CN113735825A - 1,2,3, 6-tetrahydropyridine compound and preparation method and application thereof - Google Patents

Abstract

The present invention provides a 1,2,3, 6-tetrahydropyridine compound, a pharmaceutical composition and use thereof, wherein the compound of the present invention not only has a good inhibitory effect on ADAMTS-5 and/or ADAMTS-4, but also has a weak inhibitory effect on MMP-2, and exhibits improved metabolic stability.

Description

1,2,3, 6-tetrahydropyridine compound and preparation method and application thereof

Technical Field

The present invention relates to 1,2,3, 6-tetrahydropyridines and their use for the treatment of ADAMTS-5 and/or ADAMTS-4 mediated diseases, such as osteoarthritis.

Background

Articular cartilage is a non-vascular tissue, 95% of its volume being occupied by an extracellular matrix, which is composed mainly of two components: type II collagen and aggrecan. Type II collagen is an extracellular matrix protein, forms a cross-linked rigid triple-helical structure, and ensures the mechanical strength of cartilage. Aggrecan is a extensively glycosylated extracellular matrix protein with three globular domains G1-G3 distributed from N-terminus to N-terminus with over 100 hydrophilic glycosyl side chains between the globular domains G2 and G3, so that saturated hydrated glycosyl groups ensure the toughness and elasticity of cartilage. The extracellular matrix is secreted by chondrocytes and degraded by a series of proteases. Under normal physiological conditions, extracellular matrix production and degradation are in dynamic equilibrium, maintaining normal cartilage homeostasis. When homeostasis is disrupted, the rate of extracellular matrix degradation exceeds the rate of synthesis, and cartilage exhibits degenerative structural and functional impairment, common diseases such as rheumatoid arthritis and osteoarthritis.

Among them, osteoarthritis is a disease characterized by degenerative changes of articular cartilage, accompanied by pain, joint distortion and dysfunction. According to analysis of a Global Data database, in 2018, the number of knee arthritis sufferers in seven main western developed countries is nearly 1.2 hundred million; the 2018 edition of osteoarthritis diagnosis and treatment guidelines newly released by the bone science society of the Chinese medical society shows that the prevalence rate of the domestic knee arthritis is 8.1% (about 1.1 hundred million people), and particularly, the incidence rate of the old aged over 65 years is over 50%. Therefore, osteoarthritis is a common disease worldwide, particularly occurs to the middle-aged and the elderly, seriously affects the quality of life of people, and brings heavy medical and economic burden to various countries.

Currently, the clinical use of drugs for osteoarthritis treatment is limited, and the main therapeutic approach is to use non-steroidal anti-inflammatory drugs (NSAIDS) to relieve pain and inflammation symptoms, which cannot block the disease mechanism, and when NSAIDS is no longer effective, the patient needs to undergo joint replacement surgery. Therefore, there is a great clinical need for an effective disease-modifying anti-osteoarthritis drug.

ADAMTS-5 (disintegrin metalloprotease-5 with thrombospondin motif-4, also known as aggrecanase-2) is a zinc ion metalloprotease that cleaves the peptide bond Glu373-Ala374 between globular domains of G1-G2 to release aggrecan fragments (ARGS), which are believed to be the first step in aggrecan degradation. As early as 2005, both the Stanton (Nature 2005,434(7033): 648-. However, by 2007, Song et al (Arthritis Rheum 2007,56(2):575-585) found that siRNA transfection interfering with the expression of either ADAMTS-5 or ADAMTS-4 in human chondrocytes reduced aggrecan degradation, but that simultaneous inhibition of ADAMTS-4 and ADAMTS-5 was comparable to the effect exhibited by ADAMTS-5 alone. On the other hand, ADAMTS-5 and ADAMTS-4 proteins have high homology and have specificity to degradation of aggrecan (J.Med.chem.2014,57(24):10476-10485), so that selective inhibition of ADAMTS-5 and/or ADAMTS-4 is expected to avoid skeletal muscle toxicity of other matrix metalloproteinase inhibitors, and the inhibition lays a foundation for development of ADAMTS-5 and/or ADAMTS-4 inhibitors for treating osteoarthritis.

Matrix Metalloproteinases (MMPs) constitute another 23 zinc metalloprotease family with many of the same structural elements as members of the ADAMTS family. Clinical studies in oncology with broad-spectrum MMP inhibitors have shown that inhibition of specific MMPs is associated with a poor prognosis and undesirable side effects. As early as 2002, Itoh et al (J Immunol 2002,169(5):2643-2647.) found that deletion of MMP-2 caused aggravation of arthritis by gene knockout experiments. Thus, inhibition of MMP-2 is undesirable in the development of ADAMTS-5 small molecule inhibitors.

In recent years, a few studies of ADAMTS-5 small molecule inhibitors have been reported, such as WO2014066151, WO2016102347, WO 2017211662. Similarly, the public data show that the only small molecule compound GLPG1972 in the current clinical stage has better in-vitro inhibition effect on ADAMTS-5, but the phase 2 clinical dosage of the compound reaches 150-600 mg per day, which indicates that the pharmacokinetic property of the compound has room for improvement.

Disclosure of Invention

The present invention provides compounds that are ADAMTS-5 and/or ADAMTS-4 inhibitors, that have good ADAMTS-5 and/or ADAMTS-4 inhibitory activity, that inhibit MMP-2 poorly, and that unexpectedly exhibit improved metabolic stability.

One aspect of the present invention provides a compound having the structure of formula I or formula II or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically-labeled compound, metabolite or prodrug thereof, wherein:

R¹independently selected from hydrogen, cyano, C_1-4Alkyl radical, C_2-4Alkenyl radical, C_2-4Alkynyl, C_3-7Monocyclic cycloalkyl, 4-7 membered monocyclic heteroalkyl having 1-2 heteroatoms independently selected from N, O, S, phenyl, 5-6 membered monocyclic heteroaryl having 1-2 heteroatoms independently selected from N, O, S; wherein said alkyl, alkenyl, alkynyl, monocyclic cycloalkyl, monocyclic heteroalkyl, phenyl, 5-6 membered monocyclic heteroaryl has 0-2 independently selected R⁸A substituent group;

x is-CR^7aR^7b-；

R^2a、R^2b、R³、R^4a、R^4bEach independently selected from hydrogen, halogen, C_1-3Alkyl radical, C_2-3Alkenyl or C_2-3An alkynyl group; or, R^2aAnd R^2bTogether with the carbon atom to which they are attached form a 3-5 membered cycloalkyl group; r^4aAnd R^4bTogether with the carbon atom to which they are attached form a 3-5 membered cycloalkyl group; said alkyl, alkenyl, alkynyl or cycloalkyl group being optionally substituted with 0-3 independent halogens;

R^5a、R^5beach independently selected from hydrogen, halogen, C_1-3Alkyl radical, C_2-3Alkenyl radical, C_2-3Alkynyl, C_1-3Alkoxy, allyloxy or propargyloxy; or, R^5aAnd R^5bTogether with the carbon atom to which they are attached form a 3-5 membered cycloalkyl or heterocycloalkyl group; the alkyl, alkenyl and alkynylAlkoxy, allyloxy, propargyloxy, cycloalkyl, heterocycloalkyl can be substituted with 0-3 independently optional halogens;

a is C_6-10Aryl or C_5-10A meta heteroaryl, said aryl and heteroaryl optionally substituted with n independently selected R⁶Substituted by groups;

n is 0, 1,2 or 3;

R⁶independently selected from halogen, cyano, C_1-3Alkyl radical, C_2-3Alkenyl radical, C_2-3Alkynyl, C_3-6Cycloalkyl radical, C_6-10Aryl radical, C_6-12Aralkyl, 5-14 membered heteroaryl, 4-10 membered heterocyclyl, -OR^9a、-SR^9a、-N(R^9a)(R^9b)、-N(R^9a)C(＝O)R^9b、-C(＝O)R^9a、-C(＝O)N(R^9a)(R^9b)、-N(R^9a)S(＝O)₂R^9b、-S(＝O)₂R^9a、-S(＝O)₂N(R^9a)(R^9b) Wherein said alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, and heterocyclyl are each optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, amino, cyano, oxo, C_1-6Alkyl radical, C_1-6Haloalkyl, C_3-6Cycloalkyl, 3-10 membered heterocyclyl, -O-C_1-6Alkyl, -O-C_1-6A haloalkyl group;

R^7aindependently selected from hydrogen, C_1-3An alkyl group optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, C_1-3An alkoxy group;

R^7bindependently selected from hydrogen, hydroxy, C_1-3Alkyl radical, C_1-3Alkoxy, each of said alkyl, alkoxy being optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, amino, C_1-3An alkoxy group;

R⁸independently selected from halogen, cyano, C_1-3Alkyl radical, C_2-3Alkenyl radical, C_2-3Alkynyl, C_3-6Cycloalkyl radical, C_6-10Aryl radical, C_6-12Aralkyl, 5-14 membered heteroaryl,3-10 membered heterocyclyl, -OR^9a、-SR^9a、-N(R^9a)(R^9b)、-N(R^9a)C(＝O)R^9b、-C(＝O)R^9a、-C(＝O)N(R^9a)(R^9b)、-N(R^9a)S(＝O)₂R^9b、-S(＝O)₂R^9a、-S(＝O)₂N(R^9a)(R^9b)；

R^9aAnd R^9bIndependently selected from hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-6Cycloalkyl radical, C_6-10Aryl radical, C_6-12Aralkyl, 5-14 membered heteroaryl, 3-10 membered heterocyclyl; wherein said alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, and heterocyclyl are each optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, amino, cyano, oxo, C_1-6Alkyl radical, C_1-6Haloalkyl, C_3-6Cycloalkyl, 3-10 membered heterocyclyl, -O-C_1-6Alkyl, -O-C_1-6A haloalkyl group.

In a preferred embodiment, a compound of formula I or formula II according to the invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically-labelled compound, metabolite or prodrug thereof, R thereof¹Independently selected from hydrogen, C_1-4Alkyl radical, C_3-7Monocyclic cycloalkyl, phenyl, 5-6 membered monocyclic heteroaryl having 1-2 heteroatoms independently selected from N, O, S; wherein, the alkyl, monocyclic cycloalkyl, phenyl and monocyclic heteroaryl are optionally substituted by halogen or C_1-3Alkyl substitution.

In another preferred embodiment, a compound of formula I or formula II according to the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, wherein R is¹Independently selected from hydrogen, C_1-4Alkyl radical, C_3-7Monocyclic cycloalkyl, phenyl, 5-6 membered monocyclic heteroaryl having 1-2 heteroatoms independently selected from N, O, S; wherein said alkyl, monocyclic cycloalkyl, phenyl, monocyclic heteroaryl are optionally substituted with halo、C_1-3Alkyl substitution.

In another preferred embodiment, a compound of formula I or formula II according to the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically-labelled compound, metabolite or prodrug thereof, R¹Cyclopropyl is preferred.

In another preferred embodiment, a compound of formula I or formula II according to the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, wherein R is^7a、R^7bIndependently selected from hydrogen.

In another preferred embodiment, a compound of formula I or formula II according to the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, wherein R is^2a、R^2b、R³、R^4a、R^4bEach independently selected from hydrogen.

In another preferred embodiment, a compound of formula I or formula II, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, according to the invention is a phenyl or a 5-10 membered monocyclic or fused bicyclic heteroaryl group comprising 1,2 or 3 heteroatoms independently selected from N, O and S;

said aryl and heteroaryl groups being optionally substituted by n independently selected R⁶Substituted by groups;

n is 0, 1,2 or 3;

R⁶independently selected from halogen, cyano, C_1-3An alkyl group.

In another preferred embodiment, the compound of formula I or formula II according to the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, wherein a is phenyl, pyridine, pyrimidine, thiophene, pyrazole, imidazole, quinoline;

the phenyl, pyridine, pyrimidine, thiophene, pyrazole and imidazoleOxazole, quinoline optionally substituted with n independently selected R₆Substituted by groups;

n is 0, 1,2 or 3;

R₆independently selected from halogen, cyano, C_1-3An alkyl group.

In another preferred embodiment, a compound of formula I or formula II according to the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically-labelled compound, metabolite or prodrug thereof, wherein a is phenyl; the phenyl is optionally substituted with 0, 1,2 or 3 halogens.

In another preferred embodiment, the compound of formula I or formula II according to the present invention, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, wherein the compound is selected from the group consisting of:

another aspect of the invention provides a pharmaceutical composition comprising a prophylactically or therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers.

Another aspect of the present invention provides the use of a compound of the present invention, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention, in the manufacture of a medicament for the prevention or treatment of an ADAMTS-5 and/or ADAMTS-4 mediated disease or disorder.

Another aspect of the present invention provides a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention, for use in a method for preventing or treating an ADAMTS-5 and/or ADAMTS-4 mediated disease or disorder, the method comprising administering to a subject in need thereof an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention.

In a preferred embodiment, the ADAMTS-5 and/or ADAMTS-4 mediated disease or disorder is selected from the group consisting of osteoarthritis, rheumatoid arthritis, psoriatic arthritis, gonococcal arthritis, high fever arthritis, yersinia arthritis, gouty arthritis, pyrophosphate arthritis, suppurative arthritis, and articular cartilage damage and degenerative changes resulting from clinical glucocorticoid overuse, among others.

Definition of

Unless defined otherwise below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. Reference to the techniques used herein is intended to refer to those techniques commonly understood in the art, including those variations of or alternatives to those techniques that would be apparent to those skilled in the art. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

As used herein, the terms "comprises," "comprising," "has," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

As used herein, the term "hydrogen" and hydrogen in each group refers to protium (H), deuterium (D), or tritium (T).

As used herein, the term "alkyl" is defined as a straight or branched chain monovalent saturated aliphatic hydrocarbon group. C_1-12Alkyl means an alkyl group having 1 to 12 carbon atoms, for example 1 to 6 carbon atoms (C)_1-6Alkyl) or 1 to 4 carbon atoms (C)_1-4Alkyl groups). For example, as used herein, the term "C_1-6Alkyl "refers to a linear or branched radical of 1 to 6 carbon atomsA group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or n-hexyl) which is optionally substituted with 1 or more (such as 1 to 3) suitable substituents such as halo (in which case the group is referred to as "haloalkyl") (e.g., CH)₂F、CHF₂、CF₃、CCl₃、C₂F₅、C₂Cl₅、CH₂CF₃、CH₂Cl or-CH₂CH₂CF₃Etc.). The term "C1-4 alkyl" refers to a linear or branched aliphatic hydrocarbon chain of 1 to 4 carbon atoms (i.e., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl).

The term "alkenyl" as used herein means a straight or branched chain monovalent hydrocarbon radical containing one or more double bonds and having from 2 to 6 carbon atoms ("C)_2-6Alkenyl "). The alkenyl group includes, but is not limited to, ethenyl, 1-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, and 4-methyl-3-pentenyl. When the compounds of the invention contain alkenyl groups, the compounds may be present in the pure E (entgegen) configuration, in the pure Z (ipsilateral (zusammen)) configuration or as a mixture thereof in any proportion.

As used herein, the term "alkynyl" refers to a monovalent hydrocarbon group containing one or more triple bonds, which may be straight or branched, including but not limited to ethynyl, 1-propynyl, 3-propynyl, and the like.

As used herein, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic ring, including but not limited to spiro, fused, or bridged systems (such as bicyclo [ 1.1.1)]Pentyl, bicyclo [2.2.1]Heptyl, bicyclo [3.2.1]Octyl or bicyclo [5.2.0]Nonyl, decalinyl, etc.), optionally substituted with 1 or more (such as 1 to 3) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term“C_3-6Cycloalkyl "refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring of 3 to 6 ring-forming carbon atoms (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) optionally substituted with 1 or more (such as 1 to 3) suitable substituents, for example, methyl-substituted cyclopropyl.

As used herein, the term "heterocyclyl" refers to a saturated or partially unsaturated monovalent monocyclic or bicyclic group having 2,3, 4,5, 6, 7, 8 or 9 carbon atoms in the ring and one or more (e.g., one, two, three or four) selected from C (═ O), O, S, S (═ O), S (═ O)₂And NR^aWherein R is^aRepresents a hydrogen atom or C_1-6Alkyl or C_1-6A haloalkyl group; the heterocyclic group may be attached to the rest of the molecule through any of the carbon atoms or nitrogen atom (if present). In particular, a 3-to 10-membered heterocyclyl group is a group having 3-10 carbon atoms and heteroatoms in the ring, including, but not limited to, oxiranyl, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, dioxolyl, pyrrolidinyl, pyrrolidinonyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl.

As used herein, the term "aryl" refers to an all-carbon monocyclic or fused ring polycyclic aromatic group having a conjugated pi-electron system. For example, as used herein, the term "C_6-14Aryl "means an aromatic group containing 6 to 14 carbon atoms, such as phenyl or naphthyl. Aryl is optionally substituted with 1 or more (such as 1 to 3) suitable substituents (e.g. halogen, -OH, -CN, -NO)₂、C_1-6Alkyl, etc.).

As used herein, the term "aralkyl" denotes an aryl-substituted alkyl group, wherein the aryl group and the alkyl group are as defined herein. Typically, the aryl group can have 6 to 14 carbon atoms and the alkyl group can have 1 to 6 carbon atoms. Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, phenylbutyl.

As used herein, the term "5-14 membered heteroaryl" refers to a monocyclic aromatic group containing 5-14 ring members, and the ring members contain at least 1 (e.g., 1,2,3, or 4) heteroatoms selected from N, O and S, e.g., "5-6 membered heteroaryl", 5-membered heteroaryl, 6-membered heteroaryl, and the like. Specific examples thereof include, but are not limited to, furyl, thienyl, pyrrolyl, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, imidazolyl, pyrazolyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,2, 5-oxadiazolyl, 1, 3, 4-oxadiazolyl, pyridyl, 2-pyridonyl, 4-pyridonyl, pyrimidinyl, 2H-1, 2-oxazinyl, 4H-1, 2-oxazinyl, 6H-1, 2-oxazinyl, 4H-1, 3-oxazinyl, 6H-1, 3-oxazinyl, 4H-1, 4-oxazinyl, pyridazinyl, pyrazinyl, 1,2, 3-triazinyl, 1, 3, 5-triazinyl, 1,2, 4, 5-tetrazinyl, and the like.

As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

As used herein, the term "substituted" means that one or more (e.g., one, two, three, or four) hydrogens on the designated atom is replaced with a selection from the indicated group, provided that the designated atom's normal valency at the present time is not exceeded and the substitution results in a stable compound. Combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

If a substituent is described as "optionally substituted", the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of the list of substituents, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be optionally replaced individually and/or together with an independently selected substituent. If the nitrogen of a substituent is described as being optionally substituted with one or more of the list of substituents, then one or more hydrogens on the nitrogen (to the extent any hydrogen present) may each be optionally replaced with an independently selected substituent.

If a substituent is described as being "independently selected from" a group, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

As used herein, the term "one or more" means 1 or more than 1, such as 2,3, 4,5 or 10, under reasonable conditions.

As used herein, unless otherwise indicated, the point of attachment of a substituent may be from any suitable position of the substituent.

When a bond of a substituent is shown through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

The invention also includes all pharmaceutically acceptable isotopically-labeled compounds, which are identical to those of the present invention, except that one or more atoms are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number prevailing in nature. Examples of isotopes suitable for inclusion in compounds of the invention include, but are not limited to, isotopes of hydrogen (e.g., deuterium (g), (b), (c), (d) and (d)²H) Tritium (a)³H) ); isotopes of carbon (e.g. of¹¹C、¹³C and¹⁴C) (ii) a Isotopes of chlorine (e.g. of chlorine)³⁶Cl); isotopes of fluorine (e.g. of fluorine)¹⁸F) (ii) a Isotopes of iodine (e.g. of iodine)¹²³I and¹²⁵I) (ii) a Isotopes of nitrogen (e.g. of¹³N and¹⁵n); isotopes of oxygen (e.g. of¹⁵O、¹⁷O and¹⁸o); isotopes of phosphorus (e.g. of phosphorus)³²P); and isotopes of sulfur (e.g. of³⁵S). Certain isotopically-labeled compounds of the present invention (e.g., those into which a radioisotope is incorporated) are useful in drug and/or substrate tissue distribution studies (e.g., assays). Radioisotope tritium (i.e. tritium³H) And carbon-14 (i.e.¹⁴C) Are particularly useful for this purpose because of their ease of incorporation and ease of detection. Using positron-emitting isotopes (e.g. of the type¹¹C、¹⁸F、^{1 5}O and¹³n) can be used in Positron Emission Tomography (PET) studies for testing substrate substratesVolume occupancy. Isotopically labeled compounds of the present invention can be prepared by processes analogous to those described in the accompanying schemes and/or in the examples and preparations by using an appropriate isotopically labeled reagent in place of the non-labeled reagent employed previously. Pharmaceutically acceptable solvates of the invention include those in which the crystallization solvent may be isotopically substituted, e.g., D₂O, acetone-d₆Or DMSO-d₆。

The term "stereoisomer" denotes an isomer formed as a result of at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, they can give rise to racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers. Certain individual molecules may also exist as geometric isomers (cis/trans). Similarly, the compounds of the invention may exist as mixtures of two or more structurally different forms (commonly referred to as tautomers) in rapid equilibrium. Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It is understood that the scope of this application encompasses all such isomers or mixtures thereof in any ratio (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

Solid (—), solid wedge shapes may be used herein

Or virtual wedge shape

Chemical bonds of the compounds of the present invention are depicted. The use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers (e.g., particular enantiomers, racemic mixtures, etc.) at that carbon atom are included. The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the stereoisomers shown are present. When present in the racemic mixture, the acid addition salt,solid and imaginary wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, the compounds of the present invention are intended to exist as stereoisomers, including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotamers, conformers, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerization and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

The present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

It will also be appreciated that certain compounds of the invention may be present in free form for use in therapy or, where appropriate, in the form of a pharmaceutically acceptable derivative thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, solvates, N-oxides, metabolites or prodrugs, which upon administration to a patient in need thereof are capable of providing, directly or indirectly, a compound of the present invention or a metabolite or residue thereof. Thus, when reference is made herein to "a compound of the invention," it is also intended to encompass the various derivative forms of the compounds described above.

Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.

Suitable acid addition salts are formed from acids which form pharmaceutically acceptable salts. Examples include aspartate, glucoheptonate, gluconate, orotate, palmitate and other similar salts.

Suitable base addition salts are formed from bases which form pharmaceutically acceptable salts. Examples include aluminum salts, arginine salts, choline salts, magnesium salts, and other similar salts.

For a review of suitable Salts see Stahl and Wermuth, "Handbook of Pharmaceutical Salts: properties, Selection, and Use "(Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

The compounds of the invention may be present in the form of solvates, preferably hydrates, wherein the compounds of the invention comprise as structural element of the crystal lattice of the compound a polar solvent, such as in particular water, methanol or ethanol. The amount of polar solvent, particularly water, may be present in stoichiometric or non-stoichiometric proportions.

Those skilled in the art will appreciate that not all nitrogen-containing heterocycles are capable of forming N-oxides, since the available lone pair is required for oxidation of the nitrogen to the oxide; one skilled in the art will recognize nitrogen-containing heterocycles that are capable of forming N-oxides. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include oxidation of heterocycles and tertiary amines with peroxy acids such as peracetic and m-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkyl hydroperoxides such as t-butyl hydroperoxide, sodium perborate, and dioxiranes (dioxiranes) such as dimethyldioxirane. These methods for preparing N-oxides have been widely described and reviewed in the literature, see for example: T.L.Gilchrist, Comprehensive Organic Synthesis, vol.7, pp 748-750; a.r.katitzky and a.j.boulton, eds., Academic Press; and G.W.H.Cheeseman and E.S.G.Werstuk, Ad lance in Heterocyclic Chemistry, vol.22, pp 390-.

Also included within the scope of the present invention are metabolites of the compounds of the present invention, i.e., substances formed in vivo upon administration of the compounds of the present invention. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc. of the administered compound. Accordingly, the present invention includes metabolites of the compounds of the present invention, including compounds made by the process of contacting the compounds of the present invention with a mammal for a time sufficient to produce a metabolite thereof.

The present invention further includes within its scope prodrugs of the compounds of the present invention which are certain derivatives of the compounds of the present invention which may themselves have little or no pharmacological activity which, when administered into or onto the body, may be converted to the compounds of the present invention having the desired activity by, for example, hydrolytic cleavage. Typically such prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the desired therapeutically active compound. Additional information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems", volume 14, ACS Symposium Series (t.higuchi and v.stella). Prodrugs of the invention may be prepared, for example, by substituting certain moieties known to those skilled in the art as "pro-moieties" (e.g., "Design of Prodrugs", described in h. bundgaard (Elsevier, 1985)) for appropriate functional groups present in compounds of the invention.

The invention also encompasses compounds of the invention containing a protecting group. In any process for preparing the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved by conventional protecting Groups, such as those described in T.W.Greene & P.G.M.Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. The protecting group may be removed at a suitable subsequent stage using methods known in the art.

The term "about" means within. + -. 10%, preferably within. + -. 5%, more preferably within. + -. 2% of the stated value.

By "pharmaceutically acceptable carrier" in the context of the present invention is meant a diluent, adjuvant, excipient, or vehicle that is administered together with a therapeutic agent and which is, within the scope of sound medical judgment, suitable for contact with the tissues of humans and/or other animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit/risk ratio.

Pharmaceutically acceptable carriers that may be employed in the pharmaceutical compositions of the present invention include, but are not limited to, sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is an exemplary carrier when the pharmaceutical composition is administered intravenously. Physiological saline and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like. The composition may also optionally contain minor amounts of wetting agents, emulsifying agents, or pH buffering agents. Oral formulations may contain standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate and the like. Examples of suitable pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (1990).

The pharmaceutical compositions of the present invention may act systemically and/or locally. For this purpose, they may be administered by a suitable route, for example by injection (e.g. intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular injection, including instillation) or transdermally; or by oral, buccal, nasal, transmucosal, topical, in the form of ophthalmic preparations or by inhalation.

For these routes of administration, the pharmaceutical compositions of the present invention may be administered in suitable dosage forms.

Such dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups.

The term "effective amount" as used herein refers to an amount of a compound that, when administered, will alleviate one or more symptoms of the condition being treated to some extent.

The dosing regimen may be adjusted to provide the best desired response. For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is noted that dosage values may vary with the type and severity of the condition being alleviated, and may include single or multiple doses. It is further understood that for any particular individual, the specific dosage regimen will be adjusted over time according to the individual need and the professional judgment of the person administering the composition or supervising the administration of the composition.

Detailed Description

In order to make the objects and technical solutions of the present invention clearer, the present invention is further illustrated below with reference to specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, specific experimental methods not mentioned in the following examples were carried out according to the usual experimental methods.

The abbreviations herein have the following meanings:

meaning of abbreviations

TLC thin layer chromatography

LC-MS liquid chromatography-mass spectrometry combination

DMF N, N-dimethylformamide

DMSO dimethyl sulfoxide

EA Ethyl acetate

PE Petroleum Ether

THF tetrahydrofuran

LDA lithium diisopropylamide

EDCI 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

HOBT 1-hydroxybenzotriazole

Pd(dppf) Cl₂[1, 1' -bis (diphenylphosphino) ferrocene]Palladium dichloride

SFC supercritical fluid chromatography

The structures of the compounds described in the following examples were measured by nuclear magnetic resonance spectroscopy (¹H-NMR) or Mass Spectrometry (MS).

¹The H-NMR analyzer is Bruker 400MHz NMR spectrometer, and the solvent is deuterated methanol (CD)₃OD), deuterated chloroform (CDCl)₃) Or hexadeuterated dimethyl sulfoxide (DMSO-d6) with the internal standard Tetramethylsilane (TMS). Chemical shifts (δ) are given in parts per million ppm).

The Mass Spectrometer (MS) was an Agilent (ESI) mass spectrometer model Agilent 6120B.

Thin Layer Chromatography (TLC) was carried out using an aluminum plate (20X 20cm) from Merck, and thin layer preparative chromatography was carried out using a GF254(0.4 to 0.5mm) silica gel plate.

The reaction was monitored by Thin Layer Chromatography (TLC) or liquid chromatography-mass spectrometry (LC-MS) using a developing reagent system including dichloromethane and methanol systems, n-hexane and ethyl acetate systems, and petroleum ether and ethyl acetate systems. The developer system is adjusted according to the polarity of the compound to be separated (by adjusting the volume ratio of the solvent or adding triethylamine or the like).

The instrument model used for preparing the high performance liquid chromatography is as follows: agilent 1260, column: waters Xbridge Prep C18OBD (19 mm. times.150 mm. times.5.0 μm); temperature of the chromatographic column: 25 ℃; flow rate: 20.0 mL/min; detection wavelength: 214 nm; elution gradient: (0 min: 10% A, 90% B; 16.0 min: 90% A, 10% B); mobile phase A: 100% acetonitrile; mobile phase B: 0.05% aqueous ammonium bicarbonate solution.

Unless otherwise stated, the reaction temperature is room temperature (20 ℃ C. to 30 ℃ C.).

The reagents used in the examples were purchased from Acros Organics, Aldrich Chemical Company, Shanghai Teber Chemical science and technology, Inc., and the like.

Reference example intermediate preparation

Synthesis of intermediate (S) -3- (4-cyclopropyl-2, 5-dioxoimidazolin-4-yl) propionic acid (Int1)

Step 1: preparation of tert-butyl 4-cyclopropyl-4-oxobutyrate (Int1-2)

Under the protection of nitrogen, a solution of cyclopropylmethyl ketone Int1-1(30g,356.63mmol) in THF (30mL) was added dropwise to a-78 ℃ solution of LDA (45.84g,427.96mmol,214mL) in THF (150mL), and after completion of the addition, the reaction was warmed to 20 ℃ and stirred for 30 minutes. Then, the reaction solution was cooled again to-78 ℃ and a solution of tert-butyl 2-bromoacetate (69.56g,356.63mmol, Bide) in THF (30mL) was slowly added thereto, stirred at ordinary temperature,overnight. After completion of the reaction, the reaction was quenched with a saturated aqueous ammonium chloride solution (150mL), the reaction solution (100 mL. times.3) was extracted with EA, and the organic phase was washed with a saturated saline solution and Na₂SO₄After drying, filtration and evaporation of the filtrate under reduced pressure gave Int1-2 as a crude yellow oil (65.4g,329.88mmol, 92.50% yield).¹H NMR(400MHz,CDCl₃):δ2.83(t,J＝6.8Hz,2H),2.50(t,J＝6.8Hz,2H),1.97-1.92(m,1H),1.45(s,9H),1.06-1.01(m,2H),0.91-0.86(m,2H)。

Step 2: preparation of tert-butyl 3- (4-cyclopropyl-2, 5-dioxoimidazolin-4-yl) propionate (Int1-3)

The intermediate Int1-2(19.8g,100mmol), ammonium carbonate (76.8g,800mmol), sodium cyanide (12.25g,250mmol), ethanol (150mL) and water (150mL) were sequentially added to the reactor, stirred and heated to 80 ℃ for reaction for 18 hours, the reaction solution was cooled to room temperature, the reaction solution was extracted with 300mL of ethyl acetate and 300mL of water, the aqueous phase was extracted with ethyl acetate several times (200mL × 3), the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate and filtered, and the concentrated organic phase was separated with SGC (EA/PE ═ 1/2) to obtain the target intermediate Int1-3(17.1g,63.72mmol, 51.36% yield) as a white solid.¹H NMR(400MHz,DMSO)δ10.61(s,1H),7.66(s,1H),2.29-2.08(m,2H),1.93-1.88(m,2H),1.29(s,9H),1.09-1.02(m,1H),0.47-0.26(m,3H),0.11-0.04(m,1H)。

And step 3: preparation of 3- (4-cyclopropyl-2, 5-dioxoimidazolin-4-yl) propionic acid (Int1-4)

Intermediate Int1-3(19.8g,100mmol) was added to a 1, 4-dioxane solution of hydrogen chloride (4M,150mL), stirred at room temperature for 4 hours and concentrated, and the precipitated solid was triturated in acetonitrile (100mL) for 1 hour and filtered to give purified racemate Int1-4 as a white solid.

And 4, step 4: preparation of (S) -3- (4-cyclopropyl-2, 5-dioxoimidazolin-4-yl) propionic acid (Int1)

Intermediate Int1-4 was isolated by SFC to give Int1(6.3g,29.72mmol, 46.63% yield, chiral purity 99.16% ee) as a white solid.¹H NMR(400MHz,DMSO):δ12.20(s,1H),10.63(s,1H),7.71(s,1H),2.32-2.09(m,2H),1.99-1.87(m,2H),1.11-1.03(m,1H),0.48-0.27(m,3H),0.12-0.05(m,1H)。ESI-MS m/z 213.1[M+1]⁺。

Separation conditions are as follows:

type of chiral column: CHIRALPAK AD-H10 um 2.5 x 25 cm;

flow rate: 70 g/min;

mobile phase supercritical carbon dioxide: methanol 60: 40;

detection wavelength: 214 nm;

retention time: 2.915 minutes.

Synthesis of intermediate 5-aminomethyl-5-cyclopropylimidazoline-2, 4-dione hydrochloride (Int2)

Step 1: preparation of tert-butyl (2-cyclopropyl-2-oxoethyl) carbamate (Int2-2)

A THF solution of cyclopropylmagnesium bromide (26.99g,192.44mmol) was added dropwise to a 10 ℃ solution of Int2-1(14g,64.15mmol) in THF (30mL), and after dropwise addition, the mixture was stirred at 10 ℃ for 2 hours. After completion of the reaction monitored by TLC (PE/EA ═ 3/1), the reaction was quenched with 1N hydrochloric acid solution, however, the reaction solution was extracted several times with ethyl acetate, and the combined organic phases were washed with saturated brine and anhydrous Na₂SO₄After drying, filtration and concentration of the filtrate, the intermediate Int2-2 was isolated as a yellow oil by silica gel column chromatography (EA/PE ═ 1/10) (8g,40.15mmol, 62.59% yield).

Step 2: preparation of t-butyl ((4-cyclopropyl-2, 5-dioxoimidazolin-4-yl) methyl) carbamate (Int2-3)

Adding the intermediate Int2-2(10.8g,54.20mmol), ammonium carbonate (26.02g,271.02mmol), potassium cyanide (7.50g,108.41mmol), methanol (100mL) and water (50mL) into a reactor in sequence, stirring and heating to 80 ℃ for reaction for 16 hours, cooling the reaction liquid to room temperature, extracting the reaction liquid with 200mL of ethyl acetate and 350mL of water, extracting the aqueous phase with ethyl acetate for multiple times, washing the combined organic phase with saturated brine, drying and filtering with anhydrous sodium sulfate, grinding the concentrated organic phase in a mixed solution of 15mL of petroleum ether and 10mL of ethyl acetate for 1 hour, and filtering to obtain the target intermediate Int2-3(5.7g,21.17mmol, 39.05% yield) as a white solid.

And step 3: preparation of 5-aminomethyl-5-cyclopropylimidazoline-2, 4-dione hydrochloride (Int2)

Intermediate Int2-3(210mg,0.854mmol) was added to a solution of hydrogen chloride in 1, 4-dioxane (5mL), and after stirring at room temperature for 1 hour, the precipitated solid was filtered to give purified Int2(140mg,0.68mmol, 87.5% yield) as a white solid.¹H NMR(400MHz,DMSO-d6)δ10.78(s,1H),8.26(brs,3H),7.82(s,1H),3.13-3.09(m,1H),2.93-2.89(m,1H),1.04-0.97(m,1H),0.46-0.32(m,2H),0.29-0.23(m,1H),0.03-0.00(m,1H).ESI-MS m/z 170.3[M+H]⁺。

Synthesis of intermediate 4- (3, 5-difluorophenyl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3a)

Step 1: preparation of tert-butyl 4- (3, 5-difluorophenyl) -3, 6-dihydro-2H-pyridine-1-carboxylate (Int3a-1)

3, 5-difluoro-1-bromobenzene (500mg,2.59mmol), N-Boc-1,2,3, 6-tetrahydropyridine-4-boronic acid pinacol ester (800m g,2.59mmol), cesium carbonate (2.52g,7.77mmol) and Pd (dppf) Cl₂(189mg,0.259mmol) was charged into a reaction flask, dioxane and water (40mL/8mL) were added, and the mixture was stirred under nitrogen and warmed to 100 ℃ for 12 hours. Cooling to room temperature, adding saturated ammonium chloride aqueous solution (50mL) and ethyl acetate (40mL) for extraction, extracting the aqueous phase with ethyl acetate for multiple times, washing the combined organic phases with saturated saline (50mL), drying with anhydrous sodium sulfate, filtering, evaporating the filtrate under reduced pressure to obtain a crude product, and purifying the crude product by silica gel column chromatography (eluent: petroleum ether/ethyl acetate 5:1v/v) to obtain 4- (3, 5-difluorophenyl) -3, 6-dihydropyridine-2H-1-carboxylic acid tert-butyl ester (Int3a-1) as a light yellow solid, wherein the yield is 72.4%, and ESI-MS M/z 240.2[ M-55-difluorophenyl) -3, 6-dihydropyridine-2H-1-carboxylic acid (M-a-1)]⁺。

Step 2: preparation of 4- (3, 5-difluorophenyl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3a)

Int3a-1(250mg,0.847mmol) was added to the reaction flask and dissolved in 2mL of ethyl acetateThen, an ethyl acetate hydrochloride solution (7mL, about 2.5M) was added thereto, and the reaction was stirred at room temperature for 4 hours. The precipitated solid was collected by suction filtration, and the filter cake was washed with ethyl acetate (2mL) and dried in vacuo to give 4- (3, 5-difluorophenyl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3a) as a white solid in 72.8% yield, ESI-MS M/z 196.3[ M +1 ]]⁺。

Synthesis of intermediate 4- (3, 5-dichloro-phenyl) -1,2,3, 6-tetrahydro-pyridine hydrochloride (Int3b)

Step 1: synthesis of 4- (3, 5-dichlorophenyl) -N-Boc-1,2,3, 6-tetrahydropyridine (Int3 b-1):

to a 100mL single-neck flask were added 3, 5-dichlorobromobenzene (0.501g, 2.2mmol), N-Boc-1,2,3, 6-tetrahydropyridine-4-boronic acid pinacol ester (0.688g, 2.2mmol), Pd (dppf) Cl₂(0.164g, 0.22mmol), cesium carbonate (1.501g, 4.6mmol), 1, 4-dioxane 10mL, and 1mL of pure water were replaced with nitrogen gas three times, the mixture was stirred and heated to 100 ℃ to keep the temperature for reaction for 6 hours, after the completion of the TLC monitoring reaction, the mixture was cooled to room temperature, 50mL of pure water was added to the reaction solution, and extracted with ethyl acetate (50mL × 3) several times, the organic phases were combined, washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, evaporated to dryness under reduced pressure, and finally purified by silica gel column chromatography (100 mesh to 200 mesh silica gel, PE eluent: EA ═ 30: 1 to 10: 1) to obtain 4- (3,5 dichlorophenyl) -N-Boc-1,2,3, 6-tetrahydropyridine as a pale yellow oily substance (0.67g, yield: 92.25%).¹H NMR(400MHz,CDCl₃)δ7.24(dd,J＝4.2,1.5Hz,3H),6.08(s,1H),4.08(s,2H),3.62(t,J＝5.6Hz,2H),2.46(s,2H),1.49(s,9H).

Step 2: synthesis of 4- (3, 5-dichlorophenyl) -1,2,3, 6-tetrahydropyridine (Int3 b):

4- (3, 5-dichlorophenyl) -N-Boc-1,2,3, 6-tetrahydropyridine (0.291g, 0.89mmol) and 1.5mL of ethyl acetate were sequentially added to a 50mL single-neck flask, and stirred at room temperature for 5 minutes, then 5mL of a hydrogen chloride-ethyl acetate solution (2.5mol/L) was added, and the reaction was stirred at room temperature for 3 hours; after TLC detection reaction is completed, filtering is carried out, a filter cake is washed by ethyl acetate, and a solid is dried under reduced pressureDrying gave 0.21g of 4- (3, 5-dichlorophenyl) -1,2,3, 6-tetrahydropyridine as a white solid (yield: 89.13%). ESI-MS M/z 228.4(M + H)⁺。

Synthesis of intermediate 4- (3, 4-dichloro-phenyl) -1,2,3, 6-tetrahydro-pyridine hydrochloride (Int3c)

Step 1: synthesis of 4- (3, 4-dichlorophenyl) -N-Boc-1,2,3, 6-tetrahydropyridine (Int3c-1)

The target compound Int3c-1 was prepared in a similar manner to the above synthesis of intermediate Int3b, step 1 (pale yellow oil, yield 73.98%).¹H NMR(400MHz,DMSO-d6)δ7.68(d,J＝2.1Hz,0H),7.60(d,J＝8.5Hz,0H),7.44(dd,J＝8.5,2.2Hz,0H),6.29(s,0H),4.00(s,0H),3.52(t,J＝5.7Hz,2H),2.45(s,0H),1.42(s,9H)。

Step 2: synthesis of 4- (3, 4-dichlorophenyl) -1,2,3, 6-tetrahydropyridine:

the target compound Int3c (10.18g, white solid yield 82.67%) was prepared in a similar manner to the above synthesis of intermediate Int3b, step 2. ESI-MS M/z 228.2(M + H)⁺。

Synthesis of intermediate 4- (5-fluoro-3-chlorophenyl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3d)

Step 1: preparation of Synthesis of 4- (5-fluoro-3-chlorophenyl) -N-Boc-1,2,3, 6-tetrahydropyridine (Int3d-1)

Using a method analogous to step 1 of the synthesis of intermediate Int3b, the title compound Int3d-1(265mg, colorless oil, 87.84% yield) was prepared.

Step 2: preparation of 4- (3-chloro-5-fluorophenyl) -1,2,3, 6-tetrahydro-pyridine hydrochloride (Inte3d)

The target compound Int3d was prepared for use in a similar manner as described above for Int3b intermediate synthesis step 2.

Preparation of intermediate 4- (1-methyl-1H-pyrazol-3-yl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3e)

Step 1: preparation of 4- (1-methyl-1H-pyrazol-3-yl) -3, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (Int3e-1)

Using a similar procedure as described above for the synthesis of intermediate Int3b, step 1, 260mg of the target compound Int3e-1 was prepared as a colorless oil in 61.02% yield.

Step 2: preparation of 4- (1-methyl-1H-pyrazol-3-yl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3e)

The target compound Int3e was prepared for use in a similar manner as described above for Int3b intermediate synthesis step 2.

Preparation of intermediate 4- (1-methyl-1H-pyrazol-3-yl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3f)

Step 1: preparation of 5-methyl-1-isopropyl-3-bromo-1H-pyrazole (Int3f-1)

Sodium cyanide (0.745g, 0.018mol) was placed in a reaction flask, 60mL of DMF was added and stirred and the temperature was reduced to 0 ℃. 5-methyl-3-bromo-1H-pyrazole (22.078g, 0.012mol) in DMF (8mL) was slowly added to the reaction mixture, the temperature was raised to 50 ℃ to react for 30 minutes, the temperature was again lowered to 0 ℃ and a solution of isopropyl iodide (2.60g, 0.015mol) in DMF (10mL) was slowly added to the reaction mixture to react at room temperature for 15 hours. After the reaction is monitored to be completed, 80mL of saturated ammonium chloride solution and 80mL of ethyl acetate are added into the reaction solution for extraction, the aqueous phase is extracted by 40mL of ethyl acetate again, the organic phase obtained by combination is dried and filtered by anhydrous sodium sulfate, the crude product is obtained by reduced pressure evaporation to dryness, and the crude product is purified by silica gel column chromatography (petroleum ether/ethyl acetate is 5:1) to obtain the pure product, the yield is 16.4%, and ESI-MS M/z 203.0[ M +1 ]]⁺。

Step 2: preparation of 4- (5-methyl-1-isopropyl-1H-pyrazol-3-yl) -3, 6-dihydropyridine-2H-1-carboxylic acid tert-butyl ester (Int3f-2)

The target compound Int3f-2 was prepared in a similar manner as described above for Int3b intermediate synthesis step 1, as a pale yellow solid with 69.8% yield. ESI-MS M/z 306.2[ M +1 ]]⁺。

And step 3: preparation of 4- (1-isopropyl-5-methyl-1H-pyrazol-3-yl) -1,2,3, 6-tetrahydro-pyridine hydrochloride (Int3f)

The target compound Int3f was prepared in a similar manner as the above intermediate Int3b, step 2, as a pale yellow solid in 69.8% yield. ESI-MS M/z 206.0[ M +1 ]]⁺。

Preparation of intermediate 4- (1, 5-dimethyl-1H-pyrazol-3-yl) -1,2,3, 6-tetrahydropyridine hydrochloride (Int3g)

Step 1: preparation of 4- (1, 5-dimethyl-1H-pyrazol-3-yl) -3, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester (Int3g-1)

The target compound Int3g-1 is prepared by a method similar to the synthesis step 1 of the intermediate Int3b, and the character and the yield are 63%. ESI-MS M/z 222.2[ M-55 ]]⁺。

Step 2: preparation of 4- (1, 5-dimethyl-1H-pyrazol-3-yl) -1,2,3, 6-tetrahydropyridine hydrochloride

The target compound Int3g is prepared by adopting a method similar to the synthesis step 2 of the intermediate Int3b, and the character and the yield are 89%. ESI-MS M/z 178.1[ M +1 ]]⁺。

Preparation of intermediate 5- (1,2,3, 6-tetrahydropyridin-4-yl) -thiophene-2-carbonitrile trifluoroacetic acid salt (Int3h)

Step 1: preparation of 4- (5-cyano-thiophen-2-yl) -3, 6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester (Int3H-1)

The synthesis step of the intermediate Int3b is adopted1 analogously, the target compound Int3h-1 was obtained in 38.6% yield. ESI-MS M/z 235.2[ M-55 ]]⁺。

Step 2: preparation of 5- (1,2,3, 6-tetrahydropyridin-4-yl) -thiophene-2-carbonitrile trifluoroacetic acid salt (Int3h)

The crude target compound Int3h was prepared in 98.7% yield by a method similar to the above synthesis step 2 of intermediate Int3 b. ESI-MS M/z 191.1[ M +1 ]]⁺。

Preparation of intermediate 5- (1,2,3, 6-tetrahydropyridin-4-yl) -thiophene-2-carbonitrile trifluoroacetic acid salt (Int3i)

Step 1: preparation of 4- (quinolin-3-yl) -3, 6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester (Inti-1)

The target compound Int3i-1 is prepared by a method similar to the synthesis step 1 of the intermediate Int3b, and the character and the yield are 74%. ESI-MS M/z 235.2[ M-55 ]]⁺。

Step 2: preparation of 3- (1,2,3, 6-tetrahydropyridin-4-yl) quinoline (Int3i)

The target compound Int3i is prepared by a method similar to the synthesis step 2 of the intermediate Int3b, and the character and yield are 98.7%. ESI-MS M/z 191.1[ M +1 ]]⁺。

Preparation of intermediate 5- (1,2,3, 6-tetrahydropyridin-4-yl) -thiophen-2-methyltrifluoroacetate (Int3j)

Step 1: preparation of 4- (5-methylthiophen-2-yl) -3, 6-dihydro-2H-pyridine-1-carboxylic acid tert-butyl ester (Int3j-1)

The target compound Int3j-1 was prepared in 30.2% yield using a similar procedure as described above for the synthesis of intermediate Int3b, step 1. ESI-MS M/z 224.1[ M-55 ]]⁺。

Step 2: preparation of 5- (1,2,3, 6-tetrahydropyridin-4-yl) -thiophen-2-methyltrifluoroacetate (Int3j)

The crude target compound Int3j was prepared in 97% yield using a method similar to the above synthesis of intermediate Int3b, step 2. ESI-MS M/z 180.1[ M +1 ]]⁺。

Example 1: preparation of (S) -5-cyclopropyl-5- {3- [4- (3, 5-difluoro-phenyl) -3, 6-dihydro-2H-pyridin-1-yl ] -3-oxo-propyl } -imidazoline-2, 4-dione

3- (4-cyclopropyl-2, 5-dioxo-imidazolin-4-yl) -propionic acid (50mg, 0.24mmol), 4- (3, 5-difluoro-phenyl) -1,2,3, 6-tetrahydro-pyridine hydrochloride (57.2mg, 0.247mmol), EDCI (67.9mg, 0.353mmol), HOBT (48.1mg, 0.353mmol) and triethylamine (71.5mg, 0.710mmol) were dissolved in 10mL of dichloromethane and the reaction was stirred at room temperature for 12 hours. To the reaction was added 20mL of dichloromethane and 30mL of saturated ammonium chloride solution, the aqueous phase was extracted twice more with dichloromethane (20mL), the organic phases were combined, washed with saturated sodium chloride (30mL × 2), dried over anhydrous sodium sulfate, filtered and dried, and the resulting crude product was separated on a thin layer chromatography plate (developing solvent: dichloromethane/methanol ═ 12:1v/v), purified and lyophilized to give the product 5-cyclopropyl-5- {3- [4- (3, 5-difluoro-phenyl) -3, 6-dihydro-2H-pyridin-1-yl ] -3-oxo-propyl } -imidazoline-2, 4-dione as a white solid in 34.9% yield.

¹H NMR(400MHz,DMSO-d6)δ10.63(d,J＝5.6Hz,1H),7.74(d,J＝11.2Hz,1H),7.24–7.17(m,2H),7.17–7.10(m,1H),6.37(d,J＝4.0Hz,1H),4.18-4.08(m,2H),3.70–3.63(m,1H),3.60(t,J＝5.6Hz,1H),2.49-2.38(m,2H),2.37–2.22(m,1H),2.03–1.91(m,2H),1.29-1.21(m,1H),1.16–1.04(m,1H),0.50-0.41(m,1H),0.39-0.28(m,2H),0.17–0.08(m,1H)。

ESI-MS m/z 390.2[M+H]⁺。

Using a synthetic procedure similar to that described above in example 1 and known to those skilled in the art, the compounds shown in table 1 were prepared:

TABLE 1

Biological evaluation

Experimental example 11: hADAMTS-5 enzyme activity inhibition assay

1.1 Experimental materials

Human ADAMTS-5(hADAMTS-5) was purchased from R & D systems (cat # 2198-AD-020), and WAAG-3R was purchased from Anaspec (cat # AS-60431-1);

72% polyvinyl lauroyl oxide (Brij-35) was purchased from Shanghai Biotechnology engineering Co., Ltd. (product No. A600638), p-Aminomercuric acetate (APMA) was purchased from Sigma (product No. A9563), Tris was purchased from AMRESCO (product No. 0497- & 500G), and the other reagents were of AR grade.

384 microplates purchased from grignard (GBO, cat # 781901);

the type of the microplate reader: SpectraMax M5 multifunctional microplate reader.

1.2 Experimental methods

The detection method is improved according to the product specification, and specifically comprises the following steps: preparing an analysis buffer solution: 50mM Tris,100mM NaCl,5mM CaCl₂0.05% Brij-35, pH 7.5. mu.L of compound diluent (final concentration 2mM, 1/4 diluted in assay buffer), 10. mu.L of hADAMTS-5 enzyme reaction mixture (final concentration 0.5. mu.g,assay buffer dilution), incubation for 30min at room temperature. After 30min, 5. mu.L of the reaction substrate WAAG-3R (final concentration 25. mu.M) was added to each well for color development. Measuring fluorescence value of each well at excitation wavelength of 340nm and emission wavelength of 420nm in dynamic mode of microplate reader, comparing with blank well without sample to be measured, and calculating inhibition rate of compound on enzyme [ inhibition rate ═ 1-sample group fluorescence value/blank group fluorescence value) × 100%]IC50 values were calculated in a Prism GraphPad using a four parameter model. Duplicate wells were set for each assay and each set of experiments was repeated twice.

1.3 results of the experiment

The inhibitory effect of the compounds on hADAMTS-5 activity was determined as described above, and some results are shown in Table 2

TABLE 2

Examples	IC50(nM)
		1	14.7
2	33.7
		8	136.9
9	25.7

1.4 conclusion of the experiment

The compounds of each example have strong inhibition effect on hADAMTS-5.

Experimental example 12: hMMP-2 enzyme activity inhibition assay

1.1 Experimental materials

Humanized MMP-2(hMMP-2) from R&D systems (cat # 902-MP-010), substrate Mca-PLGL-Dpa-AR-NH₂From R&D systems (cat # ES 001).

1.2 Experimental methods

The detection method is improved according to the product specification, and specifically comprises the following steps: preparing an analysis buffer solution: 50mM Tris,10mM CaCl₂150mM NaCl, 0.05% Brij-35, pH 7.5. Preparing hMMP-2 enzyme pre-activation solution: a final concentration of 10ng (assay buffer dilution) was pre-activated by incubation with freshly prepared 1mM APMA for 1h at 37 ℃. mu.L of compound dilutions (final concentration of 5mM, 1/4 diluted in assay buffer) and 10. mu.L of hMMP-2 enzyme pre-activator were added sequentially to 384 plates and incubated at room temperature for 30 min. After 30min, 5. mu.L of the reaction substrate MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (final concentration 10. mu.M) was added to each well for color development. Measuring fluorescence value of each well at 320nm excitation wavelength and 405nm emission wavelength in kinetic mode of microplate reader, comparing with blank well without sample to be measured, and calculating inhibition rate of compound on enzyme [ inhibition rate ═ 1-sample group fluorescence value/blank group fluorescence value) × 100%]IC50 values were calculated in a Prism GraphPad using a four parameter model. Duplicate wells were set for each assay and each set of experiments was repeated twice.

1.3 results of the experiment

The inhibitory effect of the compounds on MMP-2 activity was determined as described above, and some results are shown in Table 3

TABLE 3

Examples	IC50(nM)
		2	1375.4
9	421.8

1.4 conclusion of the experiment

The compounds of each example had a weak effect on hMMP-2 inhibition.

Experimental example 13: study of hepatic microsome stability in SD rat

1.1 Experimental methods

The test compound (2. mu.L) was mixed with rat liver microsome solution (100. mu.L), and after preincubation at 37 ℃ for 10 minutes, NADPH (. mu.L) was added and incubated for 0, 5, 10, 20, 30, and 60 minutes at incubation concentrations of 1. mu.M, 1 unit/mL, and 0.5mg/mL for the test compound, NADPH, and liver microsome enzymes. After termination of the reaction by addition of glacial acetonitrile (600. mu.L) an appropriate volume of internal standard was added, vortexed, centrifuged (4000 rpm, 4 ℃, 20 min) and the supernatant (300. mu.L) was aspirated. And (3) measuring the residual concentration of the sample in the supernatant at different incubation times, and plotting Ln (drug residual amount%) "to the incubation time to obtain a rate constant, thereby calculating the half-life period and the liver clearance of the drug, and evaluating the metabolic stability of the drug in the liver microsome by using the half-life period and the liver clearance value of the drug.

1.2 results of the experiment

TABLE 4 results of hepatic microsome stability experiments

Is a compound represented by number 255 in patent WO2016102347

1.3 conclusion of the experiment:

the experimental data in the table show that the intrinsic clearance rate and half-life period of rat liver of the compound 2 are obviously reduced and the stability is obviously improved compared with the compound R255.

Experimental example 14: pharmacokinetic study of SD rat

1.1 Experimental methods

The compounds of the invention were administered to male SD rats by Intravenous (IV) and intragastric (PO) administration, respectively, and pharmacokinetic profiles were examined. IV and PO were administered at 1mg/kg and 5mg/kg (or 10mg/kg), respectively, in 5% DMSO: 20% Solutol: 75% physiological saline. Blood was collected at various time points after IV and PO administration, anticoagulated with EDTAK2, centrifuged to obtain plasma samples, stored at-80 ℃. Plasma samples were processed for precipitated protein and analyzed by LC-MS/MS. Pharmacokinetic parameters were calculated using Phoenix WinNonlin 6.3 software using a non-compartmental model, and the results are shown in Table 5.

1.2 results of the experiment

TABLE 5 results of pharmacokinetic experiments in vivo in rats

Is a compound representative of patent number 255 in WO2016102347, prepared according to the patented process; AUC is the area under the curve when drug is applied, and represents the exposure of the drug in rats; cmax is the maximum blood concentration; f is bioavailability.

1.3 conclusion of the experiment

As shown in table 5, compound 2 of the present invention has good in vivo exposure upon administration either IV or PO, and in particular, has an oral bioavailability as high as 66.9%, significantly higher than the bioavailability of the representative compound No. 255 in patent WO2016102347, which is 10.2%.

While specific embodiments of the invention have been described in detail, it will be appreciated by those skilled in the art that various modifications and alternatives to those details could be developed in light of the overall teachings of the disclosure, and that such modifications are intended to be within the scope of the disclosure. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims (10)

1. A compound of the structure of formula I or formula II or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically-labeled compound, metabolite, prodrug thereof, wherein:

wherein,

R¹independently selected from hydrogen, cyano, C_1-4Alkyl radical, C_2-4Alkenyl radical, C_2-4Alkynyl, C_3-7Monocyclic cycloalkyl, 4-7 membered monocyclic heteroalkyl having 1-2 heteroatoms independently selected from N, O, S, phenyl, 5-6 membered monocyclic heteroaryl having 1-2 heteroatoms independently selected from N, O, S; wherein said alkyl, alkenyl, alkynyl, monocyclic cycloalkyl, monocyclic heteroalkyl, phenyl, 5-6 membered monocyclic heteroaryl has 0-2 independently selected R⁸A substituent group;

x is-CR^7aR^7b-；

R^2a、R^2b、R³、R^4a、R^4bEach independently selected from hydrogen, halogen, C_1-3Alkyl radical, C_2-3Alkenyl or C_2-3An alkynyl group; or, R^2aAnd R^2bTogether with the carbon atom to which they are attached form a 3-5 membered cycloalkyl group; r^4aAnd R^4bTogether with the carbon atom to which they are attached form a 3-5 membered cycloalkyl group; said alkyl, alkenyl, alkynyl or cycloalkyl group being optionally substituted with 0-3 independent halogens;

R^5a、R^5beach independently selected from hydrogen, halogen, C_1-3Alkyl radical, C_2-3Alkenyl radical, C_2-3Alkynyl, C_1-3Alkoxy, allyloxy or propargyloxy; or, R^5aAnd R^5bTogether with the carbon atom to which they are attached form a 3-5 membered cycloalkyl or heterocycloalkyl group; the alkyl, alkenyl, alkynyl, alkoxy, allyloxy, propargyloxy, cycloalkyl, heterocycloalkyl can be substituted with 0-3 independent optional halogens;

a is C_6-10Aryl or 5-10 membered heteroaryl, said aryl and heteroaryl optionally substituted with n independently selected R⁶Substituted by groups; n is 0, 1,2 or 3;

R⁶independently selected from halogen, cyano, C_1-3Alkyl radical, C_2-3Alkenyl radical, C_2-3Alkynyl, alkynyl,C_3-6Cycloalkyl radical, C_6-10Aryl radical, C_6-12Aralkyl, 5-14 membered heteroaryl, 4-10 membered heterocyclyl, -OR^9a、-SR^9a、-N(R^9a)(R^9b)、-N(R^9a)C(＝O)R^9b、-C(＝O)R^9a、-C(＝O)N(R^9a)(R^9b)、-N(R^9a)S(＝O)₂R^9b、-S(＝O)₂R^9a、-S(＝O)₂N(R^9a)(R^9b) Wherein said alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, and heterocyclyl are each optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, amino, cyano, oxo, C_1-6Alkyl radical, C_1-6Haloalkyl, C_3-6Cycloalkyl, 3-10 membered heterocyclyl, -O-C_1-6Alkyl, -O-C_1-6A haloalkyl group;

R^7aindependently selected from hydrogen, C_1-3An alkyl group optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, C_1-3An alkoxy group;

R^7bindependently selected from hydrogen, hydroxy, C_1-3Alkyl radical, C_1-3Alkoxy, each of said alkyl, alkoxy being optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, amino, C_1-3An alkoxy group;

R⁸independently selected from halogen, cyano, C_1-3Alkyl radical, C_2-3Alkenyl radical, C_2-3Alkynyl, C_3-6Cycloalkyl radical, C_6-10Aryl radical, C_6-12Aralkyl, 5-14 membered heteroaryl, 3-10 membered heterocyclyl, -OR^9a、-SR^9a、-N(R^9a)(R^9b)、-N(R^9a)C(＝O)R^9b、-C(＝O)R^9a、-C(＝O)N(R^9a)(R^9b)、-N(R^9a)S(＝O)₂R^9b、-S(＝O)₂R^9a、-S(＝O)₂N(R^9a)(R^9b)；

R^9aAnd R^9bIndependently selected from hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, alkynyl,C_3-6Cycloalkyl radical, C_6-10Aryl radical, C_6-12Aralkyl, 5-14 membered heteroaryl, 3-10 membered heterocyclyl; wherein said alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heteroaryl, and heterocyclyl are each optionally substituted with 0-3 substituents independently selected from the group consisting of: halogen, hydroxy, amino, cyano, oxo, C_1-6Alkyl radical, C_1-6Haloalkyl, C_3-6Cycloalkyl, 3-10 membered heterocyclyl, -O-C_1-6Alkyl, -O-C_1-6A haloalkyl group.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, prodrug thereof, wherein:

R¹independently selected from hydrogen, C_1-4Alkyl radical, C_3-7Monocyclic cycloalkyl, phenyl, 5-6 membered monocyclic heteroaryl having 1-2 heteroatoms independently selected from N, O, S; wherein, the alkyl, monocyclic cycloalkyl, phenyl and monocyclic heteroaryl are optionally substituted by halogen or C_1-3Alkyl substitution.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, prodrug thereof, wherein:

R¹independently selected from hydrogen, C_1-4Alkyl radical, C_3-7Monocyclic cycloalkyl, phenyl, 5-6 membered monocyclic heteroaryl having 1-2 heteroatoms independently selected from N, O, S; wherein, the alkyl, monocyclic cycloalkyl, phenyl and monocyclic heteroaryl are optionally substituted by halogen or C_1-3Alkyl substitution; preferably, R¹Selected from cyclopropyl.

4. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, prodrug thereof, wherein:

R^7a、R^7bindependently selected from hydrogen.

5. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, prodrug thereof, wherein: r^2a、R^2b、R³、R^4a、R^4bEach independently selected from hydrogen.

6. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, prodrug thereof, wherein:

a is phenyl or a 5-10 membered monocyclic or fused bicyclic heteroaryl group containing 1,2 or 3 heteroatoms independently selected from N, O and S, said aryl and heteroaryl groups optionally being substituted with n independently selected R⁶Substituted by groups;

n is 0, 1,2 or 3;

R⁶independently selected from halogen, cyano, C_1-3An alkyl group.

7. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, prodrug thereof, wherein:

a is phenyl, pyridine, pyrimidine, thiophene, pyrazole, imidazole, quinoline, optionally substituted with n independently selected R₆Substituted by groups; n is 0, 1,2 or 3; r₆Independently selected from halogen, cyano, C_1-3An alkyl group;

preferably, a is phenyl; the phenyl is optionally substituted with 0, 1,2 or 3 halogens.

8. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically-labeled compound, metabolite, prodrug thereof, wherein compound is selected from the group consisting of:

9. a pharmaceutical composition comprising a compound of any one of claims 1-8, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically-labeled compound, metabolite, prodrug thereof, and one or more pharmaceutically acceptable excipients.

10. Use of a compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, prodrug or pharmaceutical composition thereof, for the manufacture of a medicament for the prevention or treatment of an ADAMTS-5 and/or ADAMTS-4 mediated disease or disorder, preferably, an ADAMTS-5 and/or ADAMTS-4 mediated disease or disorder selected from the group consisting of osteoarthritis, rheumatoid arthritis, psoriatic arthritis, gonococcal arthritis, hyperthermic arthritis, yersinia arthritis, gouty arthritis, pyrophosphate arthritis, suppurative arthritis, and articular cartilage damage or degenerative lesion caused by clinical glucocorticoid overuse.