CN112409333A - Phenylpyrrolidines - Google Patents

Abstract

The invention provides a phenylpyrrolidine compound, namely a compound shown as a general formula (A-1) or (A), so as to provide a PDE4 inhibitor which has novel structure, excellent drug effect, high bioavailability and good drug effect, and is used for effectively treating PDE4 related diseases and symptoms, including but not limited to inflammatory diseases, allergic diseases, autoimmune diseases, transplant rejection, joint inflammatory diseases, skin inflammatory diseases, inflammatory bowel diseases, diseases related to smooth muscle contraction and the like.

Description

Phenylpyrrolidines

PRIORITY INFORMATION

The present application claims priority from a chinese patent application having patent application number 201910787733.7, entitled "phenylpyrrolidines" filed on 23/8/2019 with the chinese intellectual property office, and is incorporated herein by reference in its entirety.

Technical Field

The invention relates to a novel phenylpyrrolidine compound, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof, a preparation method thereof, a pharmaceutical composition containing the compound and application of the compound serving as a PDE4 inhibitor.

Background

Phosphodiesterases (abbreviated to PDEs), or more precisely 3', 5' -cyclic nucleotide phosphodiesterases, are enzymes that catalyze the hydrolysis of the second messenger cAMP (cyclic adenosine monophosphate) and cGMP (cyclic guanosine monophosphate) to 5'-AMP (5' -adenosine monophosphate) and 5'-GMP (5' -guanosine monophosphate). Of the 11 phosphodiesterases identified to date, PDE4, PDE7 and PDE8 are specific for cAMP. PDE4 is the most important regulator of cAMP expressed in immune and inflammatory cells such as neutrophils, macrophages and T-lymphocytes (Z. Huang, Current Med Chem,13,2006, p. 3253-3262). Since cAMP is a key second messenger in the regulation of inflammatory responses, PDE4 has been found to regulate the inflammatory response of inflammatory cells by modulating pro-inflammatory cytokines (e.g., TNF α, IL-2, IFN- γ, GM-CSF, and LTB 4). Inhibition of PDE4 is effective in treating inflammatory diseases such as asthma, Chronic Obstructive Pulmonary Disease (COPD), rheumatoid arthritis, atopic dermatitis, inflammatory bowel disease such as Crohn's disease, Atopic Dermatitis (AD), etc. PDE4 includes multiple subtypes such as PDE4A, PDE4B, PDE4C, and PDE4D, and inhibition of PDE4D can cause side effects such as emesis, and selective inhibition of PDE4B can improve the occurrence of emesis side effects (Osamu Suzuki et al, J Pharmacol Sci 123,219-226, 2013).

In addition, cyclic nucleotides (cAMP and cGMP) are important second messengers for regulating smooth muscle contractility. Cyclic nucleotide Phosphodiesterases (PDEs) hydrolyze cyclic nucleotides and are important in regulating the levels of intracellular cyclic nucleotides and the duration of action. Compounds that inhibit PDEs elevate cellular levels of cyclic nucleotides, thereby relaxing various types of smooth muscle. Studies have shown that phosphodiesterase 4(PDE4) specifically hydrolyzes cAMP, and it is abundantly expressed in the bladder. PCT patent application WO2016040083a1 discloses certain azetidinyloxyphenylpyrrolidines which are useful as inhibitors of PDE4 for the treatment of overactive bladder (OAB) and its associated symptoms, such as the relief of frequency and urgency of urination and other disorders.

Despite the advances made in the treatment of inflammatory, allergic, autoimmune and other diseases, there is still a large number of patients in need of better and more effective clinical treatments and regimens.

Disclosure of Invention

The inventors of the present invention have synthesized a novel compound capable of inhibiting the activity of PDE4B and PDE 4D. Therefore, the compound can be used for preparing a PDE4 inhibitor with novel structure, excellent drug effect, high bioavailability and good drug success, and the PDE4 inhibitor is used for inhibiting the activity of PDE4 to up-regulate the cAMP level in immune and inflammatory cells, so that the release of proinflammatory cytokines is reduced, and the inflammatory reaction is further reduced. The compounds provided by the invention are effective in treating diseases and disorders associated with PDE4, including but not limited to inflammatory diseases, allergic diseases, autoimmune diseases, transplant rejection, joint inflammatory diseases, skin inflammatory diseases, inflammatory bowel diseases, diseases associated with smooth muscle contraction, and the like.

The invention provides a compound, which is a compound shown as a general formula (A-1) or a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug of the compound shown as the general formula (A-1):

wherein the content of the first and second substances,

R¹selected from H, unsubstituted or optionally substituted by at least one R^aSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

the at least one R^aEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂Unsubstituted or optionally substituted by at least one R^bSubstituted of the following groups: -NH₂、C₁-C₁₀Alkyl radical, C₁-C₁₀Alkoxy radical, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, C₃-C₁₀Cycloalkyl radical, C₃-C₁₀Cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, C₆-C₂₀Aryl radical, C₆-C₂₀Aryloxy, 5-20 membered heteroaryl, 5-20 membered heteroaryloxy;

the at least one R^bEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂Or the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

R²selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂，

Provided that when R is²When is H, R¹Not methyl, ethyl, isopropyl.

The inventors have searched through a large number of experiments to synthesize a novel compound of the general formula (A-1), and have found that the compound inhibits the activities of the subtypes PDE4D and PDE4B of PDE 4. Since PDE4 specifically hydrolyzes cAMP in cells, the level of cAMP in immune and inflammatory cells can be upregulated by inhibiting the activity of PDE4 using the compounds of the present invention, thereby reducing the release of pro-inflammatory cytokines (e.g., TNF α, IL-2, IFN- γ, GM-CSF, LTB4, etc.), which in turn reduces the inflammatory response. On the other hand, since cAMP is an important second messenger regulating smooth muscle contractility, the level and duration of action of cAMP in cells are regulated by inhibiting the activity of PDE4 using the compounds of the present invention, which in turn relaxes various types of smooth muscle. In view of the above, the compounds of the general formula (A-1) of the present invention are useful for the treatment of diseases and disorders associated with PDE4, including but not limited to inflammatory diseases, allergic diseases, autoimmune diseases, transplant rejection, arthritic diseases, skin inflammatory diseases, inflammatory bowel diseases, and diseases associated with smooth muscle contraction, etc.

The invention provides a compound shown as a general formula (A), and a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof:

wherein R is¹Selected from H, unsubstituted or optionally substituted by one or more R^aSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

each R^aIdentical or different from each other, independently of one another, selected from F, Cl, Br, I, OH, CN, ═ O, NO₂Unsubstituted or optionally substituted by one or more R^bSubstituted of the following groups: -NH₂、C₁-C₁₀Alkyl radical, C₁-C₁₀Alkoxy radical, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkenyloxy radical, C₂-C₁₀Alkynyl, C₂-C₁₀Alkynyloxy, C₃-C₁₀Cycloalkyl radical, C₃-C₁₀Cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, C₆-C₂₀Aryl radical, C₆-C₂₀Aryloxy, 5-20 membered heteroaryl, 5-20 membered heteroaryloxy;

each R^bThe same or different, each is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂Or the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

wherein R is¹Not methyl, ethyl, isopropyl.

According to an embodiment of the present invention, a compound represented by the general formula (a), a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof:

R¹selected from unsubstituted or optionally substituted by one or more R^aSubstituted C₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl; r^aThe same or different, each is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂Or the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl; preferably, R^aIs selected from F; wherein R is¹Not methyl, ethyl, isopropyl.

According to an embodiment of the present invention, in the general formula (A-1) or (A), R¹Selected from unsubstituted or optionally substituted by at least one R^aSubstituted C₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl;

the at least one R^aEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂Or the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl.

According to an embodiment of the present invention, in the general formula (A-1), R²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂。

According to an embodiment of the present invention, in the general formula (A-1), R²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀A cycloalkyl group;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, ═ O.

According to an embodiment of the present invention, in the general formula (A-1), R²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₅Alkyl radical, C₃-C₅A cycloalkyl group;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, ═ O.

According to an embodiment of the present invention, in the general formula (A-1), R²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₅Alkyl radical, C₃-C₅A cycloalkyl group;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, respectively.

In a preferred embodiment of the invention, the compound is selected from one of the following structures:

the novel compounds provided by the present invention, such as the aforementioned 001, 002, 003, 004, 005 and 006, are capable of inhibiting the activity of PDE4, subtypes PDE4B and PDE 4D. In addition, since inhibition of PDE4D causes side effects such as emesis, compounds 001, 003, 004, 005 and 006 can selectively inhibit the activity of PDE4B, and thus effectively ameliorate the occurrence of side effects such as emesis.

The invention also provides a pharmaceutical composition which comprises the compound shown in the formula (A-1) and/or (A) or a stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof.

According to an embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant, such as a pharmaceutically acceptable carrier, diluent or excipient.

The compounds provided by the invention are prepared into medicines in various dosage forms, and the medicines are administered to a subject in a therapeutically effective amount, and after being absorbed by the subject, the medicines can treat or improve diseases related to PDE4 by up-regulating cAMP level in cells and further influencing the release of proinflammatory cytokines and the state of smooth muscles.

According to an embodiment of the invention, the pharmaceutical composition further comprises one or more additional therapeutic agents. Wherein the other therapeutic agents function similarly to the compounds of the invention and are useful for treating diseases associated with PDE 4.

The present invention relates to suitable pharmaceutically acceptable salts of the compounds of formula (A-1) or (A), including but not limited to hydrochloride, hydrobromide, sulphate or bisulphate, phosphate or hydrogenphosphate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, gluconate, methanesulphonate, benzenesulphonate or p-toluenesulphonate. As stated hereinbefore, any reference to a compound of the invention herein includes a pharmaceutically acceptable salt, solvate or combination thereof.

In addition to pharmaceutically acceptable salts of the compounds of the present invention, the present invention also includes other salts. They may serve as intermediates in the purification of compounds or in the preparation of other pharmaceutically acceptable salts or may be used in the identification, characterization or purification of compounds of the invention.

Further, the invention relates to a compound shown in a general formula (A-1) or (A), a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt and a prodrug thereof, and an application of the compound in preparing medicines for treating diseases related to phosphodiesterase-4 (PDE 4).

A preferred embodiment of the present invention, wherein the disease associated with phosphodiesterase-4 (PDE4) is selected from the group consisting of inflammatory diseases, allergic diseases, autoimmune diseases, transplant rejection, inflammatory diseases of the joints, inflammatory diseases of the skin, inflammatory bowel diseases and diseases associated with smooth muscle contraction.

A preferred embodiment of the invention wherein the allergic disease is selected from asthma, chronic bronchitis, chronic obstructive pneumonia, allergic rhinitis, adult respiratory distress syndrome.

A preferred embodiment of the invention, wherein the skin inflammatory disease is selected from the group consisting of atopic dermatitis, psoriasis or urticaria.

A preferred embodiment of the invention, wherein the arthritic disease is selected from rheumatoid arthritis, osteoarthritis, gouty arthritis or spondylitis.

A preferred embodiment of the invention, wherein the inflammatory bowel disease is selected from ulcerative colitis or crohn's disease.

A preferred embodiment of the invention wherein the disorder associated with smooth muscle contractility is selected from the group consisting of overactive bladder and its associated symptoms, such as urinary frequency and urgency.

The invention also relates to a method of treating a disease associated with phosphodiesterase-4 (PDE4) comprising administering to a patient a therapeutically effective dose of a pharmaceutical formulation comprising a compound of the invention or a pharmaceutically acceptable salt thereof.

Definition and description of terms

Unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination are within the scope of the present specification.

The term "pharmaceutically acceptable salts" refers to pharmaceutically acceptable salts of non-toxic acids or bases, including salts of inorganic acids and bases, organic acids and bases. Salts derived from inorganic bases include, but are not limited to, metal salts formed from Al, Ca, Li, Mg, K, Na, and Zn; salts derived from organic bases include, but are not limited to, salts of primary, secondary or tertiary amines, including naturally occurring substituted or unsubstituted amines, cyclic amines, and basic ion exchange resins, such as ammonium, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, caffeine, procaine, choline, betaine, phentermine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, or polyamine resins; salts derived from inorganic and organic acids include, but are not limited to, organic salts formed from sulfuric, phosphoric, nitric, hydrobromic, hydrochloric, formic, acetic, propionic, benzenesulfonic, benzoic, phenylacetic, salicylic, alginic, anthranilic, camphoric, citric, ethenesulfonic, formic, fumaric, furoic, gluconic, glucuronic, glutamic, glycolic, isethionic, lactic, maleic, malic, mandelic, mucic, pamoic, pantothenic, stearic, succinic, sulfanilic, tartaric, p-toluenesulfonic, malonic, 2-hydroxypropionic, oxalic, glycolic, glucuronic, galacturonic, citric, lysine, arginine, aspartic, cinnamic, p-toluenesulfonic, methanesulfonic, ethanesulfonic, or trifluoromethanesulfonic acids and the like.

The term "stereoisomer" refers to isomers resulting from the different arrangement of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers, and conformers.

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in two positions in a molecule. The compounds of the invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer often result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; whereas in phenol the enol type predominates. The present invention encompasses all tautomeric forms of the compounds.

The term "pharmaceutical composition" denotes a mixture of one or more compounds of the invention or physiologically/pharmaceutically acceptable salts or prodrugs thereof with other chemical components, such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration of the compound to an organism, facilitate absorption of the active ingredient and exert biological activity.

The term "solvate" means that the compound of the present invention or a salt thereof includes a stoichiometric or non-stoichiometric amount of solvent bonded with non-covalent intermolecular forces, and when the solvent is water, it is a hydrate.

The term "prodrug" refers to a compound of the invention that can be converted to a biologically active compound under physiological conditions or by solvolysis. Prodrugs of the invention are prepared by modifying functional groups in the compounds, which modifications may be routinely made or removed in vivo to provide the parent compound. Prodrugs include compounds of the present invention wherein a hydroxy or amino group is attached to any group that, when administered to a mammalian subject, cleaves to form a free hydroxy or a free amino group, respectively.

The term "C₁-C₁₀Alkyl "is understood to mean a straight-chain or branched, saturated monovalent hydrocarbon radical having 1,2, 3,4, 5,6, 7, 8, 9 or 10 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2-methylpentyl3-dimethylbutyl, 1, 3-dimethylbutyl, or 1, 2-dimethylbutyl; "C₁-C₆Alkyl "is understood to mean a straight-chain or branched, saturated monovalent hydrocarbon radical having 1,2, 3,4, 5,6 carbon atoms.

The term "C₂-C₁₀Alkynyl "is understood as preferably meaning a straight-chain or branched, monovalent hydrocarbon radical which contains one or more triple bonds and has 2,3, 4,5, 6, 7, 8, 9 or 10 carbon atoms.

The term "C₃-C₁₀Cycloalkyl "is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 10 carbon atoms. Such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group such as a decaline ring. The term "C₃-C₆Cycloalkyl "is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms.

The term "3-10 membered heterocyclyl" means a saturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 1-5, preferably 1-3 heteroatoms selected from N, O and S. 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, dihydroisoquinolinyl. According to the invention, the heterocyclic radical is non-aromatic.

The term "C₆-C₂₀Aryl "is to be understood as preferably meaning a mono-, bi-or tricyclic hydrocarbon ring of monovalent or partially aromatic character having 6 to 20 carbon atoms. In particular a ring having 6 carbon atoms ("C)₆Aryl "), such as phenyl; or a ring having 9 carbon atoms ("C)₉Aryl group), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C)₁₀Aryl 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 "C₆-C₂₀Aryloxy "is defined as above in" C₆-C₂₀Aryl "is linked to at least one oxy" -O- ".

The term "5-20 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: having 5 to 20 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, such as "5-14 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include such monovalent monocyclic, bicyclic or tricyclic aromatic ring systems: which has 5,6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and which comprises 1 to 5, preferably 1 to 3, heteroatoms each independently selected from N, O and S, and in each case additionally can be benzofused. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, thia-4H-pyrazolyl and the like and their benzo derivatives, such as benzofuryl, 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 "5-20 membered heteroaryloxy" means that at least one oxy "-O-" group is attached to the aforementioned "5-20 membered heteroaryl" group.

The term "adjuvant" refers to a pharmaceutically acceptable inert ingredient. Non-limiting examples of the kind of the term "excipient" include binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents and the like. Excipients enhance the handling characteristics of the pharmaceutical formulation, i.e., make the formulation more amenable to direct compression by increasing flowability and/or cohesiveness. Examples of typical "pharmaceutically acceptable carriers" suitable for use in the above formulations are: sugars such as lactose, sucrose, mannitol, and sorbitol; starches, such as corn starch, tapioca starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and methyl cellulose; calcium phosphates such as dicalcium phosphate and tricalcium phosphate; sodium sulfate; calcium sulfate; polyvinylpyrrolidone; polyvinyl alcohol; stearic acid; alkaline earth metal stearates, such as magnesium stearate and calcium stearate; stearic acid; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil and corn oil; nonionic, cationic and anionic surfactants; a glycol polymer; fatty alcohols; and grain hydrolyzed solids and other nontoxic compatible excipients commonly used in pharmaceutical formulations, such as fillers, binders, disintegrants, buffers, preservatives, antioxidants, lubricants, colorants, and the like.

The present invention also relates to a process for the preparation of a compound of formula (a) according to the invention, which process comprises:

wherein R is¹As defined above; x is halogen, such as Cl, Br, I, etc.; the structure of formula (IV) is

Wherein, the compound of formula (I) is in the presence of catalystCarrying out coupling reaction with azetidinyl-3-ol or salt thereof to obtain a compound shown in a formula (II); reacting the compound of the formula (II) with methylsulfonyl chloride to obtain a compound of a formula (III), further carrying out nucleophilic substitution reaction on the compound of the formula (III) and a compound of a formula (IV) to obtain a compound of a formula (V), and removing a protective agent from the compound of the formula (V) under an acidic condition to obtain a target compound (A).

Synthetic routes and preparation methods for formula (IV) refer to patent applications WO2001047905a1, CN106795137A and journal literature Nichols, p.j.; DeMattei, J.A.org.Lett.2006, 8, 1495-1498.

The present invention provides a synthetic route and a preparation method of the compound of the general formula (A-1) similar to the aforementioned method for preparing the compound of the general formula (A), except that the starting materials at the time of synthesis are different, i.e., the above-mentioned compound of the formula (I) is different.

Drawings

The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:

FIG. 1 shows the clinical scores of groups G1, G2, G3 and G4 in a mouse CIA arthritis model experiment;

FIG. 2 shows the AUC inhibition of clinical scores of groups G2, G3 and G4 in a mouse CIA arthritis model experiment.

Detailed Description

The following examples illustrate the technical solutions of the present invention in detail, but the scope of the present invention includes but is not limited thereto.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) and/or Mass Spectrometry (MS). NMR shift in units of 10^-6(ppm). Solvents for NMR measurement were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and an internal standard was Tetramethylsilane (TMS).

Abbreviations of the present invention are defined as follows:

and (2) CuI: cuprous iodide

DCM: methylene dichloride

DMF: n, N-dimethylformamide

DMSO, DMSO: dimethyl sulfoxide

EA: ethyl acetate

Et₃N: triethylamine

N: equivalent concentration, e.g. 1N hydrochloric acid for 1mol/L hydrochloric acid solution

PE: petroleum ether

Unless indicated to the contrary, the compounds exemplified herein are named and numbered using ChemBioDraw Ultra 13.0. Ms: a methanesulfonyl group; LPS: lipopolysaccharide, Lipopolysaccharide (endotoxin); PBMC: peripheral blood mononuclear cells; TNF α: tumor necrosis factor alpha; PBS: phosphate buffer; FBS: fetal bovine serum; ' IC₅₀"half inhibitory concentration" means the concentration at which half of the maximum inhibitory effect is achieved.

The positive control compounds used in the following examples of the present invention have the structural formula

The reference patent WO2014159012A1 discloses a positive control compound which can be used as an inhibitor of PDE4 and can be used for treating diseases related to PDE 4.

LC-MS,M/Z(ESI):486[M+1]。

Example 1: preparation of object Compound 001

(S) -1- ((3S,4S) -4- (3- ((1- (5-Cyclopropoxypyridin-2-yl) azetidin-3-yl) oxy) -4-methoxyphenyl) -3- ((R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) -2, 3-dihydroxypropan-1-one

(S)-1-((3S,4S)-4-(3-((1-(5-cyclopropoxypyridin-2-yl)azetidin-3-yl)oxy)-4-methoxyphenyl)-3-((R)-1-hydroxyethyl)-3-methylpyrrolidin-1-yl)-2,3-dihydroxypropan-1-one

The synthetic route for the target compound 001 is shown below:

the first step is as follows: synthesis of 1- (5-cyclopropoxypyridin-2-yl) azetidinyl-3-ol (001B)

1-(5-cyclopropoxypyridin-2-yl)azetidin-3-ol

2-bromo-5-cyclopropoxypyridine (001A) (1.6g, 7.47mmol) was dissolved in 20mL of DMSO, and azetidinyl-3-alkoxide (2.03g, 18.6mmol), cuprous iodide (284mg, 1.49mmol), L-proline (343mg, 2.98mmol), and potassium carbonate (3.09g, 22.41mmol) were added, heated to 90 deg.C, and stirred under nitrogen overnight. Cooling to room temperature, diluting with water (300mL), extracting with ethyl acetate (100mL × 3), combining the organic phases, washing the organic phase with saturated brine (100mL × 2), separating the liquids, drying the organic phase with anhydrous sodium sulfate, filtering, concentrating, and separating and purifying the residue with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 2:1) to obtain compound 1- (5-cyclopropoxypyridin-2-yl) azetidin-3-ol (001B) as a pale yellow solid (650mg, 42.4% yield).

The second step is that: synthesis of 1- (5-Cyclopropoxypyridin-2-yl) azetidinyl-3-methylsulfonate (001C)

1-(5-cyclopropoxypyridin-2-yl)azetidin-3-yl methanesulfonate

1- (5-Cyclopropoxypyridin-2-yl) azetidin-3-ol (001B) (650mg, 3.16mmol) was dissolved in dichloromethane (15mL), triethylamine (638mg, 6.32mmol) was added, cooling to 0 ℃ and methanesulfonyl chloride (540mg, 4.73mmol) was added and stirring at room temperature for 2 h. Dichloromethane (60mL) was added for dilution, washed with water (60mL × 3), separated, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column separation (petroleum ether: ethyl acetate (V/V) ═ 2:1) to give compound 1- (5-cyclopropoxypyridin-2-yl) azetidinyl-3-methanesulfonate (001C) as a pale yellow solid (460mg, 51.3% yield).

The third step: synthesis of ((3S,4S) -4- (3- ((1- (5-cyclopropoxypyridin-2-yl) azetidinyl) oxy) -4-methoxyphenyl) -3- ((R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) methanone (001D)

((3S,4S)-4-(3-((1-(5-cyclopropoxypyridin-2-yl)azetidin-3-yl)oxy)-4-methoxyphenyl)-3-((R)-1-hydroxyethyl)-3-methylpyrrolidin-1-yl)((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methanone

Compound ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) (3-4-hydroxy-4-methoxyphenyl) - (3- (R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) methanone (compound IV) (380mg, 1mmol) was dissolved in DMF (5mL), potassium phosphate (466mg, 2.50mmol) was added, heated to 90 ℃ and stirred for 0.5h, cooled to 40 ℃,1- (5-cyclopropoxypyridin-2-yl) azetidinyl-3-methanesulfonate (001C) (460mg, 2.50mmol) was added, heated to 90 ℃ and stirred overnight. Cooling to room temperature, diluting with water (60mL), extracting with ethyl acetate (60mL × 3), combining the organic phases, washing the organic phase with saturated brine (60mL × 2), separating the layers, drying the organic phase with anhydrous sodium sulfate, filtering, concentrating, purifying the residue with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 2:1) to give compound ((3S,4S) -4- (3- ((1- (5-cyclopropoxypyridin-2-yl) azetidinyl) oxy) -4-methoxyphenyl) -3- ((R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) methanone (001D), as a white solid (350mg, 61.7% yield).

The fourth step: synthesis of (S) -1- ((3S,4S) -4- (3- ((1- (5-Cyclopropoxypyridin-2-yl) azetidin-3-yl) oxy) -4-methoxyphenyl) -3- ((R) -1-hydroxyethyl) 3-methylpyrrolidin-1-yl) -2, 3-dihydroxypropan-1-one (target Compound 001)

(S)-1-((3S,4S)-4-(3-((1-(5-cyclopropoxypyridin-2-yl)azetidin-3-yl)oxy)-4-methoxyphenyl)-3-((R)-1-hydroxyethyl)-3-methylpyrrolidin-1-yl)-2,3-dihydroxypropan-1-one

((3S,4S) -4- (3- ((1- (5-cyclopropoxypyridin-2-yl) azetidinyl) oxy) -4-methoxyphenyl) -3- ((R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) methanone (001D) (350mg, 0.62mmol) was dissolved in tetrahydrofuran (10mL), 1N hydrochloric acid (8mL) was added, and the mixture was stirred at room temperature overnight. Saturated aqueous sodium bicarbonate solution was added, pH was adjusted to 8 to 9, extraction was performed with ethyl acetate (25mL × 3), organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (pure ethyl acetate) to obtain compound (S) -1- ((3S,4S) -4- (3- ((1- (5-cyclopropoxypyridin-2-yl) azetidin-3-yl) oxy) -4-methoxyphenyl) -3- ((R) -1-hydroxyethyl) 3-methylpyrrolidin-1-yl) -2, 3-dihydroxypropan-1-one (target compound 001) as a white solid (230mg, 70.6%).

¹H NMR(400MHz,DMSO-d6)δ7.86-7.85(m,1H),7.80-7.77(m,1H),7.70-6.97(dd,1H),6.93-6.88(m,2H),6.72-6.71(m,1H),6.47(s,3H),5.19-5.16(m,1H),4.62-4.59(m,2H),4.30-4.24(m,1H),4.21-4.14(m,2H),3.97-3.94(m,2H),3.77(s,3H),3.74-3.45(m,6H),3.39-3.34(m,1H),1.04(d,3H),0.82-0.78(m,2H),0.71-0.68(m,2H),0.66(d,3H).

LC-MS,M/Z(ESI):528.2(M+1).

Example 2: preparation of target Compound 002

(S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one

(S)-2,3-dihydroxy-1-((3S,4S)-3-((R)-1-hydroxyethyl)-4-(4-methoxy-3-((1-(5-(trifluorometh oxy)pyridin-2-yl)azetidin-3-yl)oxy)phenyl)-3-methylpyrrolidin-1-yl)propan-1-one

The target compound 002 was prepared by the preparation method of reference example 1 using 2-bromo-5- (trifluoromethoxy) pyridine (002A) as a starting material.

¹H NMR(400MHz,DMSO-d6)δ8.14(s,1H),7.62(d,1H),6.93(dd,1H),6.89(d,1H),6.69(d,1H),6.54(d,1H),5.17-5.10(m,1H),4.88(s,2H),4.77(s,1H),4.42(dd,2H),4.24(d,1H),3.97-3.91(m,3H),3.75(s,3H),3.74-3.45(m,6H),1.02(d,3H),0.64(d,3H).

LC-MS,M/Z(ESI):556.2(M+1).

Example 3: preparation of target Compound 003

(S) -1- ((3S,4S) -4- (3- ((1- (5-cyclobutoxypyridin-2-yl) azetidin-3-yl) oxy) -4-methoxyphenyl) -3- ((R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) -2, 3-dihydroxypropan-1-one

(S)-1-((3S,4S)-4-(3-((1-(5-cyclobutoxypyridin-2-yl)azetidin-3-yl)oxy)-4-methoxyphenyl)-3-((R)-1-hydroxyethyl)-3-methylpyrrolidin-1-yl)-2,3-dihydroxypropan-1-one

The objective compound 003 was obtained by the preparation method of reference example 1 using 2-bromo-5-cyclobutyloxypyridine (003A) as a starting material.

¹H NMR(400MHz,DMSO-d6)δ7.75(d,1H),7.20-7.16(m,1H),6.95(dd,1H),6.87-6.85(m,1H),6.68(s,1H),6.44(d,1H),5.10-5.08(m,1H),4.92-4.83(m,2H),4.76-4.71(m,1H),4.61-4.55(m,1H),4.31-4.21(m,3H),3.95-3.91(m,1H),3.82-3.80(m,2H),3.75(s,3H),3.74-3.45(m,6H),2.51-2.33(m,3H),2.02-1.97(m,2H),1.76-1.74(m,1H),1.65-1.56(m,1H),1.02(d,3H),0.64(d,3H).

LC-MS,M/Z(ESI):542.2(M+1).

Example 4: preparation of target Compound 004

(S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (5- (oxetanyl-3-yloxy) pyridin-2-yl) azetidinyl-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one

(S)-2,3-dihydroxy-1-((3S,4S)-3-((R)-1-hydroxyethyl)-4-(4-methoxy-3-((1-(5-(oxetan-3-yloxy)pyridin-2-yl)azetidin-3-yl)oxy)phenyl)-3-methylpyrrolidin-1-yl)propan-1-one

The objective compound 004 was prepared by the preparation method of reference example 1 using 2-bromo-5- (oxetan-3-yloxy) pyridine (004A) as a starting material.

¹H NMR(400MHz,DMSO-d6)δ7.70(d,1H),7.19-7.16(m,1H),6.93(dd,1H),6.87-6.85(m,1H),6.68(s,1H),6.46(d,1H),5.23-5.17(m,1H),5.11-5.08(m,1H),4.92-4.85(m,4H),4.76-4.71(m,1H),4.54-4.51(m,2H),4.32-4.23(m,3H),3.93-3.91(m,1H),3.84-3.78(m,2H),3.75(s,3H),3.68-3.45(m,6H),3.19-3.16(m,1H),1.01(d,3H),0.64(d,3H).

LC-MS,M/Z(ESI):544.3(M+1).

Example 5: preparation of target Compound 005

(S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one

(S)-2,3-dihydroxy-1-((3S,4S)-3-((R)-1-hydroxyethyl)-4-(4-methoxy-3-((1-(6-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-yl)oxy)phenyl)-3-methylpyrrolidin-1-yl)propan-1-one

The first step is as follows: synthesis of O- (6-bromo-2-methylpyridin-3-yl) thiocarbonyl chloride (005B)

O-(6-bromo-2-methylpyridin-3-yl)carbonochloridothioate(005B)

2-bromo-5-hydroxy-6-methylpyridine (005A) (30.0g, 160.4mmol) was added to 210mL of a 5% NaOH solution at 0 deg.C, carbon dichlorosulfide (20.0g, 173.9mmol) in 120mL of chloroform was added, and the mixture was stirred at 0 deg.C for 2 h. After warming to room temperature, extraction was performed with chloroform (120 mL. times.3), the organic phases were combined, washed with saturated brine (50 mL. times.2), separated, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound as a crude pale yellow liquid O- (6-bromo-2-methylpyridin-3-yl) thiocarbonyl chloride (005B) (45.2g crude).

The second step is that: synthesis of O- (6-bromo-2-methylpyridin-3-yl) methylmercaptothiocarbonate (005C)

O-(6-bromo-2-methylpyridin-3-yl)S-methyl carbonodithioate

O- (6-bromo-2-methylpyridin-3-yl) thiocarbonyl chloride (005B) (45.2g,160.4mmol) was added to 200mL of chloroform at room temperature, sodium thiomethoxide (12.4g,177.1mmol) was added, and the mixture was stirred at room temperature for 72 hours. Water (200mL) was added for dilution, extraction was performed with chloroform (120mL × 3), the organic phases were combined, the organic phase was washed with saturated brine (50mL × 2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (PE: EA ═ 10:1) to give the title compound O- (6-bromo-2-methylpyridin-3-yl) methylmercaptothiocarbonate (005C) as a pale yellow liquid (15.8g, yield in two steps 35.6%).

The third step: synthesis of 6-bromo-2-methyl-3- (trifluoromethoxy) pyridine (005D)

6-bromo-2-methyl-3-(trifluoromethoxy)pyridine(005D)

Dibromohydantoin (50.5g,176.4mmol) was added to 300mL DCM at room temperature, cooled to-78 deg.C,70% pyridine hydrofluoric acid solution (400mL,3136.8mmol) was added at low temperature and stirred for 45 min. A solution of O- (6-bromo-2-methylpyridin-3-yl) methylthio-carbonate (005C) (10.9g,39.2mmol) in 100mL of DCM was slowly added dropwise, after dropping, the temperature was slowly raised to-5 ℃ and the mixture was stirred for 1.5 hours. The reaction was poured into 500mL of cold saturated NaHCO₃To the solution, pH was adjusted to 10 to 12 with NaOH, extraction was performed with DCM (200mL × 3), organic phases were combined, the organic phase was washed with saturated brine (200mL × 2), liquid separation was performed, the organic phase was dried over anhydrous sodium sulfate, filtration and concentration were performed, and the residue was separated and purified with a silica gel column (PE: EA ═ 10:1) to obtain the title compound 6-bromo-2-methyl-3- (trifluoromethoxy) pyridine (005D) (7.0g, yield 69.7%) as a pale yellow liquid.

LC-MS,M/Z(ESI):256.3(M+1).

The fourth step: synthesis of 1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-ol (005E)

1-(6-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-ol(005E)

6-bromo-2-methyl-3- (trifluoromethoxy) pyridine (005D) (2.56g,10.0mmol) was added to 30mL of DMSO at room temperature, azetidinyl-3-alkoxide (1.65g,15.0mmol), cuprous iodide (190mg,1.0mmol), L-proline (1.27g,11.0mmol), and potassium phosphate (4.24g,20.0mmol) were added, heated to 110 deg.C, and stirred overnight. Cooled to room temperature, diluted with water (150mL), extracted with ethyl acetate (50mL × 3), the organic phases combined, washed with saturated brine (50mL × 2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue purified by silica gel column separation (PE: EA ═ 2:1) to give the title compound 1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-ol (005E) (1.0g, 40.3% yield) as a pale yellow solid.

LC-MS,M/Z(ESI):249.4(M+1).

The fifth step: synthesis of 1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl methanesulfonate (005F)

1-(6-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-yl methanesulfonate(005F)

1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-ol (005E) (140mg,0.56mmol) was added to 3mL of dichloromethane, triethylamine (171mg,1.69mmol) was added, methanesulfonyl chloride (97mg,0.85mmol) was added and stirred at room temperature for 2 h. Dichloromethane (10mL) was added for dilution, washed with water (10mL × 3), separated, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column separation (PE: EA ═ 2:1) to give the title compound 1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-ylmethanesulfonate (005F) as a pale yellow solid (150mg, 81.5% yield).

LC-MS,M/Z(ESI):327.2(M+1).

And a sixth step: synthesis of ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) methanone (005G)

((S)-2,2-dimethyl-1,3-dioxolan-4-yl)((3S,4S)-3-((R)-1-hydroxyethyl)-4-(4-methoxy-3-((1-(6-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-yl)oxy)phenyl)-3-methylpyrrolidin-1-yl)methanone(005G)

The compound ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) (3-hydroxy-4-methoxyphenyl) - (3- (R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) methanone (1.30g,3.37mmol) was added to DMF (20mL), potassium phosphate (1.40g,6.74mmol) was added, 1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-methanesulfonate (005F) (1.10g,3.37mmol) was added, heated to 110 ℃ and stirred overnight. Cooled to room temperature, diluted with water (60mL), extracted with ethyl acetate (60mL × 3), the organic phases combined, washed with saturated brine (60mL × 2), separated, the organic phase dried over anhydrous sodium sulfate, filtered, concentrated and the residue purified on a silica gel column (PE: EA ═ 2:1) to give the title compound ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) methanone (005G) (1.40G, yield 68.1%).

LC-MS,M/Z(ESI):610.3(M+1).

The seventh step: synthesis of (S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one (005)

(S)-2,3-dihydroxy-1-((3S,4S)-3-((R)-1-hydroxyethyl)-4-(4-methoxy-3-((1-(6-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-yl)oxy)phenyl)-3-methylpyrrolidin-1-yl)propan-1-one(005)

((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) methanone (005G) (400mg,0.66mmol) was dissolved in tetrahydrofuran (10mL), 2N hydrochloric acid (2mL) was added, and stirring was carried out at room temperature overnight. Adding saturated aqueous sodium bicarbonate solution, adjusting the pH to 8-9, extracting with ethyl acetate (25 mL. times.3), combining the organic phases, drying with anhydrous sodium sulfate, filtering, concentrating, separating and purifying the residue with silica gel column (pure EA), the title compound was obtained as a white solid (S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (6-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one (005) (340mg, 90.9% yield).

¹H NMR(400MHz,DMSO-d6)δ7.59(d,1H),6.96(dd,1H),6.88(d,1H),6.68(s,1H),6.37(d,1H),5.12(d,1H),4.92(b,2H),4.41(t,2H),4.26-4.22(m,1H),3.94-3.91(m,3H),3.75(s,3H),3.74-3.48(m,8H),2.31(s,3H),1.02(d,3H),0.64(s,3H).

LC-MS,M/Z(ESI):570.3(M+1).

Example 6: preparation of target compound 006

(S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one

(S)-2,3-dihydroxy-1-((3S,4S)-3-((R)-1-hydroxyethyl)-4-(4-methoxy-3-((1-(4-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-yl)oxy)phenyl)-3-methylpyrrolidin-1-yl)propan-1-one

The first step is as follows: synthesis of O- (6-bromo-4-methylpyridin-3-yl) thiocarbonyl chloride (006B)

O-(6-bromo-4-methylpyridin-3-yl)carbonochloridothioate(006B)

6-bromo-3-hydroxy-4-methylpyridine (006A) (13.5g,71.8mmol) was added to 95mL of a 5% NaOH solution at 0 deg.C, followed by addition of carbon dichlorosulfide (9.1g,173.9mmol) in 55mL of chloroform and stirring at 0 deg.C for 2 h. After warming to room temperature, extraction was performed with chloroform (120 mL. times.3), the organic phases were combined, washed with saturated brine (50 mL. times.2), separated, dried over anhydrous sodium sulfate, filtered, and concentrated to give the title compound crude O- (6-bromo-2-methylpyridin-3-yl) thiocarbonyl chloride (006B) as a pale yellow liquid (20.0g crude was used directly in the next step).

The second step is that: synthesis of O- (6-bromo-4-methylpyridin-3-yl) methylmercaptothiocarbonate (006C)

O-(6-bromo-4-methylpyridin-3-yl)S-methyl carbonodithioate(006C)

Crude O- (6-bromo-4-methylpyridin-3-yl) thiocarbonyl chloride (006B) (20.0g,71.8mmol) from the previous step was added to 100mL of chloroform at room temperature, sodium thiomethoxide (5.5g,78.9mmol) was added, and the mixture was stirred at room temperature for 72 h. Water (100mL) was added for dilution, extraction was performed with chloroform (50mL × 3), the organic phases were combined, the organic phase was washed with saturated brine (50mL × 2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (PE: EA ═ 10:1) to give the title compound O- (6-bromo-4-methylpyridin-3-yl) methylmercaptothiocarbonate (006C) as a pale yellow liquid (10.0g, 50.1% yield in two steps).

The third step: synthesis of 6-bromo-4-methyl-3- (trifluoromethoxy) pyridine (006D)

6-bromo-4-methyl-3-(trifluoromethoxy)pyridine(006D)

Dibromohydantoin (37.0g,129.5mmol) was added to 300mL DCM at room temperature, cooled to-78 deg.C, added 60% pyridine hydrofluoric acid solution (368mL,2473.6mmol) at low temperature, and stirred for 45 min. A solution of O- (6-bromo-4-methylpyridin-3-yl) S-methyldithiocararbonate (006C) (8.0g,28.7mmol) in 100mL of DCM was added dropwise slowly, after dropping, the temperature was raised slowly to-5 ℃ and the mixture was stirred for 1.5 h. The reaction was poured into 500mL of cold saturated NaHCO₃To the solution, pH was adjusted to 10 to 12 with NaOH, extraction was performed with DCM (200mL × 3), organic phases were combined, the organic phase was washed with saturated brine (200mL × 2), liquid separation was performed, the organic phase was dried over anhydrous sodium sulfate, filtration and concentration were performed, and the residue was separated and purified by a silica gel column (PE: EA ═ 10:1) to obtain the title compound 6-bromo-4-methyl-3- (trifluoromethoxy) pyridine (006D) (6.0g, yield 81.9%) as a pale yellow liquid.

LC-MS,M/Z(ESI):256.3(M+1).

The fourth step: synthesis of 1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-ol (006E)

1-(4-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-ol(006E)

6-bromo-4-methyl-3- (trifluoromethoxy) pyridine (006D) (6.0g,23.4mmol) was added to 65mL of DMSO at room temperature, azetidinyl-3-alkoxide (3.6g,32.7mmol), cuprous iodide (0.50g,2.63mmol), L-proline (3.0g,26.1mmol), and potassium phosphate (10.0g,47.1mmol) were added, and the mixture was heated to 110 ℃ and stirred overnight. Cooled to room temperature, diluted with water (150mL), extracted with ethyl acetate (50mL × 3), the organic phases combined, washed with saturated brine (50mL × 2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue purified by silica gel column separation (PE: EA ═ 2:1) to give the title compound 1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-ol (006E) (4.6g, 79.1% yield) as a pale yellow solid.

LC-MS,M/Z(ESI):249.4(M+1).

The fifth step: synthesis of 1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl methanesulfonate (006F)

1-(4-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-yl methanesulfonate(006F)

1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-ol (006E) (600mg,2.42mmol) was added to 8mL of dichloromethane, triethylamine (733mg,7.26mmol) was added, methanesulfonyl chloride (413mg,3.63mmol) was added and the mixture was stirred at room temperature for 4 h. Dichloromethane (20mL) was added for dilution, washed with water (10mL × 3), separated, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column separation (PE: EA ═ 2:1) to give the title compound 1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-ylmethanesulfonate (006F) as a pale yellow solid (750mg, 95.0% yield).

LC-MS,M/Z(ESI):327.2(M+1).

And a sixth step: synthesis of ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) methanone (006G)

((S)-2,2-dimethyl-1,3-dioxolan-4-yl)((3S,4S)-3-((R)-1-hydroxyethyl)-4-(4-methoxy-3-((1-(4-methyl-5-(trifluoromethoxy)pyridin-2-yl)azetidin-3-yl)oxy)phenyl)-3-methylpyrrolidin-1-yl)methanone(006G)

The compound ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) (3-hydroxy-4-methoxyphenyl) - (3- (R) -1-hydroxyethyl) -3-methylpyrrolidin-1-yl) methanone (1.0g,2.64mmol) was added to DMF (20mL), potassium phosphate (3.40g,16.0mmol) was added, 1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidinyl-3-methanesulfonate (006F) (0.86g,2.64mmol) was added, heated to 110 ℃ and stirred overnight. Cooled to room temperature, diluted with water (60mL), extracted with ethyl acetate (60mL × 3), the organic phases combined, washed with saturated brine (60mL × 2), separated, the organic phase dried over anhydrous sodium sulfate, filtered, concentrated, and the residue purified on a silica gel column (PE: EA ═ 2:1) to give the title compound (pale yellow solid ((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) methanone (006G) (1.05G, yield 65.3%).

LC-MS,M/Z(ESI):610.3(M+1).

The seventh step: synthesis of (S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one (006)

Synthesis of (S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyi) -4- (4-methoxy-3- ((1- (4-methyl-5- (trifluoromethyl) pyridine-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one (006)

((S) -2, 2-dimethyl-1, 3-dioxolan-4-yl) ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) methanone (006G) (100mg,0.16mmol) was dissolved in tetrahydrofuran (2mL), 2N hydrochloric acid (1mL) was added and stirred at room temperature overnight. Adding saturated aqueous sodium bicarbonate solution, adjusting the pH to 8-9, extracting with ethyl acetate (5mL × 3), combining the organic phases, drying with anhydrous sodium sulfate, filtering, concentrating, separating and purifying the residue with silica gel column (pure EA), the title compound was obtained as a white solid (S) -2,3-dihydroxy-1- ((3S,4S) -3- ((R) -1-hydroxyethyl) -4- (4-methoxy-3- ((1- (4-methyl-5- (trifluoromethoxy) pyridin-2-yl) azetidin-3-yl) oxy) phenyl) -3-methylpyrrolidin-1-yl) propan-1-one (006) (73mg, 78.1% yield).

¹H NMR(400MHz,DMSO-d6)δ8.04(s,1H),6.96(dd,1H),6.89(d,1H),6.69(s,1H),6.46(s,1H),5.13(d,1H),4.92-4.84(m,2H),4.77-4.72(m,1H),4.41(t,2H),4.27-4.25(m,1H),3.94-3.91(m,3H),3.76(s,3H),3.74-3.48(m,7H),2.22(s,3H),1.02(d,3H),0.64(s,3H).

LC-MS,M/Z(ESI):570.3(M+1)

Examples of biological Activity and related Properties

Example 7: PDE4B and PDE4D enzyme activity assays

The inhibitory activity of the positive control compound and the compounds 001-006 provided by the invention on PDE4B and PDE4D can be detected by using PDE-glophosodiesterase Assay Kit (promega, V1361). Briefly, the compounds to be tested are first formulated in DMSO solvent as a 10mM concentrated stock solution, and then diluted into 10 × working solution with the Reaction buffer provided in the kit. Working on ice, PDE4B enzyme (Enzo Life Sciences, BML-SE522-0020) was diluted with Reaction buffer to a concentration of 1 ng/. mu.L, PDE4D enzyme (Enzo Life Sciences)nce, BML-SE523-0020) to a concentration of 4 ng/. mu.L. 1.5ul of PDE4B or PDE4D working solution and 1 ul of compound working solution were added to wells of a 384 well plate (Corning, CLS3707) and incubated with shaking at room temperature for 5 minutes, followed by addition of 2.5 ul/well cAMP (2 uM in Reaction Buffer), continued incubation with shaking at room temperature for 20 minutes, addition of 2.5 ul/well 1 × Termination Buffer, followed by addition of 2.5 ul/well 1 × protection Buffer, and continued shaking at room temperature for 20 minutes. Finally, 10. mu.L/well of 1 XKinase-Glo was added, incubated with shaking at room temperature for 10 minutes, and bioluminescence was detected using a Phearsar instrument. Inputting the experimental result into GraphPad Prism software, and obtaining the IC of each compound through fitting calculation₅₀。

Table 1 shows the inhibitory activity of the positive control group compound and the compounds 001-006 provided by the invention on PDE4B and PDE4D, which are determined by an enzyme activity experiment.

TABLE 1 results of the inhibitory Activity of the test Compounds on PDE4B and PDE4D

Experimental results show that compared with a positive control group compound, the compound 001/003-006 obviously shows better selective inhibition effect on PDE4B, and compared with a positive control group compound, the compound 002 has better inhibition activity on PDE4B and PDE 4D. Thus, the compounds of the present invention are useful as PDE4 inhibitors. In addition, since compounds 001/003 to 006 can selectively inhibit the activity of PDE4B, they are effective in ameliorating the occurrence of side effects such as emesis when used in the treatment of diseases associated with PED 4.

Example 8: TNF alpha secretion model test by LPS-induced human PBMC

And (3) PBMC extraction process: collecting fresh human peripheral concentrated blood, sucking 1 unit of human peripheral concentrated blood (concentrated from 200cc of peripheral blood), quantitatively adding 0.9% physiological saline to total volume of 120ml, and mixing. Taking 50ml centrifuge tube, adding 15ml Lymphoprep respectively^TMThe centrifugal tube is held by hand, the angle of inclination is about 45 degrees, 30ml of diluted concentrated blood is sucked, the concentrated blood is carefully and slowly added in a wall-adhering manner, the diluted blood is overlapped on the layering liquid, and the diluted blood is prevented from being mixedAnd putting the separation liquid into the separation liquid or breaking the liquid level of the separation liquid. Lymphoprep^TMThe ratio to diluted blood was 1: 2. The tube was trimmed and placed in a horizontal centrifuge (eppendorf, 5810R), centrifuged at 800g for 20min at 20 ℃ with an increasing speed of 1 and a decreasing speed of 0. The tube was carefully removed. Directly putting the Pasteur pipette into the leucocyte layer, and sucking PBMC. Adding 3 times of 0.9% physiological saline or PBS (without calcium and magnesium), and gently blowing, beating and mixing. After mixing, the mixture was centrifuged at 20 ℃ and 250g for 10min to remove the remaining platelets in the cell suspension, the supernatant was removed, the cell pellet was suspended in 20ml PBS and counted by trypan blue staining.

PBMC screening process: PBMCs from step 1 were centrifuged to remove PBS and then counted in a complete medium (RPMI1640+ 10% FBS + 1% P/S) resuspended. According to 5X 10^4/Wells, 100 μ L/well, were plated into cells. The compound to be screened was formulated to a final concentration of 4 x. Add 50. mu.L/well to the cells. Preincubation was performed for 30min in advance. Control wells were also set without compound. LPS stimulation was 10ng/ml, 4X, added to the cells at 50. mu.l/well. Control wells were also set, without LPS wells. Cells were incubated further and 10% of the supernatant was collected at 24h for detection. The collected supernatants were assayed according to the Human TNF α kit from Invitrogen (REF: 88-7346-88).

The inhibitory activity of the positive control compound and the compounds 001-006 provided by the invention on TNF alpha secretion of human PBMC induced by LPS is determined according to the method.

TABLE 2 results of inhibitory Activity of test Compounds on TNF α secretion from LPS-induced human PBMCs

Test compounds	IC50(nM)
		Positive control group	3.32
001	0.96
		002	1.45
003	1.67
		004	5.32
005	0.63
		006	0.50

Experimental results show that compared with a positive control group, the compounds 001-003, 005 and 006 of the invention have better TNF alpha secretion inhibition activity on human PBMC, can better inhibit the secretion of inflammatory factors TNF alpha in human PBMC, and have more prominent anti-inflammatory effect. The compound 004 has the TNF alpha secretion inhibiting activity of human PBMC equivalent to that of a positive control group, can also inhibit the secretion of inflammatory factor TNF alpha in human PBMC, and has an anti-inflammatory effect.

Example 9: measurement of TNF alpha Release in LPS-stimulated mice

Taking 18 Balb/c mice, week old: and (3) randomly distributing the medicines into a blank group, a model group and an administration group after 6-8 weeks, and orally intragastrically administering a solvent (the blank group and the model group) or 50mg/kg of the compound (the administration group, wherein the administration dose of the positive control group is 100 mg/kg). 30 minutes after the gavage, PBS (blank group) or 1mg/kg LPS (model group and administration group) was intraperitoneally injected. Collecting blood from mouse heart 90 min or 120 min after intraperitoneal injection, standing at 2-8 deg.C for 4 hr, centrifuging at 5000rpm for 10min, collecting serum, and storing at-80 deg.C.

TNF alpha levels in serum were measured using a Mouse TNF alpha assay kit (Mouse TNFa ELISA kit: Biolegend, Cat: 430904). The release inhibitory activity of the compounds on TNF α was calculated from serum TNF α levels.

Compound inhibitory activity% {1- (administration group TNF α concentration-blank TNF α concentration)/(model group TNF α concentration-blank TNF α concentration) } × 100

The effect of the positive control compound, the compounds 001-003 and 005 and 006 provided by the invention on the release of TNF alpha in mice stimulated by LPS was determined according to the above experimental method.

TABLE 3 determination of TNF α Release in LPS stimulated mice

The experimental result shows that compared with the compounds in the positive control group, the compounds 001-003, 005 and 006 can better inhibit the release of TNF alpha stimulated by LPS in a mouse body, the inhibition activity is obviously superior to that of the positive control group, and the compounds have more prominent anti-inflammatory effect.

Example 10: mouse pharmacokinetic testing

Mouse pharmacokinetic experiments, male CD-1 mice 9, 20-25g, were fasted overnight and given 10mg/kg orally by gavage. 3 mice were bled at each blood collection time point, for a total of 9 mice, alternately before and 15, 30 minutes and 1,2, 4, 8, 24 hours after dosing. Blood samples were centrifuged at 8000 rpm for 6 minutes at 4 ℃ and plasma was collected and stored at-20 ℃. Respectively taking plasma at each time point, adding 3-5 times of acetonitrile solution containing an internal standard, mixing for 1 minute by vortex, centrifuging for 10 minutes at 4 ℃ at 13000 r/min, taking supernate, adding 3 times of water, mixing, taking a proper amount of mixed solution, and carrying out LC-MS/MS analysis. The major pharmacokinetic parameters were analyzed using the WinNonlin 7.0 software non-compartmental model.

TABLE 4 pharmacokinetic test results in mice

The experimental result shows that compared with the compound in the positive control group, the compound 002 of the invention shows more excellent plasma exposure, the Cmax and the AUC0-t are both superior to the positive control group and are about 3 times of the positive control group, and the compound of the invention has excellent pharmacokinetic property.

Example 11: canine pharmacokinetic testing

Dog pharmacokinetic experiments, male beagle dogs were used 3, 8-10kg, fasted overnight, and orally gavaged 5 mg/kg. Blood was collected before dosing and at 15, 30 minutes and 1,2, 4, 8, 24 hours post-dosing. Blood samples were centrifuged at 8000 rpm for 6 minutes at 4 ℃ and plasma was collected and stored at-20 ℃. And (3) adding 3-5 times of acetonitrile solution containing an internal standard into the plasma at each time point, mixing, carrying out vortex mixing for 1 minute, centrifuging at 4 ℃ for 10 minutes at 13000 rpm, taking supernatant, adding 3 times of water, mixing, and taking a proper amount of mixed solution to carry out LC-MS/MS analysis. The major pharmacokinetic parameters were analyzed using the WinNonlin 7.0 software non-compartmental model.

TABLE 5 results of pharmacokinetic experiments in dogs

The experimental result shows that compared with the compound in the positive control group, the compound 001 and the compound 002 of the invention show more excellent plasma exposure, the Cmax and the AUC0-t are both superior to the positive control group, the Cmax is about 2 times of the positive control group, and the exposure AUC0-t is about 3 times of the positive control group; the compounds of the present invention have excellent pharmacokinetic properties.

Example 12: mouse CIA arthritis model test

1. Experimental methods

(1) Preparation of type II collagen/complete Freund's adjuvant

Preparing acetic acid: 2N acetic acid was diluted to 100mM, filtered through a 0.22 μm filter, and stored at 4 ℃.

Preparing a bovine II type collagen solution: bovine type II Collagen (CII) was dissolved in 100mM acetic acid and stored overnight at 4 ℃. The final concentration of collagen was 8 mg/mL.

Preparation of the emulsion: the overnight stored CII solution was mixed with an equal volume of complete freund's adjuvant and homogenized on ice at 30,000 rpm for approximately 60 minutes using a high speed homogenizer until the solution formed a stable emulsion.

(2) Induction of arthritis:

DBA/1 mice were anesthetized with isoflurane and immunized subcutaneously caudally (2-3 cm from the root of the tail) by injection of 50 microliters of the prepared collagen emulsion (containing 200 micrograms of CII). The day of the first immunization was noted as day 0, with subsequent days noted in order. On day 21, the tail was injected with the same volume of collagen emulsion. The normal group of mice did not need to be immunized.

(3) Administration and dose design

On day 21, arthritis-induced mice were randomly grouped by weight to make the average body weight among groups consistent, and randomly divided into 4 treatment groups, each of which was administered with 10 mice.

G1 was a normal mouse, without any treatment; group G2 was administered a blank vehicle; the compounds (positive control compound and compound 002) were administered in the G3 and G4 groups, respectively, at a dose of 30mg/kg 2 times per day for 21 days. The volume of the gavage administration is 10 mL/kg.

(4) Determination of arthritis onset index

After boosting the immunity, mice were observed daily for morbidity. When mice started to develop disease (clinical signs of arthritis appeared), clinical scores were scored according to the different degrees of lesions (redness, joint deformity) on a scale of 0-4 points, with a maximum score of 4 for each limb and 16 for each animal. Scores were scored at least three times per week.

The area under the clinical score curve (AUC) was calculated for each group from 21 days to 42 days, and the percent inhibition of the clinical score AUC was calculated for the dosing group:

the inhibition rate is 1- (AUC of administration group/AUC of blank vehicle group) × 100%

(5) Statistical treatment

Experimental data were expressed using Mean ± standard error (Mean ± SEM), clinical scores were analyzed using One-way ANOVA, and significant differences were considered for p < 0.05.

2. Results of the experiment

Figure 1 and figure 2 show the AUC inhibition results for each group of clinical scores and clinical scores, and table 6 shows the AUC inhibition results for the clinical scores and clinical scores at the end of the experiment.

Table 6:

group of	Numbering	Clinical score (endpoint)	Clinical score AUC inhibition
				G1	Normal mice	0
G2	Blank solvent	8.5
				G3	Positive control group	4.4	42.3％
G4	002	2.3	80.3％

At the experimental endpoint (day 42), the clinical scores of the G3, G4 groups were significantly lower than the vehicle blank group. Clinical score AUC inhibition results from day 21 to day 42 show that compound 002 exhibited superior efficacy in inhibiting arthritis compared to the positive control group.

Although embodiments of the present disclosure have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present disclosure, and that changes, modifications, substitutions and alterations may be made to the above embodiments by those of ordinary skill in the art within the scope of the present disclosure.

Claims (12)

1. A compound which is a compound represented by the general formula (A-1) below or a stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof:

wherein the content of the first and second substances,

R¹selected from H, unsubstituted or optionally substituted by at least one R^aSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

the at least one R^aEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂Unsubstituted or optionally substituted by at least one R^bSubstituted of the following groups: -NH₂、C₁-C₁₀Alkyl radical、C₁-C₁₀Alkoxy radical, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, C₃-C₁₀Cycloalkyl radical, C₃-C₁₀Cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, C₆-C₂₀Aryl radical, C₆-C₂₀Aryloxy, 5-20 membered heteroaryl, 5-20 membered heteroaryloxy;

the at least one R^bEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂Or the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

R²selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂，

Provided that when R is²When is H, R¹Not methyl, ethyl, isopropyl.

2. A compound represented by the general formula (A), a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof:

wherein R is¹Selected from H, unsubstituted or optionally substituted by one or more R^aSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

each R^aIdentical or different from each other, independently of one another, selected from F, Cl, Br, I, OH, CN, ═ O, NO₂Unsubstituted or optionally substituted by one or more R^bSubstituted of the following groups: -NH₂、C₁-C₁₀Alkyl radical, C₁-C₁₀Alkoxy radical, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkenyloxy radical, C₂-C₁₀Alkynyl, C₂-C₁₀Alkynyloxy, C₃-C₁₀Cycloalkyl radical, C₃-C₁₀Cycloalkyloxy, 3-10 membered heterocyclyl, 3-10 membered heterocyclyloxy, C₆-C₂₀Aryl radical, C₆-C₂₀Aryloxy, 5-20 membered heteroaryl, 5-20 membered heteroaryloxy;

each R^bThe same or different, each is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂Or the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl, C₂-C₁₀Alkenyl radical, C₂-C₁₀Alkynyl, C₆-C₂₀Aryl, 5-20 membered heteroaryl;

wherein R is¹Not methyl, ethyl, isopropyl.

3. A compound according to claim 1 or 2, wherein in formula (A-1) or (A), R is¹Selected from unsubstituted or optionally substituted by at least one R^aSubstituted C₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl;

the at least one R^aEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂Or the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl.

4. The compound of claim 1, characterized in thatIn the general formula (A-1), R²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀Cycloalkyl, 3-10 membered heterocyclyl;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, CN, ═ O, NO₂、NH₂。

5. The compound of claim 1, wherein in the general formula (A-1), R is²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₁₀Alkyl radical, C₃-C₁₀A cycloalkyl group;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, ═ O.

6. The compound of claim 1, wherein in the general formula (A-1), R is²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₅Alkyl radical, C₃-C₅A cycloalkyl group;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, OH, ═ O.

7. The compound of claim 1, wherein in the general formula (A-1), R is²Selected from H, unsubstituted or optionally substituted by at least one R^cSubstituted of the following groups: c₁-C₅Alkyl radical, C₃-C₅A cycloalkyl group;

the at least one R^cEach of which is the same or different and is independently selected from F, Cl, Br, I, respectively.

8. The compound of claim 1 or 2, wherein the compound is selected from one of the following structures:

9. a pharmaceutical composition comprising a compound according to any one of claims 1 to 8.

10. Use of a compound according to any one of claims 1 to 8, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof, for the manufacture of a medicament for the treatment of a disease associated with PDE 4.

11. The use according to claim 10, wherein the PDE 4-associated disease is selected from the group consisting of inflammatory diseases, allergic diseases, autoimmune diseases, transplant rejection, joint inflammatory diseases, skin inflammatory diseases, inflammatory bowel diseases, and diseases associated with smooth muscle contractility.

12. Use according to claim 10 or claim 11, wherein the disease associated with PDE4 is selected from asthma, chronic bronchitis, chronic obstructive pulmonary disease, allergic rhinitis, adult respiratory distress syndrome, idiopathic dermatitis, psoriasis, urticaria, rheumatoid arthritis, osteoarthritis, gouty arthritis or spondylitis, ulcerative colitis, crohn's disease, overactive bladder.

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