 was obtained as a white solid at 111mg in 58.9% yield.¹H NMR(400MHz,DMSO-d6)δ8.45(s,1H),8.37(d,J＝8.0Hz,1H),8.05(d,J＝6.8Hz,2H),7.58(d,J＝6.4Hz,2H),7.50(d,J＝8.4Hz,1H),5.14(s,1H),3.91(s,2H),3.42(s,2H),2.41(s,3H),2.00(m,2H),1.80-1.65(m,6H)；MS(ESI)m/z(M+H)⁺433.6。

(9) Preparation of target Compound LK-S1P-005

1- (4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazol-3-yl) benzyl) pyrrolidine-3-carboxylic acid hydrochloride

Methyl 1- (4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazol-3-yl) benzyl) pyrrolidine-3-carboxylate was first prepared as a white solid in 161mg, yield 61.2%, by the same preparation method as that of the aforementioned objective compound LK-S1P-001, replacing glycine methyl ester hydrochloride with pyrrolidine-3-carboxylate hydrochloride.¹H NMR(400MHz,DMSO-d₆)δ8.48(d,J＝2.0Hz,1H),8.39(dd,J＝8.8,2.4Hz,1H),8.02-8.04(m,2H),7.51-7.56(m,3H),4.97(hept,J＝6.0Hz,1H),3.63-3.70(m,2H),3.61(s,3H),3.02-3.10(m,1H),2.75(t,J＝8.6Hz,1H),2.64-2.68(m,1H),2.53-2.56(m,2H),1.94-2.07(m,2H),1.38(d,J＝6.0Hz,6H).MS(ESI)m/z(M+H)⁺447.1。

Further, the same preparation method as that of LK-S1P-001 was employed, and hydrolysis and acidification were carried out to obtain LK-S1P-005 as a white solid (118 mg) with a yield of 61.4%.¹H NMR(400MHz,DMSO-d6)δ12.04(s,1H),8.50(d,J＝2.4Hz,1H),8.40(dd,J＝9.0,2.2Hz,1H),8.13(d,J＝8.4Hz,2H),7.78(d,J＝8.0Hz,2H),7.56(d,J＝9.2Hz,1H),4.98(hept,J＝6.0Hz,1H),4.34(s,2H),3.26-3.16(m,5H),2.29-2.10(m,2H),1.38(d,J＝6.0Hz,6H)；MS(ESI)m/z(M+H)⁺433.6。

(10) Preparation of target compound LK-S1P-006

1- (4- (5- (3-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) pyrrolidine-3-carboxylic acid hydrochloride

Methyl 1- (4- (5- (3-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) pyrrolidine-3-carboxylate was first prepared as a white solid 144mg in 54.8% yield by the same preparation method of the aforementioned objective compound LK-S1P-001, replacing intermediate I-4-1 with intermediate I-4-2 and replacing glycine methyl ester hydrochloride with pyrrolidine-3-carboxylate hydrochloride.¹H NMR(400MHz,DMSO-d₆)δ8.48(d,J＝2.0Hz,1H),8.39(dd,J＝9.0,2.2Hz,1H),8.03(d,J＝8.1Hz,2H),7.51(d,J＝8.0Hz,3H),5.14-5.17(m,1H),3.66(m,2H),3.60(s,3H),3.02-3.10(m,1H),2.74(t,J＝8.8Hz,1H),2.63-2.67(m,1H),2.52-2.57(m,2H),1.94-2.06(m,4H),1.60-1.81(m,6H)。

Further, the same preparation method as that of LK-S1P-001, hydrolysis and acidification were carried out to obtain the objective compound LK-S1P-006 as a white solid (123 mg, yield 63.7%).¹H NMR(400MHz,DMSO-d6)δ12.94-11.36(m,1H),8.51(d,J＝2.0Hz,1H),8.41(dd,J＝9.0,2.2Hz,1H),8.15(d,J＝8.4Hz,2H),7.82(d,J＝8.0Hz,2H),7.54(d,J＝9.2Hz,1H),5.15-5.18(m,1H),4.46(s,2H),3.35(s,5H),2.16-2.33(m,2H),1.97-2.06(m,2H),1.61-1.82(m,6H)；MS(ESI)m/z(M+H)⁺459.4。

(11) Preparation of target Compound LK-S1P-007

2- (1- (4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazol-3-yl) benzyl) piperidin-4-yl) acetate hydrochloride

Methyl (1- (4- (5- (3-cyano-4-isopropoxyphenyl) -1,2, 4-oxadiazol-3-yl) benzyl) piperidin-4-yl) acetate was first prepared as a white solid in 145mg with the same preparation method as that for the objective compound LK-S1P-001, replacing glycine methyl ester hydrochloride with piperidine methyl acetate hydrochloride.¹H NMR(400MHz,DMSO-d6)δ8.49(d,J＝2.4Hz,1H),8.39(dd,J＝9.0,2.2Hz,1H),8.02-8.04(m,2H),7.55(d,J＝9.2Hz,1H),7.50(d,J＝8.0Hz,1H),4.97(hept,J＝6.1Hz,1H),3.58(s,3H),3.52(s,2H),2.78(d,J＝11.6Hz,2H),2.24(d,J＝6.8Hz,2H),1.93-1.99(m,2H),1.60-1.63(m,3H),1.38(d,J＝6.0Hz,6H),1.17-1.27(m,2H)；MS(ESI)m/z(M+H)⁺475.8。

Further, the same preparation method as that of LK-S1P-001, hydrolysis and acidification were carried out to obtain the objective compound LK-S1P-007 as a white solid (154 mg) with a yield of 673%.¹H NMR(400MHz，DMSO-d6)δ12.24(s,1H),10.78(s,1H),8.50(d,J＝2.3Hz,1H),8.40(dd,J＝9.0,2.3Hz,1H),8.14(d,J＝8.3Hz,2H),7.83(d,J＝8.2Hz,2H),7.57(d,J＝9.3Hz,1H),4.98(p,J＝6.1Hz,1H),4.35(d,J＝4.1Hz,2H),3.31(s,2H),3.04-2.89(m,2H),2.18(d,J＝6.6Hz,2H),1.86(t,J＝15.1Hz,3H),1.58(q,J＝13.0,12.2Hz,2H),1.38(d,J＝6.0Hz,6H)；MS(ESI)m/z(M+H)⁺461.6。

(12) Preparation of target Compound LK-S1P-008

2- (1- (4- (5- (3-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) piperidin-4-yl) acetate hydrochloride

Methyl (1- (4- (5- (3-cyano-4- (cyclopentyloxy) phenyl) -1,2, 4-oxadiazol-3-yl) benzyl) piperidin-4-yl) acetate was first prepared as a white solid at 140mg in 50.8% yield by the same preparation method as the aforementioned objective compound LK-S1P-001, replacing intermediate I-4-1 with intermediate I-4-2 and replacing glycine methyl acetate hydrochloride with piperidine methyl acetate hydrochloride.¹H NMR(400MHz,DMSO-d₆)δ8.48(d,J＝2.0Hz,1H),8.39(dd,J＝9.0,2.2Hz,1H),8.02(d,J＝8.4Hz,2H),7.49-7.52(m,3H),5.13-5.17(m,1H),3.57(s,3H),3.51(s,2H),2.78(d,J＝11.2Hz,2H),2.24(d,J＝6.8Hz,2H),1.92-2.06(m,4H),1.60-1.82(m,9H),1.17-1.25(m,2H).MS(ESI)m/z(M+H)+501.2。

Further, the same preparation method as that of LK-S1P-001 was employed, and hydrolysis and acidification were carried out to obtain LK-S1P-008 as a white solid (143 mg) with a yield of 63.9%.¹H NMR(400MHz,DMSO-d6)δ12.17(s,1H),10.58-10.43(m,1H),8.47(d,J＝2.4Hz,1H),8.37(dd,J＝9.0,2.2Hz,1H),8.12(d,J＝8.0Hz,2H),7.77(d,J＝8.0Hz,2H),7.50(d,J＝9.2Hz,1H),5.11-5.15(m,1H),4.31-4.32(m,2H),3.32(s,2H),2.88-2.98(m,2H),2.15(d,J＝6.8Hz,2H),1.94-2.03(m,2H),1.45-1.83(m,11H)；MS(ESI)m/z(M+H)⁺487.3。

Example 2 exemplary Compound pair of examples of the invention S1P₁And S1P₃In vitro agonistic activity of receptors

Use of³⁵S]GTP γ S binding assay evaluation of the Compound of the invention prepared as above in example 1 on S1P₁And S1P₃In vitro agonistic activity of the receptor. Mu.g of membrane protein (S1P)₁Membrane protein HTS176M, S1P₃Membrane protein HTS097M, Merck Millipore, USA) was mixed with different concentrations of compounds in buffer (20mM HEPES, pH7.4,100mM NaCl,10mM MgCl₂,5μg Saponin,0.5μM GDP，0.3nm[³⁵S]GTP γ S (1200Ci/mmol) was incubated at 30 ℃ for 30 minutes. The reaction was transferred to a deionized water pretreated GF/B filter plate (Millipore MAHFB 1H). The test for radionuclides was performed 3 times with a pre-cooled 10mM sodium phosphate solution (pH7.4), dried and assayed 3 times in parallel for each compound, and the results were averaged as shown in Table 1.

The results show that the compound prepared in example 1 of the present invention is directed to S1P₁The in vitro agonistic activity of the receptor is remarkably superior to that of a positive control drug Ozanimod, particularly the compounds LK-S1P-005 and LK-S1P-006 for S1P₁The in vitro agonistic activity of the receptor is substantially two orders of magnitude higher than Ozanimod; and the compounds have the formula for S1P₃The receptor has no agonism, which shows that the compound of the invention has S1P₁The receptor agonistic strength and selectivity are superior to Ozanimod.

TABLE 1 Pair of Compounds S1P₁And S1P₃In vitro agonistic activity of receptors

Note: "/" indicates in vitro agonist activity >5000 nM.

Example 3 in vivo immunosuppressive experiments of exemplary example compounds of the invention

The immunosuppressive effects of the compounds of the exemplary embodiments of the present invention were evaluated using an index of the number of rat peripheral blood lymphocytes. Healthy male SD rats (weight 200-. Fasted overnight. Blood was collected from the inner canthus vein on the following day and the basic blood lymphocyte count was performed using an ADVIA2120 full-automatic hemocytometer. The test drug was administered to each group at a dose of 1mg/kg, blood was collected from the medial canthus vein 24h after administration, blood lymphocyte count test was performed, percentage of lymphocyte reduction was calculated, and the average of the test results of 3 animals was taken, and the results are shown in table 2.

Table 2 in vivo immunosuppressive action of Compounds

The results show that the in vivo immunosuppressive effect of the compound is generally superior to that of a positive control drug Ozanimod, wherein the compounds LK-S1P-004, LK-S1P-006 and LK-S1P-008 have higher percentage reduction on the number of lymphocytes and show stronger in vivo immunosuppressive activity.

Example 4 in vitro metabolic stability testing of exemplary compounds of the invention

The metabolic stability of exemplary example compounds of the present invention was initially evaluated using a rat liver microsome incubation system evaluation. After incubating the test compound (1. mu.L, 400. mu. mol/L DMSO solution), rat liver microsomal protein (5. mu.L, 20mg/mL) and Tris-HCl buffer (50mM, pH7.4) at 37 ℃ for 5min, 20. mu.L of NADPH (10mM) was added to start the reaction, and the total reaction volume was 200. mu.L (organic solvent content < 0.5%) and the final concentration of the compound was 2. mu. mol/L. After incubation for 0h, 1h, the reaction was stopped with ice acetonitrile (400. mu.L). After vortex mixing, the mixture is centrifuged at 13000g multiplied by 5min, 5 mu L of supernatant is taken for sample injection and LC-MS/MS analysis and detection are carried out. Percent metabolism (%) - (1-Q1h/Q0h) × 100%; q1h/Q0h is the residual amount of the test compound after incubation for 1h/0h, and the results are shown in Table 3.

TABLE 3 in vitro metabolic stability of the Compounds

The result shows that the compound has better in-vitro metabolic stability, more than 85 percent of the drugs keep the original form after 1 hour in a liver microsome incubation system, and the compound is superior to a positive control drug Ozanimod.

EXAMPLE 5 therapeutic Effect of exemplary example Compounds of the invention on Chronic ulcerative colitis

OXZ-induced pathology of chronic ulcerative colitis A part of the active compounds of the invention was evaluated chronically for their therapeutic effect on chronic ulcerative colitis. Male Kunming mice (28-32g) were selected and randomly grouped into 6 mice per group. Mice in the normal control group were abdomen-coated with 0.2ml of 100% ethanol for sensitization, mice in the model group and each administration group were abdomen-coated with 0.2ml of 3% 4-ethoxymethylene-2-phenyloxazoline-5-one (OXZ, E0753) ethanol solution for sensitization, sensitization was repeated one time the next day, after 5 days, 0.15ml of 2% OXZ ethanol (50%) solution was injected from the anus of the mice in the model group and administration group, and 0.15ml of 50% ethanol was injected from the anus in the normal control group. Groups of molded mice were screened on day 3 of the experiment and dosed once daily as follows: (1) normal Control group (Control): directly administering 1% sodium carboxymethylcellulose for intragastric administration. (2) UC Model group (Model): directly administering 1% sodium carboxymethylcellulose for intragastric administration. (3) Positive drug sulfasalazine group (SASP): prepared by 1 percent of sodium carboxymethylcellulose and then perfused into the stomach. (4) Compound group (inventive compound and control compound Ozanimod): 1mg/kg, prepared by 1 percent sodium carboxymethylcellulose and then perfused into the stomach. After 8 days of dosing, groups of animals were sacrificed by cervical dislocation and various indices related to colitis were examined. The average was calculated for 6 of each group and the results are shown in table 4.

TABLE 4 Effect of Compounds on colitis mouse body weight and Colon Length

The result shows that no animal death occurs in the experimental process, the active compound has a certain treatment effect on the chronic ulcerative colitis, and compared with positive drugs SASP and Ozanimod, the active compound has smaller influence on the body weight of a mouse and higher curative effect and safety.

Example 6 evaluation of pharmacokinetic parameters of Compounds of the invention

Healthy male SD rats (weight 200-. (1) Preparing the drug to be detected and the control drug into solution by using normal saline, and performing intragastric administration according to the dosage of 3mg/kg/10 ml; (2) preparing the drug to be tested and the control drug into solution by using normal saline, and performing intravenous injection administration according to the dosage of 0.3mg/kg/5 ml; according to a conventional method for pharmacokinetics research, blood sampling at different time intervals, plasma treatment, standard curve drawing, UPLC-MS/MS analysis on samples and bioavailability calculation are carried out. Pharmacokinetic parameter analysis was performed using Phoenix software. As a result, as shown in Table 5, the compounds of the present invention all had suitable bioavailability.

Table 5 pharmacokinetic non-compartmental model parameters for partial compounds

The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein R is₁Selected from unsubstituted or substituted by one or more R_aSubstituted the following groups: methyl, ethyl, n-propyl, cyclopropyl, n-butyl, isobutyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, cyclopentyl, n-hexyl, isohexyl, cyclohexyl; the R is_aSelected from-F, -Cl, -Br, -I or-CN;

R₂is-CN;

R₃is selected from-NR'₂Or aza C_3-8A cycloalkyl group; wherein, said aza C_3-8Cycloalkyl radicals substituted by one or more-R₆COOR₇Substituted; r' may be the same or different, and is optionally: H. -R₆COOR₇Wherein at least one of R' is-R₆COOR₇(ii) a Wherein R is₆Selected from chemical bonds or C_1-8Alkylene radical, R₇Is selected from H or C_1-8An alkyl group;

2. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 wherein R is₃Is selected from

Wherein R is₄、R₅Is selected from H; l is selected from C_1-6Alkylene, m and n are identical or different and are each independently selected from the group consisting of integers from 1 to 3, R₆Selected from chemical bonds or C_1-8An alkylene group.

3. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 wherein R is₃Selected from-azetidinyl-R₆COOR₇-azacyclopentyl

-R₆COOR₇or-azacyclohexyl-R₆COOR₇。

4. The compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2, wherein the pharmaceutically acceptable salt of the compound is selected from salts of inorganic or organic acids with salifiable sites in the compound;

the inorganic acid is hydrochloric acid, sulfuric acid, hydrobromic acid, phosphoric acid, hydrofluoric acid, hydroiodic acid, pyrosulfuric acid or nitric acid;

the organic acid is citric acid, maleic acid, succinic acid, acetic acid, malic acid, tartaric acid, methanesulfonic acid, benzenesulfonic acid, formic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2- (4-hydroxybenzoyl) benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, p-toluenesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, stearic acid, lactic acid, oxalic acid, malonic acid, adipic acid, alginic acid, fumaric acid, D-gluconic acid, Mandelic acid, ascorbic acid, glucoheptylic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, hemisulfuric acid or thiocyanic acid.

5. The compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 4, wherein the pharmaceutically acceptable salt of the compound is selected from hydrochloride.

6. A compound of formula (I) as claimed in any one of claims 1 to 5, which is a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) is selected from the following compounds or salts thereof:

7. a process for the preparation of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in any one of claims 1 to 6, which comprises the steps of: reacting the compound of formula (I-4) with the compound of formula (I-5) to obtain the compound of formula (I),

wherein R is₁、R₂、R₃Having the definition of any one of claims 1 to 3;

optionally, the preparation method further comprises reacting the compound of formula (I) with a salt-forming reagent to form a pharmaceutically acceptable salt thereof;

optionally, when R₃Group R in (1)₇In the case of alkyl, the resulting compound of formula (I) may be hydrolyzed to yield R₇A compound of formula (I) which is H.

8. The process of claim 7, wherein the compound of formula (I-4) is prepared by a process comprising:

wherein R is₁、R₂、R₃Having the definition of any one of claims 1 to 3;

9. A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds of formula (I) according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof.

10. The pharmaceutical composition of claim 9, further comprising a pharmaceutically acceptable carrier.

11. The pharmaceutical composition of claim 10, wherein the pharmaceutically acceptable carrier is one or more of a filler, a diluent, a disintegrant, a lubricant, a glidant, a binder, a solubilizer, a surfactant, an emulsifier, a preservative, an antioxidant, a flavoring agent, a colorant, an osmotic pressure regulator, a pH regulator, and a proppant.

12. The pharmaceutical composition of claim 9, wherein the amount of the compound of formula (I) or one or more of its pharmaceutically acceptable salts is 0.01-99.99% by weight of the total pharmaceutical composition.

13. A formulation comprising the pharmaceutical composition of claims 10-12.

14. The formulation of claim 13, wherein the formulation is in a liquid, solid, or semi-solid dosage form.

15. The formulation of claim 14, wherein the liquid dosage form is an oral solution formulation, an injection, an infusion solution, a lotion, a drop, a liniment, an aerosol.

16. The formulation of claim 14, wherein the solid dosage form is a tablet, capsule, granule, powder, pill, suppository, patch.

17. The formulation of claim 14, wherein the semisolid dosage form is an ointment, gel, paste.

18. The preparation of claim 13, wherein the dosage form is a general preparation, a sustained release preparation, a controlled release preparation, a targeted preparation, and various microparticle delivery systems.

19. Use of a compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to any one of claims 8 to 12 for the manufacture of a medicament for use in the treatment of S1P₁Use in medicine for a receptor-related disorder.

20. The use of claim 19, wherein S1P₁The receptor-associated disorder is an autoimmune disease, a lymphocyte-mediated disease, an inflammatory disease, a bacterial infection, a fungal infection, a viral infection, or cancer.

21. Use of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to any one of claims 9 to 12 in the manufacture of a medicament for use in the treatment of S1P₁Use in a medicament for the treatment of an inflammatory bowel disease, and/or crohn's disease, and/or ulcerative colitis, and/or systemic lupus erythematosus, and/or rheumatoid arthritis, and/or psoriasis, and/or multiple sclerosis, and/or transplant rejection.