HK1241267A1

HK1241267A1 - Compounds and methods for inducing chondrogenesis

Info

Publication number: HK1241267A1
Application number: HK18100555.7A
Authority: HK
Inventors: 彼得‧G‧舒尔茨; 阿纳布‧K‧查特吉; 守天‧朱; 乔舒亚‧帕耶特; 洪哲‧尹; 百元‧杨
Original assignee: 斯克利普斯研究所
Priority date: 2013-03-15
Filing date: 2018-01-15
Publication date: 2018-06-08

Abstract

Described herein are compounds and compositions for the amelioration of arthritis or joint injuries by inducing mesenchymal stem cells into chondrocytes.

Description

Compounds and methods for inducing chondrogenesis

This application is a divisional application of chinese patent application No. 201480028421.3 (international application No. PCT/US2014/026722) entitled "compounds and methods for inducing cartilage formation" filed on 13/03 of 2014.

Cross Reference to Related Applications

This application claims the benefit of U.S. application No. 61/794,094 filed on 3, 15, 2014, which is incorporated herein by reference in its entirety.

Technical Field

The present invention relates to compounds, compositions, articles of manufacture and uses thereof for inducing cartilage formation and for treating arthritis or joint injury.

Background

Osteoarthritis (OA) is the most common disorder of muscle bone. Approximately 4 million americans currently suffer from the disease, and due to aging of the population and an extended life expectancy, this figure is expected to increase to 6 million over the next twenty years, making it the fourth leading cause of disability. OA is characterized by slow degenerative breakdown of the joint, which includes articular cartilage (containing cells and matrix that produce lubrication and buffer for the joint) and subchondral bone underlying the articular cartilage. Current OA therapies include pain relief by oral NSAIDs or selective cyclooxygenase 2(COX-2) inhibitors, intra-articular (IA) injection of agents such as corticosteroids and hyaluronic acid, and surgical procedures.

Mesenchymal Stem Cells (MSCs) are present in adult articular cartilage and can be programmed in vitro after isolation to differentiate into chondrocytes and other mesenchymal cell lineages. In part, it is regulated by growth factors (TGF, BMP), serum conditions, and cell-cell contacts.

Disclosure of Invention

Provided herein is a method of ameliorating arthritis or joint damage in a mammal comprising administering to a joint of the mammal a composition having a therapeutically effective amount of a compound disclosed herein.

Provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound disclosed herein to induce differentiation of stem cells into chondrocytes.

In one aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula I:

wherein:

each R¹Independently is halo, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted aryloxy, CN, NO₂、SR⁴、S(O)R⁴、SO₂R⁴、NR⁴R¹¹、CO₂H or CO₂R⁴；

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴、CH₂CO₂H、CH₂CO₂R⁴Or optionally substituted phenyl;

y is a bond, - (CR)⁵R⁶)-、-(CR⁷R⁸)(CR⁹R¹⁰) -or- (CR)⁷R⁸)(CR⁹R¹⁰)X-；

X is O or CR⁵R⁶；

R²Is halo, C (O) R⁴、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴Or C (═ NOR)⁴)R⁴；

Each R³Independently selected from H, CN, halo, C (O) R⁴、CO₂H、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴；

Each R⁴Independently selected from H and optionally substituted alkyl;

each R⁵、R⁶、R⁷、R⁸、R⁹And R¹⁰Independently selected from H, halo, optionally substituted alkyl, OH, CO₂R⁴、NR⁴R¹¹And optionally substituted alkoxy; and is

R¹¹Is H, optionally substituted alkyl, C (O) R⁴、C(O)OR⁴、C(O)NR⁴R⁴Or SO₂R⁴；

Provided that

a) If Y is a bond and m is 0, then R²Selected from C (O) NR⁴R¹¹、(CR⁷R⁸)OR⁴、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴(ii) a And is

R²Not C (O) NH₂、p-CH₂OR⁴、p-CH(OH)CH₂OH、p-CH₂CH₂OH or p-CH₂CH₂CH₂OH; and is

b) The compound is not selected from

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula Ia:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

Each R³Independently selected from CN, halo, C (O) R⁴、CO₂H、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴；

X is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

each R⁵、R⁶、R⁷、R⁸、R⁹And R¹⁰Independently selected from H, halo, optionally substituted alkyl, OH, NR⁴R¹¹And optionally substituted alkoxy; and is

With the proviso that the compound is not selected from

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula Ib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

R²Is C (O) NR⁴R¹¹、(CR⁷R⁸)OR⁴、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴Or C (═ NOR)⁴)R⁴；

R³Is H;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

Provided that if n is 0, R²Not C (O) NH₂、p-CH₂OR⁴、p-CH(OH)CH₂OH、p-CH₂CH₂OH or p-CH₂CH₂CH₂OH。

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula Ic, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴；

Y is- (CR)⁵R⁶)-；

C is aryl or heteroaryl;

x is O or CR⁵R⁶；

R²Is halo, C (O) R⁴、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂R⁴、SO₂NH₂、SO₃H、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴Or C (═ NOR)⁴)R⁴；

Each R³Independently selected from H, CN, halo, C (O) R⁴、CO₂H、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

In another aspect, provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula I:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴、CH₂CO₂H、CH₂CO₂R³Or optionally substituted phenyl;

X is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

Provided that

b) The compound is not selected from

In another aspect, provided herein is a method of inducing differentiation of a mesenchymal stem cell into a chondrocyte, the method comprising contacting a mesenchymal stem cell with a compound of formula Ia:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

X is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

In another aspect, provided herein is a method of inducing differentiation of a mesenchymal stem cell into a chondrocyte, the method comprising contacting a mesenchymal stem cell with a sufficient amount of a compound of formula Ib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

R³Is H;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that if n is 4 and R¹Is H, then R²Not C (O) NH₂、p-CH₂OR⁴、p-CH(OH)CH₂OH、p-CH₂CH₂OH or p-CH₂CH₂CH₂OH。

In another aspect, provided herein is a method of inducing differentiation of a mesenchymal stem cell into a chondrocyte, the method comprising contacting a mesenchymal stem cell with a sufficient amount of a compound of formula Ic, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴；

Y is- (CR)⁵R⁶)-；

C is aryl or heteroaryl;

x is O or CR⁵R⁶；

Each R³Independently selected from H, CN, halo, C (O) R⁴、CO₂H、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

In some embodiments described above or below for compounds of formula I or Ia:

R²is halo, C (O) R⁴Alkyl, optionally substituted alkoxy, haloalkyl, (CR)⁷R⁸)OR⁴、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)C(O)OR⁴Or X (CR)⁷R⁸)C(O)NR⁴R¹¹(ii) a And is

Each R³Independently selected from CN, halo, C (O) R⁴、CO₂H、C(O)NR⁴R¹¹Alkyl or optionally substituted alkoxy;

or R³To adjacent R³Or with R²Together forming a ring.

In certain embodiments of the compounds of formula I or Ia described above or below:

R²is F, Cl, C (O) CH₃、CH₃、CF₃、OCH₃、OEt、OPr、OCF₃、OCHF₂、(CR⁷R⁸)OR⁴、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)C(O)OR⁴Or X (CR)⁷R⁸)C(O)NR⁴R¹¹(ii) a And is

Each R³Independently selected from CN, F, Cl, C (O) CH₃、CO₂H、C(O)NH₂、CH₃、OCF₃Or OCH₃；

Or R³To adjacent R³Or with R²Together forming a ring.

In certain embodiments, R³Independently selected from CN, F, Cl, C (O) CH₃Or CO₂H. In certain embodiments, R²Is F, Cl, C (O) CH₃、CH₃、CF₃、OCH₃、OEt、OPr、OCF₃Or CH₂CH₂CH₂OH。

In some embodiments described above or below for compounds of formula Ib:

R²is (CR)⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴Or C (═ NOR)⁴)R⁴(ii) a And is

R³Is H.

In certain embodiments, R²Is (CR)⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹Or (CR)⁷R⁸)NR⁴SO₂R⁴. In certain embodiments, R²Is CH₂CH₂OH、CH₂CH₂OCH₃、CH₂CHCH₃OH、CHCH₃CH₂OH、CH₂CH₂CH₂OH、CH₂CH₂CH₂NH₂、CH₂CH₂CHCH₃OH、C(CH₃)₂CH₂CH₂OH、CH₂CH₂C(CH₃)₂OH、OCH₂CH₂OH、OCH₂CH₂OCH₃Or OCH₂CH₂NH₂. In certain embodiments, R²Is (CR)⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴Or X (CR)⁷R⁸)C(O)NR⁴R¹¹. In certain embodiments, R²Is CH₂C(O)CH₃、CH₂C(O)NH₂、CH₂CH₂C(O)CH₃Or CH₂CH₂C(O)NH₂。

In some embodiments described above or below for compounds of formula Ic, C is aryl. In certain embodiments, C is phenyl.

In some embodiments described above or below for compounds of formula Ic, C is heteroaryl. In certain embodiments, C is pyridinyl, pyrimidinyl, pyridazinyl, or pyrazinyl.

In some embodiments described above or below for compounds of formula Ic:

R²is halo, C (O) R⁴、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂NH₂、SO₃H、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)OR⁴Or X (CR)⁷R⁸)C(O)NR⁴R¹¹(ii) a And is

Each R³Independently selected from H, CN, halo, CO₂H or haloalkyl.

In certain embodiments described above or below for compounds of formula Ic:

R²is Cl, F, C (O) CH₃、CO₂H、C(O)NR⁴R¹¹、CH₃Optionally substituted alkoxy, CF₃、SO₂NH₂、SO₃H、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)OR⁴Or X (CR)⁷R⁸)C(O)NR⁴R¹¹(ii) a And is

Each R³Independently selected from H, CN, Cl, F, CO₂H or CF₃。

In certain embodiments, R²Is Cl, F, C (O) CH₃、CO₂H、CH₃、OCH₃、CF₃(ii) a And each R³Independently selected from H, CN or CO₂H. In certain embodiments, R²Is CH₂C(O)NH₂、CH₂C(O)CH₃、CH₂C(O)OH、CH₂CH₂C (O) OH or CH₂CH₂C(O)NH₂。

In one aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula II:

wherein

Each R¹Independently is halo, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted aryloxy, CN, NO₂、SR⁴、S(O)R⁴、SO₂R⁴、NHR⁵、NR⁴R⁵、CO₂H or CO₂R⁴；

n is 0, 1,2, 3 or 4;

b is NHC (O) R²、NR³C(O)R²、NHC(O)NH₂、NHC(O)NHR²、NHC(O)NR²R⁴、NR³C(O)NH₂、NR³C(O)NHR²、NR³C(O)NR²R⁴、NHC(O)OR²、NR³C(O)OR²、NHSO₂R³、NR³SO₂R³、NHSO₂R⁴、NR³SO₂R⁴、NHSO₂NH₂、NHSO₂NHR²、NHSO₂NR²R⁴、NR³SO₂NH₂、NR³SO₂NHR²Or NR³SO₂NR²R⁴；

Each R²And R⁴Independently is optionally substituted phenyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted aralkyl or optionally substituted alkyl;

R³is optionally substituted alkyl or optionally substituted alkylA substituted aralkyl group;

R⁵is H, optionally substituted alkyl, C (O) R⁴、C(O)OR⁴、C(O)NR⁴R⁴Or SO₂R⁴；

A is CO₂H、CO₂R³、C(O)NH₂、C(O)NHR²、C(O)NR²R⁴Or SO₂NR^aR^b(ii) a And is

Each R^aAnd R^bIndependently an optionally substituted alkyl group or together with the N to which they are attached form a ring;

provided that

a) If B is NHC (O) R²Or NR³C(O)R²Then A is not CO₂H; and is

b) The compound is not selected from

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula IIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

Each R¹Independently isHalo, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted aryloxy, CN, NO₂、SR⁴、S(O)R⁴、SO₂R⁴、NHR⁵、NR⁴R⁵、CO₂H or CO₂R⁴；

n is 0, 1,2, 3 or 4;

b is NHC (O) NH₂、NHC(O)NHR²、NHC(O)NR²R⁴、NR³C(O)NH₂、NR³C(O)NHR²Or NR³C(O)NR²R⁴；

R³is optionally substituted alkyl or optionally substituted aralkyl;

R⁵is H, optionally substituted alkyl, C (O) R⁴、C(O)OR⁴、C(O)NR⁴R⁴Or SO₂R⁴(ii) a And is

A is CO₂H or CO₂R³；

With the proviso that the compound is not

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula lib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

b is NHC (O) R²Or NR³C(O)R²；

R²Is optionally substituted phenyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted aralkyl or optionally substituted alkyl;

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is SO₂NR^aR^b(ii) a And is

Each R^aAnd R^bIndependently an optionally substituted alkyl group or together with the N to which they are attached form a ring.

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula IIc, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

b is NHSO₂R³、NR³SO₂R³、NHSO₂R⁴、NR³SO₂R⁴、NHSO₂NH₂、NHSO₂NHR²、NHSO₂NR²R⁴、NR³SO₂NH₂、NR³SO₂NHR²Or NR³SO₂NR²R⁴；

each R³Independently an optionally substituted alkyl group or an optionally substituted aralkyl group;

A is C (O) NHR²Or C (O) NR²R⁴；

With the proviso that the compound is not

In another aspect, provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula II:

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

provided that

a) If B is NHC (O) R²Or NR³C(O)R²Then A is not CO₂H; and is

b) The compound is not selected from

In another aspect, provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula IIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is CO₂H or CO₂R³；

With the proviso that the compound is not

In another aspect, provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula lib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

b is NHC (O) R²Or NR³C(O)R²；

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is SO₂NR^aR^b(ii) a And is

In another aspect, provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula IIc, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

A is C (O) NHR²Or C (O) NR²R⁴；

With the proviso that the compound is not

In some embodiments of the compounds of formula IIa described above or below, B is NHC (O) NHR²、NHC(O)NR²R⁴、NR³C(O)NHR²Or NR³C(O)NR²R⁴. In certain embodiments, B is NHC (O) NHR²Or NR³C(O)NHR². In certain embodiments, B is NHC (O) NR²R⁴Or NR³C(O)NR²R⁴. In some embodimentsIn the case, B is NHC (O) NHR²。

In some embodiments of the compounds of formula IIa, described above or below, R²Is optionally substituted phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, R²Is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

In some embodiments of the compounds of formula IIb described above or below, B is NHC (O) R²。

In some embodiments of the compounds of formula IIb described above or below, B is NR³C(O)R². In certain embodiments, R³Is an optionally substituted alkyl group.

In some embodiments of the compounds of formula IIb described above or below, each R^aAnd R^bIndependently an optionally substituted alkyl group. In some embodiments of the compounds of formula IIb described above or below, R^aAnd R^bForm a ring together with the N to which they are attached.

In some embodiments of the compounds of formula IIb described above or below, R²Is optionally substituted phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, R²The substitution on the phenyl group of (A) is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

In some embodiments of the compounds of formula IIc described above or below, B is NHSO₂R³、NR³SO₂R³、NHSO₂R⁴Or NR³SO₂R⁴. In certain embodiments, B is NHSO₂R³Or NR³SO₂R³. In certain embodiments, B is NHSO₂R³. In certain embodiments, R³Is an optionally substituted alkyl group. In certain embodiments, R³Is CH₃. In certain embodiments, B is NHSO₂R⁴Or NR³SO₂R⁴. In certain embodiments, R⁴Is optionally substituted phenyl.

In some embodiments of the compounds of formula IIc described above or below, B is NHSO₂NH₂、NHSO₂NHR²、NHSO₂NR²R⁴、NR³SO₂NH₂、NR³SO₂NHR²Or NR³SO₂NR²R⁴。

In some embodiments of the compounds of formula IIc described above or below, A is C (O) NHR². In some embodiments of the compounds of formula IIc described above or below, A is C (O) NR²R⁴. In certain embodiments, R²Is optionally substituted phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, R²The substitution on the phenyl group of (A) is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula III, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

x is O, NH or NR⁶；

A is C (O), CH₂Or CH-CR³R⁴-C(O)R²；

R²Is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl;

R⁶Is optionally substituted phenyl;

provided that

a) If A is CH-CR³R⁴-C(O)R²X is O or NH;

b) if n is 0, A is CHCH₂C(O)R²And X is O, then R²Is not provided withAnd is

c) If A is C (O) or CH₂X is NR⁶And R is⁶Is not provided with

In another aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula IIIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

R²is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl; and is

R⁵Is H, optionally substituted alkyl, C (O) R⁴、C(O)OR⁴、C(O)NR⁴R⁴Or SO₂R⁴。

In another aspect, provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula III:

wherein

n is 0, 1,2, 3 or 4;

x is O, NH or NR⁶；

A is C (O), CH₂Or CH-CR³R⁴-C(O)R²；

R²Is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl;

R⁶Is optionally substituted phenyl;

provided that

a) If A is CH-CR³R⁴-C(O)R²X is O or NH;

b) if n is 0, A is CHCH₂C(O)R²And X is O, then R²Is not provided withAnd is

c) If A is C (O) or CH2, X is NR⁶And R is⁶Is not provided with

In another aspect, provided herein is a method of inducing differentiation of a mesenchymal stem cell into a chondrocyte, the method comprising contacting a mesenchymal stem cell with a sufficient amount of a compound of formula IIIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

R²is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl; and is

In some embodiments of the compounds of formula III described above or below, X is NR⁶And A is C (O). In some embodiments of the compounds of formula III described above or below, X is NR⁶And A is CH₂. In some embodiments of the compounds of formula III described above or below, X is O and A is CH-CR³R⁴-C(O)R². In some embodiments of the compounds of formula III described above or below, X is NH and A is CH-CR³R⁴-C(O)R²。

In some embodiments of the compounds of formula III or IIIa described above or below, R³And R⁴Are all hydrogen. In some embodiments of the compounds of formula III or IIIa described above or below, R³Is optionally substituted alkyl and R⁴Is hydrogen. In some embodiments of the compounds of formula III or IIIa described above or below, R³And R⁴Independently an optionally substituted alkyl group.

In some embodiments of the compounds of formula III or IIIa described above or below, R²Is an optionally substituted heteroaryl group. In certain embodiments, R²Is optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyridazinyl or optionally substituted pyrazinyl.

In some embodiments of the compounds of formula III or IIIa described above or below, R²Is phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, the substitutions on the phenyl groups are independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

In some embodiments described above or below of the compounds disclosed herein, B is CO₂R⁴And R is⁴Is an optionally substituted alkyl group. In some embodiments described above or below of the compounds disclosed herein, B is CO₂R⁴And R is⁴Is hydrogen.

In some embodiments described above or below of the compounds disclosed herein, n is 0, 1, or 2. In certain embodiments, n is 0. In certain embodiments, n is 1. In certain embodiments, R¹Independently selected from Cl, F, CH₂OH、CH₂NH₂、OCH₃、OCF₃、OCHF₂、CN、NO₂、CO₂H and CO₂CH₃。

In one aspect, provided herein is a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound selected from the group consisting of:

in another aspect, provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound selected from the group consisting of:

in some embodiments described above, the method is performed in vitro.

In some embodiments described above, the method is performed in vivo in a mammal, and the stem cell is present in the mammal. In some embodiments, the mammal is a domesticated animal or a farm animal. In certain embodiments, the mammal is a human, dog, cat, or horse.

In one aspect, provided herein is a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula Ia, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula Ib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula Ic, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula II, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula IIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula lib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula IIc, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound of formula III, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from:

in another aspect, provided herein is a compound, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, selected from the group consisting of:

in one aspect, provided herein is a pharmaceutical composition comprising a compound disclosed herein, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition further comprises an additional compound that is therapeutically effective for treating arthritis or joint damage and/or a symptom associated with arthritis or joint damage in a mammal. In certain embodiments, the additional compound is selected from NSAIDS, analgesics, angiopoietin-like 3 protein (ANGPTL3) or chondrogenic variants thereof, oral salmon calcitonin, SD-6010(iNOS inhibitor), vitamin D3 (cholecalciferol), apoptosis/caspase inhibitors (enrycan), collagen hydrolysates, FGF18, BMP7, Avocado Soy Unsaponifiables (ASU), and hyaluronic acid. In some embodiments, the mammal is a human. In other embodiments, the mammal is a companion animal or livestock. In further embodiments, the companion animal or livestock is a dog, cat, or horse.

In particular, the present invention relates to:

1. a method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula I:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴、CH₂CO₂H、CH₂CO₂R⁴Or optionally substituted phenyl;

X is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

Provided that

c) If Y is a bond and m is 0, then R²Selected from C (O) NR⁴R¹¹、(CR⁷R⁸)OR⁴、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴(ii) a And is

d) The compound is not selected from

2. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula Ia:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

X is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

3. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula Ib:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

R³Is H;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

4. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula Ic, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴；

Y is- (CR)⁵R⁶)-；

C is aryl or heteroaryl;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

5. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula I:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴、CH₂CO₂H、CH₂CO₂R³Or optionally substituted phenyl;

X is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

Provided that

b) The compound is not selected from

6. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula Ia:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

X is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

7. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula Ib:

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

R³Is H;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

8. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula Ic:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴；

Y is- (CR)⁵R⁶)-；

C is aryl or heteroaryl;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

9. The method of any of paragraphs 1,2, 5, or 6, wherein:

or R³To adjacent R³Or with R²Together forming a ring.

10. The method of paragraph 9 wherein:

Or R³To adjacent R³Or with R²Together forming a ring.

11. The method of paragraph 10, wherein R²Is F, Cl, C (O) CH₃、CH₃、CF₃、OCH₃、OEt、OPr、OCF₃、OCHF₂、CH₂OCH₃、CH₂OH、CH₂CH₂OH、CHOHCH₂OH、CH₂CH₂CH₂OH、CH₂CHOHCH₂OH、OCH₂C (O) OH or OCH₂C(O)NH₂。

12. The method of paragraph 11 wherein each R³Independently selected from CN, F, Cl, C (O) CH₃Or CO₂H。

13. The method of paragraph 12, wherein R²Is F, Cl, C (O) CH₃、CH₃、CF₃、OCH₃、OEt、OPr、OCF₃Or CH₂CH₂CH₂OH。

14. The method of any of paragraphs 1,3, 5, or 7, wherein:

R³Is H.

15. The method of paragraph 14 wherein R²Is (CR)⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹Or (CR)⁷R⁸)NR⁴SO₂R⁴。

16. The method of paragraph 15 wherein R²Is CH₂CH₂OH、CH₂CH₂OCH₃、CH₂CHCH₃OH、CHCH₃CH₂OH、CH₂CH₂CH₂OH、CH₂CH₂CH₂NH₂、CH₂CH₂CHCH₃OH、C(CH₃)₂CH₂CH₂OH、CH₂CH₂C(CH₃)₂OH、OCH₂CH₂OH、OCH₂CH₂OCH₃Or OCH₂CH₂NH₂。

17. The method of paragraph 14 wherein R²Is (CR)⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴Or X (CR)⁷R⁸)C(O)NR⁴R¹¹。

18. The method of paragraph 17 wherein R²Is CH₂C(O)CH₃、CH₂C(O)NH₂、CH₂CH₂C(O)CH₃Or CH₂CH₂C(O)NH₂。

19. The method of any of paragraphs 1,4, 5 or 8, wherein C is aryl.

20. The method of paragraph 19 wherein C is phenyl.

21. The method of any one of paragraphs 1,4, 5 or 8, wherein C is heteroaryl.

22. The method of paragraph 21, wherein C is pyridinyl, pyrimidinyl, pyridazinyl, or pyrazinyl.

23. The method of any of paragraphs 1,4, 5, 8 or 19-22, wherein:

Each R³Independently selected from H, CN, halo, CO₂H or haloalkyl.

24. The method as in paragraph 23 wherein:

Each R³Independently selected from H, CN, Cl, F, CO₂H or CF₃。

25. The method as recited in paragraph 24, wherein:

R²is Cl, F, C (O) CH₃、CO₂H、CH₃、OCH₃、CF₃(ii) a And is

Each R³Independently selected from H, CN or CO₂H。

26. The method of paragraph 24 wherein R²Is CH₂C(O)NH₂、CH₂C(O)CH₃、CH₂C(O)OH、CH₂CH₂C (O) OH or CH₂CH₂C(O)NH₂。

27. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula II:

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

provided that

a) If B is NHC (O) R²Or NR³C(O)R²Then A is not CO₂H; and is

b) The compound is not selected from

28. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula IIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is CO₂H or CO₂R³；

With the proviso that the compound is not

29. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula lib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

b is NHC (O) R²Or NR³C(O)R²；

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is SO₂NR^aR^b(ii) a And is

30. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula IIc:

wherein

n is 0, 1,2, 3 or 4;

A is C (O) NHR²Or C (O) NR²R⁴；

With the proviso that the compound is not

31. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula II:

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

provided that

a) If B is NHC (O) R²Or NR³C(O)R²Then A is not CO₂H; and is

b) The compound is not selected from

32. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula IIa:

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is CO₂H or CO₂R³；

With the proviso that the compound is not

33. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula lib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

b is NHC (O) R²Or NR³C(O)R²；

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is SO₂NR^aR^b(ii) a And is

34. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula IIc:

wherein

n is 0, 1,2, 3 or 4;

A is C (O) NHR²Or C (O) NR²R⁴；

With the proviso that the compound is not

35. The method of any one of paragraphs 27, 28, 31 or 32, wherein B is NHC (O) NHR²、NHC(O)NR²R⁴、NR³C(O)NHR²Or NR³C(O)NR²R⁴。

36. The method of paragraph 35, wherein B is NHC (O) NHR²Or NR³C(O)NHR²。

37. The method of paragraph 35, wherein B is NHC (O) NR²R⁴Or NR³C(O)NR²R⁴。

38. The method of paragraph 36 wherein B is NHC (O) NHR²。

39. The method of any of paragraphs 36-38, wherein R²Is optionally substituted phenyl.

40. The method of paragraph 39, wherein R²The phenyl group of (a) is disubstituted.

41. The method of paragraph 39, wherein R²The phenyl group of (a) is monosubstituted.

42. The method of paragraph 40 or 41, wherein R²The substitution on the phenyl group of (A) is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

43. The method of any one of paragraphs 27, 29, 31 or 33, wherein B is NHC (O) R²。

44. The method of any of paragraphs 27, 29, 31 or 33, wherein B is NR³C(O)R²。

45. The method of paragraph 44, wherein R³Is an optionally substituted alkyl group.

46. The method as in any one of paragraphs 43-45Process wherein each R^aAnd R^bIndependently an optionally substituted alkyl group.

47. The method of any of paragraphs 43-45, wherein R^aAnd R^bForm a ring together with the N to which they are attached.

48. The method of any of paragraphs 43-47, wherein R²Is optionally substituted phenyl.

49. The method of paragraph 48 wherein R²The phenyl group of (a) is disubstituted.

50. The method of paragraph 48 wherein R²The phenyl group of (a) is monosubstituted.

51. The method of paragraph 49 or 50, wherein R²The substitution on the phenyl group of (A) is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

52. The method of any of paragraphs 27, 30, 31 or 34, wherein B is NHSO₂R³、NR³SO₂R³、NHSO₂R⁴Or NR³SO₂R⁴。

53. The method of any of paragraphs 27, 30, 31 or 34, wherein B is NHSO₂NH₂、NHSO₂NHR²、NHSO₂NR²R⁴、NR³SO₂NH₂、NR³SO₂NHR²Or NR³SO₂NR²R⁴。

54. The method of paragraph 52, wherein B is NHSO₂R³Or NR³SO₂R³。

55. The method of paragraph 54 wherein B is NHSO₂R³。

56. Such as section 54 or55, wherein R³Is an optionally substituted alkyl group.

57. The method of paragraph 56 wherein R³Is CH₃。

58. The method of paragraph 52, wherein B is NHSO₂R⁴Or NR³SO₂R⁴。

59. The method of paragraph 58, wherein R⁴Is optionally substituted phenyl.

60. The method of any one of paragraphs 52-59, wherein A is C (O) NHR²。

61. The method of any of paragraphs 52-59, wherein A is C (O) NR²R⁴。

62. The method of paragraph 60 or 61, wherein R²Is optionally substituted phenyl.

63. The method of paragraph 62, wherein R²The phenyl group of (a) is disubstituted.

64. The method of paragraph 62, wherein R²The phenyl group of (a) is monosubstituted.

65. The method of paragraph 63 or 64 wherein R²The substitution on the phenyl group of (A) is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

66. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula III:

wherein

n is 0, 1,2, 3 or 4;

x is O, NH or NR⁶；

A is C (O), CH₂Or CH-CR³R⁴-C(O)R²；

R²Is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl;

R⁶Is optionally substituted phenyl;

provided that

d) If A is CH-CR³R⁴-C(O)R²X is O or NH;

e) if n is 0, A is CHCH₂C(O)R²And X is O, then R²Is not provided withAnd is

f) If A is C (O) or CH₂X is NR⁶And R is⁶Is not provided with

67. A method of ameliorating arthritis or joint damage in a mammal, the method comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound of formula IIIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

R²is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl; and is

68. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula III:

wherein

n is 0, 1,2, 3 or 4;

x is O, NH or NR⁶；

A is C (O), CH₂Or CH-CR³R⁴-C(O)R²；

R²Is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl;

R⁶Is optionally substituted phenyl;

provided that

a) If A is CH-CR³R⁴-C(O)R²X is O or NH;

b) if n is 0, A is CHCH₂C(O)R²And X is O, then R²Is not provided withAnd is

c) If A is C (O) or CH₂X is NR⁶And R is⁶Is not provided with

69. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound of formula IIIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof:

wherein

n is 0, 1,2, 3 or 4;

R²is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl; and is

70. The method of paragraph 66 or 68, wherein X is NR⁶And A is C (O).

71. The method of paragraph 66 or 68, wherein X is NR⁶And A is CH₂。

72. The method of paragraph 66 or 68, wherein X is O and A is CH-CR³R⁴-C(O)R²。

73. The method of paragraph 66 or 68, wherein X is NH and A is CH-CR³R⁴-C(O)R²。

74. The method of any of paragraphs 67, 69, 72 or 73, wherein R³And R⁴Are all hydrogen.

75. The method of any of paragraphs 67, 69, 72 or 73, wherein R³Is optionally substituted alkyl and R⁴Is hydrogen.

76. The method of any of paragraphs 67, 69, 72 or 73, wherein R³And R⁴Independently an optionally substituted alkyl group.

77. The method of any of paragraphs 74-76, wherein R²Is an optionally substituted heteroaryl group.

78. The method of any of paragraphs 74-76, wherein R²Is optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyridazinyl or optionally substituted pyrazinyl.

79. The method of any of paragraphs 74-76, wherein R²Is optionally substituted phenyl.

80. The method of any of paragraphs 79 wherein R²The phenyl group of (a) is disubstituted.

81. The method of any of paragraphs 79 wherein R²The phenyl group of (a) is monosubstituted.

82. The method of paragraph 80 or 81, wherein the substitution on the phenyl group is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂OH。

83. The method of any of paragraphs 1-65, wherein B is CO₂R⁴And R is⁴Is an optionally substituted alkyl group.

84. The method of any of paragraphs 1-65, wherein B is CO₂R⁴And R is⁴Is hydrogen.

85. The method of any of paragraphs 1-84, wherein n is 0, 1 or 2.

86. The method of paragraph 84, wherein n is 0.

87. The method of paragraph 84, wherein n is 1.

88. The method of paragraph 85 wherein R¹Independently selected from Cl, F, CH₂OH、CH₂NH₂、OCH₃、OCF₃、OCHF₂、CN、NO₂、CO₂H and CO₂CH₃。

89. The method of paragraph 1 or 5, wherein the compound is selected from

Or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

90. The method of paragraph 2 or 6, wherein said compound is selected from

91. The method of paragraph 3 or 7, wherein the compound is selected from

92. The method of paragraph 4 or 8, wherein the compound is selected from

93. The method of paragraph 27 or 31, wherein said compound is selected from

94. The method of paragraph 28 or 32, wherein said compound is selected from

95. The method of paragraph 29 or 33, wherein said compound is selected from

96. The method of paragraph 30 or 34, wherein said compound is selected from

97. The method of paragraph 66 or 68, wherein said compound is selected from

98. A method of ameliorating arthritis or joint damage in a mammal comprising administering to a joint of the mammal a composition comprising a therapeutically effective amount of a compound selected from the group consisting of

99. A method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound selected from the group consisting of

100. The method of any of paragraphs 5-8, 31-34, 68, 69, or 99, wherein the method is performed in vitro.

101. The method of any one of paragraphs 5-8, 31-34, 68, 69, or 99, wherein the method is performed in vivo in a mammal and the stem cells are present in the mammal.

102. The method of paragraph 101 wherein said mammal is a human, dog, cat or horse.

103. A compound of formula I selected from

104. A compound of formula Ia selected from

105. A compound of formula Ib, selected from

106. A compound of formula Ic selected from

107. A compound of formula II selected from

108. A compound of formula IIa selected from

109. A compound of formula IIb selected from

110. A compound of formula IIc selected from

111. A compound of formula III selected from

112. A compound selected from

113. A pharmaceutical composition comprising a compound as described in any one of paragraphs 103-112 or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, and a pharmaceutically acceptable excipient.

114. The pharmaceutical composition of paragraph 113, further comprising an additional compound that is therapeutically effective for treating arthritis or joint damage and/or a symptom associated with arthritis or joint damage in a mammal.

115. The pharmaceutical composition of paragraph 114, wherein the additional compound is selected from NSAIDS, analgesics, angiopoietin-like 3 protein (ANGPTL3) or chondrogenic variants thereof, oral salmon calcitonin, iNOS inhibitors, vitamin D3, caspase inhibitors, collagen hydrolysates, FGF18, BMP7, Avocado Soybean Unsaponifiables (ASU), and hyaluronic acid.

116. The pharmaceutical composition of paragraph 114, wherein said mammal is a human.

117. The pharmaceutical composition of paragraph 114 wherein said mammal is a companion animal or livestock.

118. The method of any one of paragraphs 1-4, 27-30, 66, 67 or 98, wherein the mammal is a human.

119. The method of any of paragraphs 1-4, 27-30, 66, 67 or 98, wherein said mammal is a domesticated or livestock animal.

Is incorporated by reference

All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

Detailed Description

Osteoarthritis (OA) is characterized by progressive breakdown of articular cartilage and ultimately leads to dysfunction of synovial joints [ Reginster, j.y. and n.g. khaltaev, Introduction and WHO perspective on the spherical burden of mucosulques regulation (Oxford),2002.41Supp1: p.1-2 ]. OA is mediated by several pathogenesis mechanisms, including enzymatic degradation of the extracellular matrix, defective new matrix formation, cell death, and abnormal activation and hypertrophic differentiation of chondrocytes [ Goldring, m.b. and s.r.goldring, articular and subendoral bone in the pathogenesis of osteopathoritis, ann N YAcad Sci,2010.1192(1): p.230-7 ]. The only treatment options currently available for OA are pain management and surgical intervention [ Hunter, d.j., pharmacological therapy for osteo-the era of disease modification. nat Rev rheumato, 2011.7(1): p.13-22 ].

Mesenchymal Stem Cells (MSCs) present in bone marrow and most adult tissues are capable of self-renewal and differentiation into a variety of cell lineages, including chondrocytes, osteoblasts and adipocytes [ Pittenger, m.f. et al, multilineagent potential of adult human mesenchymal cells, science,1999.284(5411): p.143-7 ]. Recent studies have found that adult articular cartilage contains MSCs (about 3% of cells) capable of multi-lineage differentiation. In OA cartilage, the number of these cells roughly doubles. These existing stem cells still retain the ability to differentiate into chondrocytes, and thus retain the ability to repair damaged cartilage [ Grogan, S.P. et al, sensory promoter cell markers in human tissue, normal distribution and transformation in osteo-arthritis Res Ther,2009.11(3): p.R85; koelling, S. et al, Migratorychondrogen genetic reagent cells from repair tissue reducing the later stages of human osteo-analysis Cell 2009.4(4): p.324-35 ].

The present invention is based in part on the following findings: the compounds of the invention stimulate chondrocyte differentiation in mesenchymal stem cells. Accordingly, the present invention provides a method of inducing differentiation of mesenchymal stem cells into chondrocytes. Further, the present invention provides administering the compounds and compositions of the present invention to prevent or ameliorate arthritis or joint injury by administering the compounds or compositions of the present invention to the joint, spine, intervertebral disc, or systemically.

Definition of

In the following description, certain specific details are set forth in order to provide a thorough understanding of the embodiments. However, it will be understood by those skilled in the art that the present invention may be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring descriptions of the embodiments. Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be construed in an open, inclusive sense, i.e., as "including but not limited to". Further, the headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed invention.

Reference throughout the specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments. In addition, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise. It should also be noted that the term "or" is generally employed in its sense including "and/or" unless the content clearly dictates otherwise.

As used herein, unless otherwise indicated, the following terms have the following meanings:

"amino" means-NH₂A group.

"cyano" or "nitrile" refers to the group-CN.

"Hydroxy or hydroxyl" refers to the-OH group.

"nitro" means-NO₂A group.

"oxo" refers to an ═ O substituent.

"oxime" refers to the N-OH substituent.

"thio" refers to ═ S substituents.

"alkyl" refers to a straight or branched hydrocarbon chain radical group, either fully saturated or containing unsaturation, having from 1 to 30 carbon atoms, and attached to the rest of the molecule by a single bond. Alkyl groups containing any number of carbon atoms from 1 to 30 are contemplated. Alkyl groups containing up to 30 carbon atoms are designated C₁-C₃₀Alkyl, likewise, for example, alkyl containing up to 12 carbon atoms is C₁-C₁₂An alkyl group. Alkyl groups containing other numbers of carbon atoms (and other moieties as defined herein) are represented in a similar manner. Alkyl groups include, but are not limited to, C₁-C₃₀Alkyl radical, C₁-C₂₀Alkyl radical, C₁-C₁₅Alkyl radical, C₁-C₁₀Alkyl radical, C₁-C₈Alkyl radical, C₁-C₆Alkyl radical, C₁-C₄Alkyl radical, C₁-C₃Alkyl radical, C₁-C₂Alkyl radical, C₂-C₈Alkyl radical, C₃-C₈Alkyl and C₄-C₈An alkyl group. Representative alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, isobutyl, sec-butyl, n-pentyl, 1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, vinyl, allyl, propynyl, and the like. The unsaturated bond-containing alkyl group includes an alkenyl group and an alkynyl group. Unless otherwise specifically stated in the specification, alkyl groups may be optionally substituted as described below.

"alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain as described above for alkyl groups. Unless otherwise specifically stated in the specification, the alkylene group may be optionally substituted as described below.

"alkoxy" means a group of the formula-OR_aWherein R is_aIs an alkyl group as defined above. Unless otherwise specifically stated in the specification, alkoxy groups may be optionally substituted as described below.

"aryl" refers to a group derived from a hydrocarbon ring system comprising hydrogen, 6 to 30 carbon atoms, and at least one aromatic ring. The aryl group may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, and may include fused or bridged ring systems. Aryl groups include, but are not limited to, the group consisting of aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, and mixtures thereof,Fluoranthene, fluorene, asymmetric indacene (as-indacene), symmetric indacene (s-indacene), indane, indene, naphthalene, phenalene, phenanthrene, obsidian, pyrene and pyreneAryl groups derived from hydrocarbon ring systems of triphenylene. Unless otherwise specifically stated in the specification, the term "aryl" or the prefix "aryl" (as in "aralkyl") is intended to include optionally substituted aryl groups.

"cycloalkyl" or "carbocycle" refers to a stable, non-aromatic, monocyclic or polycyclic, carbocyclic ring which may include fused or bridged ring systems, which may be saturated or unsaturated. Representative cycloalkyl or carbocycle groups include, but are not limited to, cycloalkyl groups having 3 to 15 carbon atoms, 3 to 10 carbon atoms, 3 to 8 carbon atoms, 3 to 6 carbon atoms, 3 to 5 carbon atoms, or 3 to 4 carbon atoms. Monocyclic cycloalkyl or carbocycle includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Polycyclic groups cycloalkyl or carbocycle include, for example, adamantyl, norbornyl, decahydronaphthyl, bicyclo [3.3.0] octane, bicyclo [4.3.0] nonane, cis-decahydronaphthalene, trans-decahydronaphthalene, bicyclo [2.1.1] hexane, bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane, bicyclo [3.2.2] nonane and bicyclo [3.3.2] decane and 7, 7-dimethyl-bicyclo [2.2.1] heptanyl. Unless specifically stated otherwise in the specification, the cycloalkyl or carbocycle may be optionally substituted. Illustrative examples of cycloalkyl groups include, but are not limited to, the following moieties:

and the like.

"fused" refers to any of the ring structures described herein that are fused to the ring structure present. When the fused ring is a heterocyclyl or heteroaryl ring, any carbon atom on the existing ring structure that is part of the fused heterocyclyl or heteroaryl ring may be replaced by a nitrogen atom.

"halo (halo)" or "halogen" refers to bromo, chloro, fluoro or iodo.

"haloalkyl" refers to an alkyl group as defined above substituted with one or more halo groups as defined above, e.g., trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2, 2-trifluoroethyl, 1, 2-difluoroethyl, 3-bromo-2-fluoropropyl, 1, 2-dibromoethyl, and the like. Unless otherwise specifically stated in the specification, haloalkyl may be optionally substituted.

Similarly, "haloalkoxy" refers to the formula-OR_aWherein R is_aIs haloalkyl as defined above. Unless otherwise specifically stated in the specification, haloalkoxy groups may be optionally substituted as described below.

"heterocycloalkyl" or "heterocyclyl" or "heterocyclic ring" or "heterocycle" refers to a stable 3 to 24-membered non-aromatic ring group containing 2 to 23 carbon atoms and 1 to 8 heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur. Unless otherwise specifically stated in the specification, the heterocyclic group may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, and may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heterocyclic group may be optionally oxidized; the nitrogen atoms may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. Examples of such heterocyclyl groups include, but are not limited to, azetidinyl, dioxolanyl, thienyl [1,3] dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidinonyl (4-piperidonyl), pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, 1-dioxo-thiomorpholinyl, 12-crown-4, 15-crown-5-ether, 18-crown-6, 21-crown-7, aza-18-crown-6, diaza-18-crown-6, aza-21-crown-7 and diaza-21-crown-7. Unless specifically stated otherwise in the specification, the heterocyclic group may be optionally substituted. Illustrative examples of heterocycloalkyl groups, also referred to as non-aromatic heterocycles, include:

and the like. The term heterocycloalkyl also includes carbohydrates in all ring forms, including but not limited to monosaccharides, disaccharides, and oligosaccharides. Unless otherwise specified, heterocycloalkyl groups have 2 to 10 carbons in the ring. It is understood that when referring to the number of carbon atoms in a heterocycloalkyl group, the number of carbon atoms in the heterocycloalkyl group is different from the total number of atoms (including heteroatoms) that make up the heterocycloalkyl group (i.e., the backbone atoms of the heterocycloalkyl ring). Unless specifically stated otherwise in the specification, heterocycloalkyl groups may be optionally substituted.

"heteroaryl" refers to a 5 to 14 membered ring system group containing hydrogen atoms, 1 to 13 carbon atoms, 1 to 6 heteroatoms selected from nitrogen, oxygen, phosphorus and sulfur, and at least one aromatic ring. For purposes of this invention, heteroaryl groups can be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, and can include fused or bridged ring systems; and the nitrogen, carbon or sulfur atom in the heteroaryl group may be optionally oxidized; the nitrogen atoms may optionally be quaternized. Examples include, but are not limited to, azaA group selected from the group consisting of acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo [ b][1,4]Dioxa medicineA group, a1, 4-benzodioxanyl group, a benzonaphthofuranyl group, a benzoxazolyl group, a benzodioxolyl group, a benzodioxinyl group, a benzopyranyl group, a benzonaphthofuranyl group, a benzoxaxolyl group, a benzodioxinyl group, a benzofuranyl group, a benzoPyronyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo [4,6 ]]Imidazo [1,2-a ]]Pyridyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothienyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxo-aza-azanylA group, an oxazolyl group, an oxiranyl group, a 1-oxopyridyl group, a 1-oxopyrimidinyl group, a 1-oxopyridazinyl group, a 1-phenyl-1H-pyrrolyl group, a phenazinyl group, a phenothiazinyl group, a phenoxazinyl group, a phthalazinyl group, a pteridinyl group, a purinyl group, a pyrrolyl group, a pyrazolyl group, a pyridyl group, a pyrazinyl group, a pyrimidinyl group, a pyridazinyl group, a quinazolinyl group, a quinoxalinyl group, a quinolyl group, a quinuclidinyl group, an isoquinolyl group, a tetrahydroquinolyl group, a thiazolyl group, a thiadiazolyl group, a triazolyl group, a tetrazolyl group, a triazinyl group, and a thienyl group (thiophenyl) (i.e. Unless specifically stated otherwise in the specification, heteroaryl groups may be optionally substituted.

All of the above groups may be substituted or unsubstituted. The term "substituted" as used herein means that any of the above groups (e.g., alkyl, alkylene, alkoxy, aryl, cycloalkyl, haloalkyl, heterocyclyl, and/or heteroaryl) can be further functionalized wherein at least one hydrogen atom is replaced by a bond to a substituent other than a hydrogen atom. Unless specifically stated in the specification, substituted groups may include one or more substituents selected from: oxo, amino, -CO₂H. Nitrile, nitro, hydroxy, thioxo (thioxy), alkyl, alkylene, alkoxy, aryl, cycloalkyl, heterocyclyl, heteroaryl, dialkylamine, arylamine, alkylarylamine, diarylamine, trialkylammonium (-N)⁺R₃) N-oxides, imides and enamines; in a solvent such as trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, triarylsilylA perfluoroalkyl group or a perfluoroalkoxy group, such as a trifluoromethyl group or a trifluoromethoxy group. "substituted" also refers to any of the foregoing groups in which one or more hydrogen atoms are replaced by a higher order bond (e.g., a double or triple bond) to a heteroatom, such as oxygen in oxo, carbonyl, carboxyl, and ester groups, and nitrogen in groups such as imines, oximes, hydrazones, and nitriles. For example, "substituted" includes where one or more hydrogen atoms are replaced by-NH₂、-NR_gC(＝O)NR_gR_h、-NR_gC(＝O)OR_h、-NR_gSO₂R_h、-OC(＝O)NR_gR_h、-OR_g、-SR_g、-SOR_g、-SO₂R_g、-OSO₂R_g、-SO₂OR_g、＝NSO₂R_gand-SO₂NR_gR_hSubstituted any of the above groups. In the above, R_gAnd R_hThe same or different, and are independently hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and/or heteroarylalkyl. Further, each of the above substituents may be optionally substituted with one or more of the above substituents. In addition, any of the above groups may be substituted to include one or more internal oxygen, sulfur or nitrogen atoms. For example, an alkyl group may be substituted with one or more internal oxygen atoms to form an ether or polyether group. Similarly, alkyl groups may be substituted with one or more internal sulfur atoms to form thioethers, disulfides, and the like.

The term "optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, "optionally substituted alkyl" means "alkyl" or "substituted alkyl" as defined above. Further, the optionally substituted group may be unsubstituted (e.g., -CH)₂CH₃)、Fully substituted (e.g., -CF)₂CF₃) Monosubstituted (e.g., -CH)₂CH₂F) Or substituted at any level between fully and mono-substituted (e.g., -CH)₂CHF₂、-CH₂CF₃、-CF₂CH₃、-CFHCHF₂Etc.). It will be understood by those skilled in the art that, for any group containing one or more substituents, these groups are not intended to introduce any substitution or substitution pattern that is not sterically impractical and/or synthetically non-feasible (e.g., substituted alkyl includes optionally substituted cycloalkyl, which in turn is defined as including optionally substituted alkyl, as such may be recycled indefinitely). Thus, any substituent group described is generally understood to have a maximum molecular weight of about 1,000 daltons, and more typically, up to about 500 daltons.

An "effective amount" or "therapeutically effective amount" refers to the amount of a compound that is administered to a mammalian subject as a single dose or as part of a series of doses and is effective to produce the desired therapeutic effect.

"treatment" of an individual (e.g., a mammal such as a human) or a cell is any type of intervention used in an attempt to alter the natural course of the individual or cell. In some embodiments, treatment includes administration of the pharmaceutical composition after the onset of a pathological event or contact with a pathogen, and includes stabilization of the condition (e.g., the condition does not worsen) or alleviation of the condition. In other embodiments, treatment also includes prophylactic treatment (e.g., administration of a composition described herein when the subject is suspected of having a bacterial infection).

"tautomer" refers to the transfer of a proton from one atom of a molecule to another atom of the same molecule. The compounds provided herein can exist as tautomers. Tautomers are compounds that can interconvert by the migration of a hydrogen atom (with the conversion of a single bond and an adjacent double bond). In a bonding arrangement where tautomerism is likely to occur, there will be a chemical equilibrium of the tautomers. All tautomeric forms of the compounds disclosed herein are contemplated. The exact ratio of tautomers depends on several factors including temperature, solvent and pH. Some examples of interconversion of tautomers include:

a "metabolite" of a compound disclosed herein is a derivative of the compound that is formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound that is formed when the compound is metabolized. As used herein, the term "metabolism" refers to the sum of processes (including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes, such as oxidation reactions) by which a particular substance is altered by an organism. Thus, enzymes may produce specific structural changes to a compound. For example, cytochrome P450 catalyzes a variety of oxidation and reduction reactions, while uridine diphosphate glucuronyl transferase catalyzes the conversion of an activated glucuronic acid molecule to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines and free thiols. Further information on metabolism is available from the pharmacological Basis of Therapeutics, ninth edition, McGraw-Hill (1996). Metabolites of the compounds disclosed herein can be identified by the following methods: administering the compound to a host and analyzing a tissue sample from the host, or incubating the compound with hepatocytes in vitro and analyzing the resulting compound. Both of these methods are well known in the art. In some embodiments, a metabolite of a compound is formed by an oxidative process and corresponds to a corresponding hydroxyl-containing compound. In some embodiments, the compound is metabolized to a pharmacologically active metabolite.

Method of producing a composite material

Provided herein is a method of inducing differentiation of mesenchymal stem cells into chondrocytes, the method comprising contacting mesenchymal stem cells with a sufficient amount of a compound disclosed herein, thereby inducing differentiation of stem cells into chondrocytes.

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴、CH₂CO₂H、CH₂CO₂R⁴Or optionally substituted phenyl;

X is O or CR⁵R⁶；

Or R³To adjacent R³Or with R²Together form a ring;

each R⁴Independently selected from H and optionally substituted alkyl;

Provided that

b) The compound is not selected from

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

Or R³To adjacent R³Or with R²Together form a ring;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

R³Is H;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴；

Y is- (CR)⁵R⁶)-；

C is aryl or heteroaryl;

x is O or CR⁵R⁶；

Or R³To adjacent R³Or with R²Together form a ring;

each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴、CH₂CO₂H、CH₂CO₂R³Or optionally substituted phenyl;

X is O or CR⁵R⁶；

Or R³To adjacent R³Or with R²Together form a ring;

each R⁴Independently selected from H and optionally substituted alkyl;

Provided that

R²Is not provided withC(O)NH₂、p-CH₂OR⁴、p-CH(OH)CH₂OH、p-CH₂CH₂OH or p-CH₂CH₂CH₂OH; and is

b) The compound is not selected from

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

Or R³To adjacent R³Or with R²Together form a ring;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

wherein

n is 0, 1,2, 3 or 4;

b is CO₂R⁴；

R³Is H;

x is O or CR⁵R⁶；

Each R⁴Independently selected from H and optionally substituted alkyl;

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴；

Y is- (CR)⁵R⁶)-；

C is aryl or heteroaryl;

x is O or CR⁵R⁶；

R²Is halo, C (O) R⁴、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, alkyl halideBase, SO₂R⁴、SO₂NH₂、SO₃H、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴Or C (═ NOR)⁴)R⁴；

Or R³To adjacent R³Or with R²Together form a ring;

each R⁴Independently selected from H and optionally substituted alkyl;

With the proviso that the compound is not selected from

In some embodiments described above or below for compounds of formula I or Ia:

or R³To adjacent R³Or with R²Together forming a ring.

Or R³To adjacent R³Or with R²Together forming a ring.

In certain embodiments, R³Independently selected from CN, F, Cl, C (O) CH₃Or CO₂H. In certain embodiments, R³Is CN or CO₂H. In certain embodiments, R²Is F, Cl, C (O) CH₃、CH₃、CF₃、OCH₃、OEt、OPr、OCF₃Or CH₂CH₂CH₂And (5) OH. In certain embodiments, R²Is CH₂CH₂CH₂And (5) OH. In certain embodiments, R³To adjacent R³Or with R²Together areForming a ring.

In certain embodiments, R²Is (CR)⁷R⁸)OR⁴、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)C(O)OR⁴Or X (CR)⁷R⁸)C(O)NR⁴R¹¹(ii) a And each R³Independently selected from CN, F, Cl, C (O) CH₃、CO₂H、C(O)NH₂、CH₃、OCF₃Or OCH₃. In certain embodiments, R²Is F, Cl, C (O) CH₃、CH₃、CF₃、OCH₃、OEt、OPr、OCF₃Or CH₂CH₂CH₂OH; and R is³Independently selected from CN, F, Cl, C (O) CH₃Or CO₂H. In certain embodiments, R²Is F, Cl, C (O) CH₃、CH₃、CF₃、OCH₃、OEt、OPr、OCF₃Or CH₂CH₂CH₂OH; and R is³Independently selected from CN or CO₂H. In certain embodiments, R²Is CH₂CH₂CH₂OH and R³Independently selected from CN, F, Cl, C (O) CH₃Or CO₂H。

In some embodiments described above or below for compounds of formula I:

R²is halo, C (O) R⁴、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴Or C (═ NOR)⁴)R⁴(ii) a And is

Each R³Independently selected from CN, halo, C (O) R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴。

In some embodiments described above or below for compounds of formula Ia:

In some embodiments described above or below for compounds of formula Ib:

R³Is H.

In some embodiments described above or below for compounds of formula Ic, C is aryl. In certain embodiments, C is phenyl. In certain embodiments, C is naphthyl.

In some embodiments described above or below for compounds of formula Ic, C is heteroaryl. In certain embodiments, C is pyridinyl, pyrimidinyl, pyridazinyl, or pyrazinyl. In certain embodiments, C is pyridyl. In certain embodiments, C is pyrimidinyl. In certain embodiments, C is pyridazinyl. In certain embodiments, C is a 5 membered heteroaryl ring. In certain embodiments, C is thiophene, benzofuran, pyrrole, thiazole, imidazole, oxazole, pyrazole, or triazole.

In some embodiments described above or below for compounds of formula Ic:

R²is halo, C (O) R⁴、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂R⁴、SO₂NH₂、SO₃H、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴Or C (═ NOR)⁴)R⁴(ii) a And is

Each R³Independently selected from H, CN, halo, C (O) R⁴、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy, haloalkyl, SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴；

Provided that if n ═ 0 and C is phenyl, R²Not CO₂H or p-OCH₃。

In some embodiments described above or below for compounds of formula Ic:

Each R³Independently selected from H, CN, halo, CO₂H or haloalkyl.

In certain embodiments described above or below for compounds of formula Ic:

Each R³Independently selected from H, CN, Cl, F, CO₂H or CF₃。

In certain embodiments, R²Is Cl, F, C (O) CH₃、CO₂H、CH₃、OCH₃、CF₃(ii) a And each R³Independently selected from H, CN or CO₂H. In certain embodiments, R²Is CH₂C(O)NH₂、CH₂C(O)CH₃、CH₂C(O)OH、CH₂CH₂C (O) OH or CH₂CH₂C(O)NH₂. In certain embodiments, R²Is CO₂H. In certain embodiments, R²Is CO₂H and each R³Independently selected from H, CN, Cl, F or CF₃。

In certain embodiments described above or below for compounds of formula Ic:

Each R³Independently selected from H, CN or CO₂H。

In certain embodiments, R²Is CH₂C(O)NH₂、CH₂C(O)CH₃、CH₂C(O)OH、CH₂CH₂C (O) OH or CH₂CH₂C(O)NH₂(ii) a And each R³Independently selected from H, CN or CO₂H。

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

R⁵is H, renOptionally substituted alkyl, C (O) R⁴、C(O)OR⁴、C(O)NR⁴R⁴Or SO₂R⁴；

provided that

a) If B is NHC (O) R²Or NR³C(O)R²Then A is not CO₂H; and is

b) The compound is not selected from

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is CO₂H or CO₂R³；

With the proviso that the compound is not

wherein

n is 0, 1,2, 3 or 4;

b is NHC (O) R²Or NR³C(O)R²；

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is SO₂NR^aR^b(ii) a And is

wherein

n is 0, 1,2, 3 or 4;

A is C (O) NHR²Or C (O) NR²R⁴；

With the proviso that the compound is not

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

provided that

a) If B is NHC (O) R²Or NR³C(O)R²Then A is not CO₂H; and is

b) The compound is not selected from

wherein

n is 0, 1,2, 3 or 4;

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is CO₂H or CO₂R³；

With the proviso that the compound is not

wherein

n is 0, 1,2, 3 or 4;

b is NHC (O) R²Or NR³C(O)R²；

R³is optionally substituted alkyl or optionally substituted aralkyl;

A is SO₂NR^aR^b(ii) a And is

wherein

n is 0, 1,2, 3 or 4;

A is C (O) NHR²Or C (O) NR²R⁴；

With the proviso that the compound is not

In some embodiments of the compounds of formula IIa described above or below, B is NHC (O) NHR²、NHC(O)NR²R⁴、NR³C(O)NHR²Or NR³C(O)NR²R⁴. In certain embodiments, B is NHC (O) NHR²Or NR³C(O)NHR². In certain embodiments, B is NHC (O) NR²R⁴Or NR³C(O)NR²R⁴. In certain embodiments, B is NHC (O) NHR²。

In some embodiments described above or below for compounds of formula IIaB is NHC (O) NHR²、NHC(O)NR²R⁴、NR³C(O)NHR²Or NR³C(O)NR²R⁴(ii) a And A is CO₂H. In certain embodiments, B is NHC (O) NHR²Or NR³C(O)NHR²(ii) a And A is CO₂H. In certain embodiments, B is NHC (O) NHR²And A is CO₂H. In certain embodiments, B is NHC (O) NHR²And A is CO₂H, wherein R²Is optionally substituted phenyl.

In some embodiments of the compounds of formula IIa described above or below, B is NHC (O) NHR²、NHC(O)NR²R⁴、NR³C(O)NHR²Or NR³C(O)NR²R⁴(ii) a And A is CO₂R³. In certain embodiments, B is NHC (O) NHR²Or NR³C(O)NHR²(ii) a And A is CO₂R³. In certain embodiments, B is NHC (O) NHR²And A is CO₂R³. In certain embodiments, B is NHC (O) NHR²And A is CO₂R³Wherein R is²Is optionally substituted phenyl.

In some embodiments of the compounds of formula IIa, described above or below, R²Is optionally substituted phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, R²The substitution on the phenyl group of (a) is independently selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted aryloxy, halo, CN, CO₂H. Amino, monoalkylamine, dialkylamine, monoarylamine, alkylarylamine, cycloalkyl, hydroxy, C (O) - (optionally substituted alkyl), C (O) NH₂C (O) NH- (optionally substituted alkyl), alkyl sulfide, alkyl sulfoxide, alkyl sulfone, C (O) - (optionally substituted aryl), C (O) NH- (optionally substituted aryl), aryl sulfide, aryl sulfoxide or aryl sulfone. In certain embodiments, R²Is independently selected from F, Cl、CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂And (5) OH. In certain embodiments, R²Is disubstituted by CN and is selected from F, Cl, CO₂H、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²By disubstituted radicals of CO on the phenyl radical of₂H and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²Disubstituted on the phenyl radical of (A) by CH₂CH₂CH₂OH and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CO₂The radical composition of H.

In some embodiments of the compounds of formula IIa, described above or below, R²Is optionally substituted naphthyl.

In some embodiments of the compounds of formula IIa, described above or below, R²Is an optionally substituted heteroaryl group. In certain embodiments, R²Is optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyridazinyl or optionally substituted pyrazinyl. In certain embodiments, R²Is an optionally substituted 5-membered heteroaryl ring. In certain embodiments, the 5-membered heteroaryl ring is thiophene, benzofuran, pyrrole, thiazole, imidazole, oxazole, pyrazole, or triazole. In certain embodiments, R²Is an optionally substituted bicyclic heteroaryl. In certain embodiments, the bicyclic heteroaryl is benzimidazole, benzothiazole, benzoxazole, indazole, quinoline, or naphthyridine.

Above the compound of formula IIb orIn some embodiments described below, B is NHC (O) R². In certain embodiments, B is NHC (O) R²And R is²Is optionally substituted phenyl. In certain embodiments, B is NHC (O) R²And R is²Is an optionally substituted heteroaryl group.

In some embodiments of the compounds of formula IIb described above or below, each R^aAnd R^bIndependently an optionally substituted alkyl group. In certain embodiments, each R is^aAnd R^bIndependently an alkyl group. In some embodiments of the compounds of formula IIb described above or below, R^aAnd R^bForm a ring together with the N to which they are attached. In certain embodiments, the ring is morpholinyl, thiomorpholinyl, piperidinyl, pyrrolidinyl, azetidinyl, aziridinyl, azepanyl, homopiperazinyl, or piperazinyl.

In some embodiments of the compounds of formula IIb described above or below, R²Is optionally substituted phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, R²The substitution on the phenyl group of (a) is independently selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted aryloxy, halo, CN, CO₂H. Amino, monoalkylamine, dialkylamine, monoarylamine, alkylarylamine, cycloalkyl, hydroxy, C (O) - (optionally substituted alkyl), C (O) NH₂C (O) NH- (optionally substituted alkyl), alkyl sulfide, alkyl sulfoxide, alkyl sulfone, C (O) - (optionally substituted aryl), C (O) NH- (optionally substituted aryl), aryl sulfide, aryl sulfoxide or aryl sulfone. In certain embodiments, R²The substitution on the phenyl group of (A) is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂And (5) OH. In certain embodiments, R²Is disubstituted by CN and is selected from F, Cl, CO₂H、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²By disubstituted radicals of CO on the phenyl radical of₂H and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²Disubstituted on the phenyl radical of (A) by CH₂CH₂CH₂OH and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CO₂The radical composition of H.

In some embodiments of the compounds of formula IIb described above or below, R²Is optionally substituted naphthyl.

In some embodiments of the compounds of formula IIb described above or below, R²Is an optionally substituted heteroaryl group. In certain embodiments, R²Is optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyridazinyl or optionally substituted pyrazinyl. In certain embodiments, R²Is an optionally substituted 5-membered heteroaryl ring. In certain embodiments, the 5-membered heteroaryl ring is thiophene, benzofuran, pyrrole, thiazole, imidazole, oxazole, pyrazole, or triazole. In certain embodiments, R²Is an optionally substituted bicyclic heteroaryl. In certain embodiments, the bicyclic heteroaryl is benzimidazole, benzothiazole, benzoxazole, indazole, quinoline, or naphthyridine.

In some embodiments of the compounds of formula IIc described above or below, B is NHSO₂R³、NR³SO₂R³、NHSO₂R⁴Or NR³SO₂R⁴. In certain embodiments, B is NHSO₂R³Or NR³SO₂R³. In certain embodiments, B is NHSO₂R³. In certain embodiments, R³Is an optionally substituted alkyl group. In certain embodiments, R³Is an alkyl group. In certain embodiments, R³Is CH₃. In certain embodiments, B is NHSO₂R⁴Or NR³SO₂R⁴. In certain embodiments, R⁴Is optionally substituted phenyl. In certain embodiments, R⁴Is optionally substituted naphthyl. In certain embodiments, R⁴Is an optionally substituted heteroaryl group. In certain embodiments, R⁴Is an optionally substituted heterocyclic group.

In some embodiments of the compounds of formula IIc described above or below, B is NHSO₂R³、NR³SO₂R³、NHSO₂R⁴Or NR³SO₂R⁴And A is C (O) NHR². In certain embodiments, B is NHSO₂R³Or NR³SO₂R³And A is C (O) NHR². In certain embodiments, B is NHSO₂R³And A is C (O) NHR². In certain embodiments, B is NHSO₂R³And A is C (O) NHR²Wherein R is³Is an optionally substituted alkyl group. In certain embodiments, B is NHSO₂R³And A is C (O) NHR²Wherein R is³Is optionally substituted alkyl and R²Is optionally substituted phenyl. In certain embodiments, B is NHSO₂R³And A is C (O) NHR²Wherein R is³Is optionally substituted alkyl and R²Is an optionally substituted heteroaryl group.

In some embodiments of the compounds of formula IIc described above or below, B is NHSO₂R³、NR³SO₂R³、NHSO₂R⁴Or NR³SO₂R⁴And A is C (O) NR²R⁴. In certain embodiments, B is NHSO₂R³Or NR³SO₂R³And A is C (O) NR²R⁴. In certain embodiments, B is NHSO₂R³And A is C (O) NR²R⁴. In certain embodiments, B is NHSO₂R³And A is C (O) NR²R⁴Wherein R is³Is an optionally substituted alkyl group. In certain embodiments, B is NHSO₂R³And A is C (O) NR²R⁴Wherein R is³Is optionally substituted alkyl and R²Is optionally substituted phenyl. In certain embodiments, B is NHSO₂R³And A is C (O) NR²R⁴Wherein R is³Is optionally substituted alkyl and R²Is an optionally substituted heteroaryl group.

In some embodiments of the compounds of formula IIc described above or below, B is NHSO₂NH₂、NHSO₂NHR²、NHSO₂NR²R⁴、NR³SO₂NH₂、NR³SO₂NHR²Or NR³SO₂NR²R⁴. In certain embodiments, B is NHSO₂NH₂、NHSO₂NHR²、NHSO₂NR²R⁴、NR³SO₂NH₂、NR³SO₂NHR²Or NR³SO₂NR²R⁴And A is C (O) NHR². In certain embodiments, B is NHSO₂NH₂、NHSO₂NHR²、NHSO₂NR²R⁴、NR³SO₂NH₂、NR³SO₂NHR²Or NR³SO₂NR²R⁴And A is C (O) NR²R⁴。

Some of the compounds described above or below for formula IIcIn embodiments, A is C (O) NHR². In some embodiments of the compounds of formula IIc described above or below, A is C (O) NR²R⁴. In certain embodiments, R²Is optionally substituted phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, R²The substitution on the phenyl group of (a) is independently selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted aryloxy, halo, CN, CO₂H. Amino, monoalkylamine, dialkylamine, monoarylamine, alkylarylamine, cycloalkyl, hydroxy, C (O) - (optionally substituted alkyl), C (O) NH₂C (O) NH- (optionally substituted alkyl), alkyl sulfide, alkyl sulfoxide, alkyl sulfone, C (O) - (optionally substituted aryl), C (O) NH- (optionally substituted aryl), aryl sulfide, aryl sulfoxide or aryl sulfone. In certain embodiments, R²The substitution on the phenyl group of (A) is independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂And (5) OH. In certain embodiments, R²Is disubstituted by CN and is selected from F, Cl, CO₂H、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²By disubstituted radicals of CO on the phenyl radical of₂H and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²Disubstituted on the phenyl radical of (A) by CH₂CH₂CH₂OH and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CO₂The radical composition of H. In certain embodiments, R²Is optionally substituted naphthyl.

In some embodiments of the compounds of formula IIc described above or below, R²Is an optionally substituted heteroaryl group. In certain embodiments, R²Is optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyridazinyl or optionally substituted pyrazinyl. In certain embodiments, R²Is an optionally substituted 5-membered heteroaryl ring. In certain embodiments, the 5-membered heteroaryl ring is thiophene, benzofuran, pyrrole, thiazole, imidazole, oxazole, pyrazole, or triazole. In certain embodiments, R²Is an optionally substituted bicyclic heteroaryl. In certain embodiments, the bicyclic heteroaryl is benzimidazole, benzothiazole, benzoxazole, indazole, quinoline, or naphthyridine.

wherein

n is 0, 1,2, 3 or 4;

x is O, NH or NR⁶；

A is C (O), CH₂Or CH-CR³R⁴-C(O)R²；

R²Is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl;

R⁶Is optionally substituted phenyl;

provided that

a) If A is CH-CR³R⁴-C(O)R²X is O or NH;

b) if n is 0, A is CHCH₂C(O)R²And X is O, then R²Is not provided withAnd is

c) If A is C (O) or CH₂X is NR⁶And R is⁶Is not provided with

wherein

n is 0, 1,2, 3 or 4;

R²is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl; and is

wherein

n is 0, 1,2, 3 or 4;

x is O, NH or NR⁶；

A is C (O), CH₂Or CH-CR³R⁴-C(O)R²；

R²Is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl;

R⁶Is optionally substituted phenyl;

provided that

d) If A is CH-CR³R⁴-C(O)R²X is O or NH;

e) if n is 0, A is CHCH₂C(O)R²And X is O, then R²Is not provided withAnd is

If A is C (O) or CH₂X is NR⁶And R is⁶Is not provided with

wherein

n is 0, 1,2, 3 or 4;

R²is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl; and is

In some embodiments of the compounds of formula III or IIIa described above or below, R³And R⁴Are all hydrogen. In thatIn some embodiments of the compounds of formula III or IIIa described above or below, R³Is optionally substituted alkyl and R⁴Is hydrogen. In some embodiments of the compounds of formula III or IIIa described above or below, R³And R⁴Independently an optionally substituted alkyl group.

In some embodiments of the compounds of formula III or IIIa described above or below, R²Is heteroaryl. In certain embodiments, R²Is optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyridazinyl or optionally substituted pyrazinyl. In certain embodiments, R²Is a 5-membered heteroaryl group. In certain embodiments, the 5-membered heteroaryl is thiophene, benzofuran, pyrrole, thiazole, imidazole, oxazole, pyrazole, or triazole. In certain embodiments, R²Is bicyclic heteroaryl. In certain embodiments, the bicyclic heteroaryl is benzimidazole, benzothiazole, benzoxazole, indazole, quinoline, or naphthyridine.

In some embodiments of the compounds of formula III or IIIa described above or below, R²Is phenyl. In certain embodiments, R²The phenyl group of (a) is disubstituted. In certain embodiments, R²The phenyl group of (a) is monosubstituted. In certain embodiments, R²The substitution on the phenyl group of (a) is independently selected from optionally substituted alkyl, optionally substituted alkoxy, optionally substituted aryloxy, halo, CN, CO₂H. Amino, monoalkylamine, dialkylamine, monoarylamine, alkylarylamine, cycloalkyl, hydroxy, C (O) - (optionally substituted alkyl), C (O) NH₂C (O) NH- (optionally substituted alkyl), alkyl sulfide, alkyl sulfoxide, alkyl sulfone, C (O) - (optionally substituted aryl), C (O) NH- (optionally substituted aryl), aryl sulfide, aryl sulfoxide or aryl sulfone. In certain embodiments, the substitutions on the phenyl groups are independently selected from F, Cl, CO₂H、CN、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂And (5) OH. In some implementationsIn the scheme, R²Is disubstituted by CN and is selected from F, Cl, CO₂H、OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²By disubstituted radicals of CO on the phenyl radical of₂H and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CH₂CH₂CH₂The radical composition of OH. In certain embodiments, R²Disubstituted on the phenyl radical of (A) by CH₂CH₂CH₂OH and is selected from F, Cl, CN, OCH₃、C(O)CH₃、CF₃、CH₃、CH₂OH、CH₂CH₂OH and CO₂The radical composition of H.

In some embodiments of the compounds of formula III or IIIa described above or below, R²Is naphthyl.

In some embodiments described above or below for compounds of formula I, the compound is selected from:

In some embodiments described above or below for compounds of formula Ia, the compound is selected from:

In some embodiments described above or below for compounds of formula Ib, the compound is selected from:

In some embodiments described above or below for compounds of formula Ic, the compound is selected from:

In some embodiments described above or below for compounds of formula II, the compound is selected from:

In some embodiments described above or below for a compound of formula IIa, the compound is selected from:

In some embodiments described above or below for compounds of formula IIb, the compound is selected from:

In some embodiments described above or below for compounds of formula IIc, the compound is selected from:

In some embodiments described above or below for compounds of formula III, the compound is selected from:

in some embodiments, the mammal does not have arthritis or joint damage, but is at increased risk of having arthritis or joint damage.

It is contemplated that the compounds, compositions and methods of the present invention may be used to ameliorate any type of arthritis or joint injury. It is further contemplated that the compounds, compositions, and methods of the invention can be used to ameliorate various cartilage disorders. In some embodiments, the compounds and compositions of the present invention are administered to prevent arthritis or joint damage, for example, in the presence of a genetic or family history or prior or during joint surgery for arthritis or joint damage, or in other situations where the risk of arthritis or joint damage is increased. Exemplary conditions or disorders to be treated or prevented with the compounds, compositions and methods of the invention include, but are not limited to, systemic rheumatoid arthritis, juvenile chronic arthritis, osteoarthritis, degenerative disc disease, spondyloarthropathies and systemic sclerosis (scleroderma). In some embodiments of the invention, the compounds, compositions, and methods of the invention may be used to treat osteoarthritis. In some embodiments, the arthritis can be osteoarthritis, traumatic arthritis, degenerative disc disease, dupuytren's disease, or tendon disease.

In some embodiments, the compounds, compositions and methods of the present invention provide a method for stimulating chondrocyte proliferation and cartilage production in cartilage tissue that has been damaged as a result of traumatic injury or cartilage disease. Traumatic injuries may include, but are not limited to, blunt injuries to joints or injuries to ligaments such as anterior cruciate ligaments, medial collateral ligament tears, or meniscal tears. Examples of tissues that exhibit articulating surfaces and are therefore particularly sensitive to treatment include, but are not limited to, the spine, shoulders, elbows, wrists, finger joints, hips, knees, ankles, and foot joints. Examples of diseases that may benefit from treatment include osteoarthritis, rheumatoid arthritis, other autoimmune diseases, or isolated osteochondritis. In addition, cartilage malformations are common in human dwarfism, suggesting that the compounds, compositions, and methods would be useful in these patients.

It is contemplated that the compounds, compositions and methods of the invention can be used to treat mammals. As used herein, "mammal" refers to any mammal classified as a mammal, including humans, domesticated animals and farm animals, as well as zoo, sports, or pet animals, such as cattle (e.g., cows), horses, dogs, sheep, pigs, rabbits, goats, cats, and the like. In some embodiments, the mammal may be a human, dog, cat, or horse. In some embodiments of the invention, the mammal is a human. In some embodiments, the mammal is a dog, cat, or horse. In some embodiments, the mammal is a cow, sheep, pig, goat, or rabbit. In some embodiments, the mammal is a domesticated animal or a farm animal. In further embodiments, the domesticated animal or livestock is a dog, cat, or horse. In some embodiments, the mammal is a companion animal. As used herein, "companion animal" refers to dogs, cats, rodents, and rabbits. In some embodiments, the mammal is a companion animal or livestock. In some embodiments, the mammal is a domestic animal.

The compounds of the invention may also be used to induce the differentiation of Mesenchymal Stem Cells (MSCs) into chondrocytes. In some embodiments, the present invention provides a method of inducing differentiation of a mesenchymal stem cell into a chondrocyte, the method comprising contacting a mesenchymal stem cell with a sufficient amount of a compound of the present invention, thereby inducing differentiation of the stem cell into a chondrocyte.

MSCs are pluripotent stem cells that are capable of differentiating into several different types of cells, including but not limited to osteoblasts, chondrocytes, and adipocytes. Differentiation is the process of forming specialized cell types from less specialized cell types, such as chondrocytes from MSCs. In some embodiments, the method is performed in vitro. In some embodiments, the method is performed in vivo in a mammal, and the stem cell is present in the mammal. In certain embodiments, the mammal is a human, dog, cat, or horse. In certain embodiments, the mammal is a human. In certain embodiments, the mammal is a dog, cat, or horse.

Inducing differentiation of MSCs into chondrocytes can be accomplished using any suitable amount of a compound of the invention. In some embodiments, the compounds of the present invention may be present in an amount of from about 0.1mg to about 10000mg, such as from 1.0mg to 1000mg, such as from 10mg to 500mg, depending on the particular application and potency of the active ingredient. In some embodiments, the compounds of the present invention may be present in intra-articular injection solutions for the knee at concentrations ranging from 0.1 μ M to 100 μ M.

Assays for identifying compounds

The compounds of the invention are identified using a variety of assays. Initial screening identified compounds that stimulated human mesenchymal stem cells (hmscs) to develop chondrocyte nodules. Additional experiments were performed to determine the toxicity and specificity of chondrocyte differentiation.

Chemical combinationArticle (A)

Described herein are compounds that induce the differentiation of mesenchymal stem cells into chondrocytes. In some embodiments, the compounds described herein ameliorate arthritis or joint damage in a mammal. In some embodiments, the compounds described herein treat arthritis or joint injury in a mammal.

In one aspect, provided herein is a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula Ia, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula Ib, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula Ic, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula II, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula IIa, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula IIb, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula IIc, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

In another aspect, provided herein is a compound of formula III, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof.

preparation of the Compounds

Described herein are compounds for inducing differentiation of mesenchymal stem cells into chondrocytes and for ameliorating arthritis or joint injury in a mammal and methods of making these compounds. Also described herein are pharmaceutically acceptable salts, pharmaceutically acceptable solvates, pharmaceutically active metabolites, and pharmaceutically acceptable prodrugs of these compounds. Also provided are pharmaceutical compositions comprising at least one such compound or a pharmaceutically acceptable salt, pharmaceutically acceptable solvate, pharmaceutically active metabolite, or pharmaceutically acceptable prodrug of the compound, and a pharmaceutically acceptable excipient.

The compounds described herein can be synthesized using standard synthetic reactions known to those skilled in the art or using methods known in the art. The compounds may be provided using reactions in a linear order, or the reactions may be used to synthesize fragments, which are then ligated by methods known in the art.

The starting materials for synthesizing the compounds described herein may be synthesized or may be obtained from commercial sources such as, but not limited to, Aldrich Chemical co. (Milwaukee, Wisconsin), Bachem (Torrance, California), or Sigma Chemical co. (st. The compounds described herein and other related compounds having different substituents can be synthesized using techniques and materials known to those skilled in the art, for example using techniques and materials described in, for example: march, ADVANCED ORGANIC CHEMISTRY, 4 th edition, (Wiley 1992); carey and Sundberg, ADVANCED ORGANIC CHEMISTRY, 4 th edition, Vols.A and B (Plenum 2000, 2001); green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, 3 rd edition, (Wiley 1999); fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-17(John Wiley and Sons, 1991); rodd's Chemistry of Carbon Compounds, Volumes 1-5and supplements (Elsevier Science Publishers, 1989); organic Reactions, Volumes 1-40(John Wileyandsons, 1991); and Larock's Comprehensive Organic Transformations (VCHPublishers Inc., 1989). (all of which are incorporated herein by reference in their entirety). Other methods for synthesizing the compounds described herein can be found in International patent publication No. WO 01/01982901, Arnold et al, Bioorganic & medicinal chemistry Letters 10(2000) 2167-2170; burcha et al, Bioorganic & Medicinal chemistry letters 12(2002) 1687-1690. General methods for preparing the compounds disclosed herein can be derived from reactions known in the art, and such reactions can be modified, as recognized by those skilled in the art, by the use of appropriate reagents and conditions in order to introduce the various moieties found in the general formulae provided herein.

If desired, the reaction product may be isolated and purified using conventional techniques including, but not limited to, filtration, distillation, crystallization, and chromatography. Such materials may be characterized using conventional means including physical constants and spectroscopic data.

The compounds described herein may be prepared as a single isomer or as a mixture of isomers.

Other forms of the compounds disclosed herein

Isomers

In some embodiments, the compounds described herein exist in geometric isomeric forms. In some embodiments, the compounds described herein have one or more double bonds. The compounds provided herein include all cis, trans, ipsilateral, contralateral, E (E) and Z (Z) isomers, as well as their corresponding mixtures. In some cases, the compounds exist in tautomeric forms. The compounds described herein include all possible tautomers within the general formulae described herein. In some cases, the compounds described herein have one or more chiral centers and each center is present in the R configuration or the S configuration. The compounds described herein include all diastereomeric, enantiomeric and epimeric forms and the corresponding mixtures thereof. In other embodiments of the compounds and methods provided herein, mixtures of enantiomers and/or diastereomers resulting from a single preparation step, combination, or interconversion can be used for the applications described herein. In some embodiments, the compounds described herein are prepared as their individual stereoisomers by reacting a racemic mixture of the compounds with an optical resolution agent to form a pair of diastereomeric compounds, separating the diastereomers and recovering the optically pure enantiomers. In some embodiments, isolatable complexes (e.g., crystalline diastereomeric salts) are preferred. In some embodiments, diastereomers have different physical properties (e.g., melting points, boiling points, solubilities, reactivities, etc.), and separation is performed using these differences. In some embodiments, the diastereomers are separated by chiral chromatography or, preferably, by separation/resolution techniques based on solubility differences. In some embodiments, the optically pure enantiomer is then recovered along with the resolving agent by any practical means that does not cause racemization.

Labelled compounds

In some embodiments, the compounds described herein are present in their isotopically labeled form. In some embodiments, the methods disclosed herein include methods of treating diseases by administering such isotopically labeled compounds. In some embodiments, the methods disclosed herein include methods of treating diseases by administering such isotopically labeled compounds in the form of pharmaceutical compositions. Thus, in some embodiments, the compounds disclosed herein include isotopically labeled compounds, which are identical to those recited herein, except for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, respectively²H、³H、¹³C、¹⁴C、¹⁵N、¹⁸O、¹⁷O、³¹P、³²P、³⁵S、¹⁸F and³⁶and (4) Cl. Compounds described herein, and metabolites, pharmaceutically acceptable salts, esters, prodrugs, solvates, hydrates, or derivatives thereof, containing the aforementioned isotopes and/or other isotopes of other atoms are within the scope of the present invention. Certain isotopically-labelled compounds, e.g. in which a radioactive isotope such as³H and¹⁴c, useful in drug and/or stromal tissue distribution assays. Tritium labeling (i.e.³H) And carbon-14 (i.e.¹⁴C) Isotopes are particularly preferred for their ease of preparation and detectability. In addition, heavy isotopes such as deuterium (i.e. deuterium) are used²H) Substitution produces certain therapeutic advantages resulting from higher metabolic stability, such as an increase in vivo half-life or a reduction in dosage requirements. In some embodiments, the isotopically-labeled compound, pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof is prepared by any suitable method.

In some embodiments, the compounds described herein are labeled by other means, including but not limited to the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

Pharmaceutically acceptable salts

In some embodiments, the compounds described herein are present in the form of a pharmaceutically acceptable salt thereof. In some embodiments, the methods disclosed herein include methods of treating a disease by administering such pharmaceutically acceptable salts. In some embodiments, the methods disclosed herein include methods of treating diseases by administering such pharmaceutically acceptable salts in the form of pharmaceutical compositions.

In some embodiments, the compounds described herein have acidic or basic groups and thus react with any of a variety of inorganic or organic bases and inorganic and organic acids to form pharmaceutically acceptable salts. In some embodiments, these salts are prepared in situ during the final isolation and purification of the compounds of the present invention, or by reacting the purified compound in free form with a suitable acid or base, respectively, and isolating the salt thus formed.

Examples of pharmaceutically acceptable salts include those salts prepared by reacting a compound described herein with an inorganic acid, organic acid, or inorganic base, such salts including acetate, acrylate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bromide, butyrate, butyne-1, 4-dioate, camphorate, camphorsulfonate, hexanoate, octanoate, chlorobenzoate, chloride, citrate, cyclopentanepropionate, decanoate, digluconate, dihydrogenphosphate, dinitrobenzoate, dodecylsulfate, ethanesulfonate, formate, fumerate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hexyne-1, 6-dioate, hydroxybenzoate, dihydroxybenzoate, ethylsulfonate, formate, fumarate, dihydroxybenzoate, gamma-hydroxybutyrate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isobutyrate, lactate, maleate, malonate, methanesulfonate, mandelate, metaphosphate, methanesulfonate, methoxybenzoate, methylbenzoate, monohydrogenphosphate, 1-naphthalenesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, pamoate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate, phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate, sulfate, sulfite, succinate, suberate, sebacate, sulfonate, tartrate, di-n-butyl acetate, di-butyl acetate, thiocyanate, tosylate, undecanoate, and xylenesulfonate.

Further, the compounds described herein can be prepared as pharmaceutically acceptable salts formed by reacting the free base form of the compound with pharmaceutically acceptable inorganic or organic acids, including, but not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, and the like; and organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, Q-toluenesulfonic acid, tartaric acid, trifluoroacetic acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, arylsulfonic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo- [2.2.2] oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4' -methylenebis- (3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, Lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid. In some embodiments, other acids, such as oxalic acid, while not per se pharmaceutically acceptable, are used in the preparation of salts useful as intermediates in obtaining the compounds of the present invention and their pharmaceutically acceptable acid addition salts.

In some embodiments, those compounds described herein that contain a free acid group are reacted with: suitable bases, such as pharmaceutically acceptable basesThe metal cation is selected from the group consisting of hydroxide, carbonate, bicarbonate, sulfate, ammonia, or a pharmaceutically acceptable organic primary, secondary, tertiary, or quaternary amine. Representative salts include alkali metal or alkaline earth metal salts such as lithium, sodium, potassium, calcium and magnesium salts, and aluminum salts and the like. Illustrative examples of the base include sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, N⁺(C_1-4Alkyl radical)₄And the like.

Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like. It is to be understood that the compounds described herein also include the quaternization of any basic nitrogen-containing groups they contain. In some embodiments, water-soluble or oil-soluble or dispersible products are obtained by such quaternization.

Solvates

In some embodiments, the compounds described herein are present in the form of solvates. The present invention provides methods of treating diseases by administering such solvates. The invention further provides methods of treating diseases by administering such solvates in the form of a pharmaceutical composition.

Solvates contain stoichiometric or non-stoichiometric amounts of solvent and, in some embodiments, are formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, and the like. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is alcohol. Solvates of the compounds described herein may conveniently be prepared or formed in the processes described herein. By way of example only, hydrates of the compounds described herein may be conveniently prepared by recrystallization from aqueous/organic solvent mixtures using organic solvents including, but not limited to, dioxane, tetrahydrofuran, or methanol. In addition, the compounds provided herein can exist in unsolvated as well as solvated forms. In general, the solvated forms are considered equivalent to unsolvated forms for the compounds and methods provided herein.

Polymorphic substance

In some embodiments, the compounds described herein exist in the form of polymorphs. The present invention provides methods of treating diseases by administering such polymorphs. The invention further provides methods of treating diseases by administering such polymorphs in the form of pharmaceutical compositions.

Accordingly, the compounds described herein include all crystalline forms thereof, referred to as polymorphs. Polymorphs include different crystal packing arrangements of a compound having the same elemental composition. In some cases, polymorphs have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shape, optical and electrical properties, stability, and solubility. In some cases, factors such as recrystallization solvent, crystallization rate, and storage temperature make a single crystal form dominant.

Prodrugs

In some embodiments, the compounds described herein are present in the form of a prodrug. The present invention provides methods of treating diseases by administering such prodrugs. The invention further provides methods of administering such prodrugs in the form of pharmaceutical compositions to treat diseases.

Prodrugs are generally prodrugs that, upon administration to an individual and subsequent absorption, are converted to an active or more active substance via a process such as conversion by metabolic pathways. Some prodrugs have chemical groups on the prodrug that render it less active and/or impart solubility or some other property to the drug. Once the chemical group is cleaved and/or modified from the prodrug, the active drug is produced. Prodrugs are often useful because, in some cases, they are easier to administer than the parent drug. For example, they may be bioavailable by oral administration whereas the parent drug is not. In some cases, the prodrug also has improved solubility in pharmaceutical compositions over the parent drug. One example of a prodrug is, but is not limited to, a compound as described herein, which is administered in the form of an ester ("prodrug") to facilitate transport across a cell membrane where water solubility is detrimental to mobility, but which is then metabolically hydrolyzed to a carboxylic acid (active entity) once inside the cell where water solubility is favorable. Another example of a prodrug may be a short peptide (polyamino acid) bonded to an acid group, where the peptide is metabolized to reveal the active moiety. (see, e.g., Bundgaard, "Design and Application of primers", A Textbook of Drug Design and development, Krosgaard-Larsen and Bundgaard eds, 1991, Chapter 5, 113-.

In some embodiments, prodrugs are designed as reversible drug derivatives to be used as modulators to enhance drug delivery to site-specific tissues. Prodrugs have been designed to increase the effective water solubility of therapeutic compounds targeted to regions where water is the predominant solvent so far.

In some embodiments, the prodrug is C of a compound disclosed herein₁-C₆An alkyl ester.

In addition, prodrug derivatives of the compounds described herein can be prepared by methods described herein or known in the art (for more details, see Saulnier et al, Bioorganic and Medicinal Chemistry Letters,1994,4, 1985). By way of example only, suitable prodrugs can be prepared by reacting an underivatized compound with a suitable carbamoylation reagent such as, but not limited to, 1-acyloxyalkyl chloroformate, p-nitrophenylcarbonate, and the like. Prodrug forms of the compounds described herein (where the prodrug is metabolized in vivo to produce a derivative as described herein) are included within the scope of the claims. Indeed, some of the compounds described herein are prodrugs of another derivative or active compound.

In some embodiments, prodrugs include compounds in which an amino acid residue or a polypeptide chain having two or more (e.g., 2, 3, or 4) amino acid residues is covalently attached via an amide or ester bond to a free amino, hydroxyl, or carboxylic acid group of a compound of the invention. Amino acid residues include, but are not limited to, the 20 naturally occurring amino acids, and also include 4-hydroxyproline, hydroxylysine, desmosine (destosine), isodesmosine, 3-methylhistidine, norvaline, β -alanine, γ -aminobutyric acid, citrulline, homocysteine, homoserine, ornithine and methionine sulfone. In other embodiments, prodrugs include compounds wherein a nucleic acid residue or an oligonucleotide having two or more (e.g., 2, 3, or 4) nucleic acid residues is covalently attached to a compound of the invention.

Pharmaceutically acceptable prodrugs of the compounds described herein also include, but are not limited to, esters, carbonates, thiocarbonates, N-acyl derivatives, N-acyloxyalkyl derivatives, quaternized derivatives of tertiary amines, N-Mannich bases, Schiff bases, amino acid conjugates, phosphates, metal salts, and sulfonates. Compounds having a free amino, amido, hydroxyl or carboxyl group may be converted into prodrugs. For example, the free carboxyl groups may be derivatized as amides or alkyl esters. In some cases, groups including, but not limited to, ether, amine, and carboxylic acid functional groups are incorporated into all of these prodrug moieties.

Hydroxy prodrugs include esters such as, but not limited to, acyloxyalkyl (e.g., acyloxymethyl, acyloxyethyl) esters, alkoxycarbonyloxyalkyl esters, alkyl esters, aryl esters, phosphate esters, sulfonate esters, sulfate esters, and disulfide-containing esters; ethers, amides, carbamates, hemisuccinates, dimethylaminoacetates and phosphoryloxymethoxymethoxycarbonyls, as outlined in advanced drug Delivery Reviews 1996,19, 115.

Amine-derived prodrugs include, but are not limited to, the following groups and combinations of groups:

and sulfonamides and phosphoramides.

In some cases, various metabolic reactions are prone to occur at sites on any aromatic ring moiety, and thus the incorporation of suitable substituents on the aromatic ring structure can reduce, minimize or eliminate this metabolic pathway.

Metabolites

In some embodiments, the compounds described herein are susceptible to various metabolic reactions. Thus, in some embodiments, incorporation of suitable substituents into the structure will reduce, minimize or eliminate metabolic pathways. In particular embodiments, suitable substituents that reduce or eliminate the sensitivity of the aromatic ring to metabolic reactions are, by way of example only, halogen or alkyl.

In additional or further embodiments, the compounds described herein are metabolized upon administration to an organism in need thereof to produce a metabolite that is subsequently used to produce a desired effect, including a desired therapeutic effect.

Pharmaceutical composition/formulation

In another aspect, provided herein is a pharmaceutical composition comprising a compound described herein, or a pharmaceutically acceptable salt, polymorph, solvate, prodrug, N-oxide, or isomer thereof, and a pharmaceutically acceptable excipient.

In some embodiments, the compounds described herein are formulated as pharmaceutical compositions. The pharmaceutical compositions are formulated in conventional manner using one or more pharmaceutically acceptable inactive ingredients that facilitate processing of the active compounds into preparations that can be used pharmaceutically. The appropriate formulation will depend on the route of administration chosen. A summary of The pharmaceutical compositions described herein can be found, for example, in Remington: The Science and Practice of Pharmacy, nineteenth edition (Easton, Pa.: Mack Publishing Company, 1995); hoover, John e., Remington's pharmaceutical Sciences, Mack Publishing co., Easton, Pennsylvania 1975; liberman, h.a. and Lachman, l. eds, Pharmaceutical document Forms, Marcel Decker, new york, n.y., 1980; and Pharmaceutical document Forms and Drug Delivery Systems, seventh edition (Lippincott Williams & Wilkins 1999), which publications are incorporated herein by reference.

Provided herein are pharmaceutical compositions comprising a compound described herein and at least one pharmaceutically acceptable inactive ingredient. In some embodiments, the compounds described herein are administered as a pharmaceutical composition in which the compounds described herein are mixed with other active ingredients, such as in a combination therapy. In other embodiments, the pharmaceutical composition comprises other medical or pharmaceutical preparations, carriers, adjuvants, preservatives, stabilizers, wetting or emulsifying agents, dissolution promoters (solubility promoters), salts for regulating osmotic pressure and/or buffers. In still other embodiments, the pharmaceutical composition comprises other therapeutically valuable substances.

As used herein, a pharmaceutical composition refers to a mixture of a compound described herein with other chemical components (i.e., pharmaceutically acceptable inactive ingredients) such as carriers, excipients, binders, fillers, suspending agents, flavoring agents, sweeteners, disintegrants, dispersants, surfactants, lubricants, colorants, diluents, solubilizers, wetting agents, plasticizers, stabilizers, permeation enhancers, wetting agents, antifoaming agents, antioxidants, preservatives, or one or more combinations thereof. The pharmaceutical composition facilitates administration of the compound to an organism. In practicing the methods of treatment or uses provided herein, a therapeutically effective amount of a compound described herein is administered in the form of a pharmaceutical composition to the mammal to be treated suffering from the disease, disorder, or condition. In some embodiments, the mammal is a human, dog, cat, or horse. In some embodiments, the mammal is a human. In some embodiments, the mammal is a dog, cat, or horse. The therapeutically effective amount may vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds may be used alone or as components of a mixture in combination with one or more therapeutic agents.

The pharmaceutical formulations described herein are administered to a subject by a suitable route of administration, including, but not limited to, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular, intraarticular), intranasal, buccal, topical, rectal, or transdermal routes of administration. Pharmaceutical formulations described herein include, but are not limited to: aqueous liquid dispersions, liquids, gels, syrups, elixirs, slurries, suspensions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid oral dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, powders, lozenges, effervescent formulations, lyophilized formulations, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations and mixed immediate and controlled release formulations.

Pharmaceutical compositions containing a compound described herein are prepared in conventional manner, such as, for example only, by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or compressing methods.

The pharmaceutical compositions will comprise as active ingredient at least one compound described herein, in the form of a free acid or a free base or in the form of a pharmaceutically acceptable salt. In addition, the methods and pharmaceutical compositions described herein include the use of N-oxides (if appropriate), crystalline forms, amorphous phases, and active metabolites of these compounds that have the same type of activity. In some embodiments, the compounds described herein exist in unsolvated forms or exist in solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. Solvated forms of the compounds provided herein are also considered disclosed herein.

The pharmaceutical preparation for oral use is obtained by the following method: one or more solid excipients are mixed with one or more compounds as described herein, the resulting mixture is optionally ground, and the mixture of granules is processed, after addition of suitable auxiliaries (if desired), to give tablets or dragee cores. Suitable excipients include, for example, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations, for example, corn starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, microcrystalline cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose; or other excipients, such as: polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. If desired, disintegrating agents are added, such as croscarmellose sodium, polyvinylpyrrolidone, agar or alginic acid or a salt thereof such as sodium alginate. In some embodiments, dyes or pigments are added to the tablets or lozenge coatings for identifying or characterizing different combinations of active compound doses.

Pharmaceutical formulations for oral administration include push-fit (push-fit) capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Push-fit capsules contain the active ingredients in admixture with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds are dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In some embodiments, a stabilizer is added.

In certain embodiments, delivery systems for pharmaceutical compounds may be employed, for example, liposomes and emulsions. In certain embodiments, the compositions provided herein can further include a mucoadhesive polymer selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly (methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid/butyl acrylate copolymer, sodium alginate, and dextran.

Combination therapy

The compounds and compositions of the present invention may be used in combination with other components suitable for ameliorating arthritis or joint damage. In some embodiments, the composition may further comprise an additional compound that is therapeutically effective for treating arthritis or joint damage and/or symptoms associated with arthritis or joint damage in a mammal. In some embodiments, the composition may further comprise a non-steroidal anti-inflammatory drug (NSAID), an analgesic, a glucocorticoid, angiopoietin-like 3 protein (ANGPTL3) or chondrogenic variant thereof, oral salmon calcitonin, SD-6010(iNOS inhibitor), vitamin D3 (cholecalciferol), collagen hydrolysate, FGF18, BMP7, Avocado Soy Unsaponifiable (ASU), or hyaluronic acid. ANGPTL3 is described in more detail in WO 2011/008773 (incorporated herein in its entirety). In some embodiments, the composition comprises an agent with anti-inflammatory activity. In some embodiments, the composition comprises an apoptosis modulator. In certain embodiments, the apoptosis modulator is a caspase inhibitor. One non-limiting example of an apoptosis/caspase inhibitor is enrichloron. In some embodiments, the composition comprises an iNOS inhibitor. One non-limiting example of an iNOS inhibitor is SD-6010.

NSAIDs include, but are not limited to, aspirin, diflunisal, salsalate, ibuprofen, dexibuprofen, naproxen, fenoprofen, ketoprofen, dexketoprofen, flurbiprofen, oxaprozin, loxoprofen, indomethacin, tolmetin, sulindac, etodolac, ketorolac, nabumetone, diclofenac, piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, isoxicam, mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamic acid, celecoxib, parecoxib, etoricoxib, lumiracoxib, and felicoxib.

Analgesics include, but are not limited to, acetaminophen and opioids (narcotics). Opioids include, but are not limited to, dextropropoxyphene, codeine, tramadol, tapentadol, anileridine, alfacidine, meperidine, hydrocodone, morphine, oxycodone, methadone, diacetylmorphine, hydromorphone, oxymorphone, levorphanol, 7-hydroxycodesine, buprenorphine, fentanyl, sufentanil, bromodol, etorphine, dihydroetorphine, and carfentanil.

Glucocorticoids include, but are not limited to, hydrocortisone, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, or fludrocortisone.

The compounds described herein may be used in combination with one or more compounds that are therapeutically effective for treating arthritis or joint damage and/or symptoms associated with arthritis or joint damage. Such additional compounds may be administered by conventional routes and in conventional amounts, simultaneously or sequentially with the compounds disclosed herein. When the compounds disclosed herein are used contemporaneously with one or more of such additional compounds, a pharmaceutical composition in unit dosage form containing such other drugs and a compound of the present invention is preferred. However, combination therapy may also include therapies in which a compound disclosed herein and one or more additional compounds are administered according to different overlapping schedules. It is also contemplated that when used in combination with one or more additional compounds, the compounds may be used in lower doses than when each is used alone.

The above combinations include combinations of the compounds disclosed herein not only with one compound that is therapeutically effective for treating arthritis or joint damage and/or symptoms associated with arthritis or joint damage, but also with two or more such compounds. Likewise, the compounds disclosed herein, by themselves or in combination with compounds that are therapeutically effective for treating arthritis or joint damage and/or symptoms associated with arthritis or joint damage, may be used in combination with other drugs used in the prevention, treatment, control, or amelioration of osteoarthritis or joint damage or conditions associated with osteoarthritis or joint damage. Such other agents may be administered by the usual routes and in the usual amounts, simultaneously or sequentially with the compounds disclosed herein. When the compounds disclosed herein are used contemporaneously with one or more other drugs, pharmaceutical compositions containing such other drugs in addition to the compounds of the present invention are preferred. Accordingly, the pharmaceutical compositions of the present invention also include those that contain one or more other active ingredients in addition to the compounds disclosed herein. The weight ratio of the compound disclosed herein to the second active ingredient can vary and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used.

Administration of pharmaceutical compositions

Suitable routes of administration include, but are not limited to, oral, intravenous, intra-articular, rectal, spray, parenteral, ocular, pulmonary, transmucosal, transdermal, vaginal, otic, nasal, and topical administration. Further, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injections, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, intraarticular, and intranasal injections.

In some embodiments, the compounds disclosed herein and compositions thereof are administered in any suitable manner. The mode of administration may be selected based on, for example, whether local or systemic treatment is desired and the area to be treated. For example, the compositions can be administered orally, parenterally (e.g., intravenous, subcutaneous, intraperitoneal, intra-articular, or intramuscular injection), by inhalation, extracorporeally, topically (including transdermal, ocular, vaginal, rectal, intranasal), and the like. In some embodiments, the composition may be administered by a microneedle. In some embodiments, the composition may be administered via a microneedle array in the form of a patch capable of intradermal drug delivery. In some embodiments, the composition can be administered by transdermal microneedle patch delivery.

Parenteral administration of the composition, if used, is generally characterized by injection. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for dissolution or suspension in liquid prior to injection, or as emulsions. A recently revised approach for parenteral administration includes the use of slow or delayed release systems to maintain a constant dose.

Examples

List of abbreviations

As used above, and throughout the specification of the present invention, the following abbreviations, unless otherwise indicated, shall be understood to have the following meanings:

ACN acetonitrile

Bn phenyl

BOC or Boc carbamic acid tert-butyl ester

BOP benzotriazol-1-yl-oxytis (dimethylamino) phosphonium

t-Bu tert-butyl

Cbz Carboxylic acid benzyl ester

Cy cyclohexyl group

DBU 1, 8-diazabicyclo [5.4.0] undec-7-ene

DCC dicyclohexylcarbodiimide

DCM dichloromethane (CH2CI2)

DIC 1, 3-diisopropylcarbodiimide

DEAD azodicarboxylic acid diethyl ester

DIAD diisopropyl azodicarboxylate

DIPEA diisopropylethylamine

DMAP 4- (N, N-dimethylamino) pyridine

DMP reagent Dess-Martin oxidant (Dess-Martin Periodinane reagent)

DMF dimethyl formamide

DMA N, N-dimethylacetamide

DME 1, 2-dimethoxy-ethane

DMSO dimethyl sulfoxide

Dppf 1,1' -bis (diphenylphosphino) ferrocene

EDCI 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide HCl

eq equivalent weight

Et Ethyl group

Et2O diethyl ether

EtOH ethanol

EtOAc ethyl acetate

HOAt 1-hydroxy-7-azabenzotriazole

HOBT 1-hydroxybenzotriazole

HOSu N-hydroxysuccinamide

HPLC high performance liquid chromatography

LAH anhydride lithium aluminium

Me methyl group

MeI methyl iodide

MeOH methanol

MOMCl methoxymethyl chloride

MOM methoxymethyl group

MS mass spectrometry

NMP N-methyl-pyrrolidin-2-one

NMR nuclear magnetic resonance

PyBOP benzotriazol-1-yl-oxytri-pyrrolidinyl-phosphonium hexafluorophosphate

SPHOS 2-dicyclohexylphosphino-2 ',6' -dimethoxybiphenyl

TBD 1,5, 7-triazabicyclo [4.4.0] -dec-5-ene

RP-HPLC reversed-phase high-pressure liquid chromatography

TBS tert-butyldimethylsilyl radical

TBSCl tert-butyldimethylsilyl chloride

TBTU O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethylonium

TEOC 2-trimethylsilylethyl carbamate

TFA trifluoroacetic acid

Tf2O Trifluoromethanesulfonic anhydride

TMG 1,1,3, 3-tetramethylguanidine

THF tetrahydrofuran

THP tetrahydropyrans

TLC thin layer chromatography

XPHOS 2-dicyclohexylphosphino-2 ', 4', 6' -triisopropylbiphenyl

General examples for the preparation of the compounds of the invention

Starting materials and intermediates for the compounds of the invention can be prepared by applying or modifying the methods described below, their obvious chemical equivalents, or, for example, as described in the following documents: the Science of Synthesis, Vol.1-8, E.M. Carreira et al, eds. Thieme publishers (2001-. Details of reagents and reaction options can also be obtained by conducting structure and reaction searches using commercial computer search engines such as Scfiner (www.cas.org) or Reaxys (www.reaxys.com).

Synthetic examples

The following preparations of the compounds and intermediates disclosed herein are given to enable those skilled in the art to more clearly understand and practice the present invention. They should not be considered as limiting the scope of the invention, but merely as being exemplary and representative thereof.

Synthetic scheme a: experimental samples for Compound 1

To an EtOAc solution of phthalic anhydride (1.0eq) was added 3-amino-4-chlorobenzonitrile (1eq) followed by stirring at 20-30 ℃ for 1-8 h. TLC indicated that the starting material had disappeared. The reaction mixture was filtered and the solid was purified by recrystallization from EtOAc to give compound 1(12 mg). End product 1 by¹H NMR and LCMS. LCMS: found 301.0[ M + H]。¹H NMR(400MHz,MeOD-d₄):8.35(s,1H),8.09(d,J＝7.7Hz,1H),7.71(dd,J＝8.0,4.1Hz,1H),7.67(d,J＝8.4Hz,1H),7.61(t,J＝7.4Hz.2H),7.57(dd,J＝8.4,2.0Hz,1H)。

The selected compounds in table 1 were obtained using similar conditions to the reaction scheme given above, wherein 3-amino-4-chlorobenzonitrile was replaced with the appropriate aniline or amine. The reaction yield based on the isolated product ranged from 20% to 80%.

Synthesis scheme B-1: experimental sample for Compound 15

To a solution of 4-aminobenzenesulfonamide (100mg, 0.58mmol) in AcOH (20mL) was added phthalic anhydride (82mg, 0.55 mmol). The mixture was stirred at 130 ℃ for 3 h. The mixture is treated with H₂Diluted O (30mL) and stirred for 2 h. After filtration, Compound 15(44mg, yield: 26%) was obtained as a white solid. End product 15 by¹H NMR and LCMS. LCMS: found 303.0[ M + H ]]。¹H NMR(400MHz,DMSO-d₆):7.10-7.36(m,6H),6.56-6.70(m,2H),6.62(br s,2H)。

Selected compounds in table 1 were obtained using similar conditions to the reaction scheme given above, with the replacement of 4-aminobenzenesulfonamide with the appropriate aniline. In the case of compound 33, the reaction time was 3H and the reaction mixture was washed with H₂O diluted and stirred for 12h for crystallization.

Synthesis scheme B-2: experiment for Compound 26Sample (I)

To a solution of compound 15(276mg, 0.913mmol) in AcOH (5mL) was added Zn (596.91mg, 9.13mmol) and DMF (0.1 mL). The mixture was stirred at 130 ℃ for 3h, then cooled to room temperature and concentrated to give the crude product as a colourless oil. The residue was purified by preparative HPLC (0.1% TFA as additive) and most of the CH was removed by evaporation under reduced pressure₃CN, and the remaining solvent was removed by lyophilization to give compound 26 as a white solid (80mg, yield: 30%). LCMS: found 289.1[ M + H ]]。

Compound 27 (table 1) was obtained using similar conditions as the reaction scheme given above. In the preparation of compound 27, DMF was not used.

The starting material for product 27 was prepared by the following procedure.

Preparation of Compound 27-2

To a solution of compound 27-1(3.8g, 17mmol) in DCM (50ml) was added MeNH in alcohol₂(5.3g, 51 mmol). The mixture was stirred at room temperature for 2 h. The mixture was diluted with DCM (30ml) and H₂O (30ml) wash. The organic layer was passed over anhydrous Na₂SO₄Drying, filtration and concentration gave compound 27-2(3.3g, yield: 90%).

Preparation of Compound 27-3

To CH of Compound 27-2(3.3g, 15.3mol) at room temperature₃To the OH (50mL) solution was added Pd/C (0.16 g). The mixture is in H₂(30psi) at room temperature for 12 h. The mixture was filtered and the solvent was removed to give compound 27-3(2.3g, yield: 80%) as a gray solid.

Preparation of starting Material for Compound 27

To a solution of 27-3(0.5g, 2.68mmol) in AcOH (40mL) was added 27-4(0.433g, 2.92 mmol). The reaction mixture was stirred at 130 ℃ for 3 h. The reaction mixture was cooled and the solvent was removed to give the crude product, which was used directly in the next step.

Synthetic scheme B-3:experimental sample for Compound 177

To the starting material of compound 27 (1.0g, 3.16mmol) in CH at 0 deg.C₃NaBH was added to a solution in OH (20mL)₄(239mg, 6.32 mmol). The mixture was stirred at 0 ℃ for 15min, after which the solvent was removed under reduced pressure. To the crude residue was added water (20mL) and saturated NH₄Aqueous Cl (20 mL). The mixture was stirred at room temperature for 30 min. Filtration afforded compound 177 as an off-white solid (840mg, yield: 83%).

Synthetic scheme C: general procedure for Compound C

To a solution of ketone sm-1(1.0eq) in dioxane (V/M10: 1) was added carboxybenzaldehyde sm-2(1.2eq) followed by Sc (OTf)₄. The mixture was heated to reflux for 12 h. After the mixture is cooled to the room temperature,the mixture was concentrated and purified by preparative HPLC to give compound C.

Selected compounds in table 1 were obtained using synthesis scheme C. The reaction yield based on the isolated product ranged from 5% to 50%.

Synthetic scheme D: general procedure for Compound D

To a solution of ketone sm-1(1.0eq) in dioxane (V/M15: 1) was added carboxybenzaldehyde sm-2(1.2eq) followed by Sc (OTf)₄(2 eq). The mixture was heated to reflux for 12 h. Addition of NH₄OAc (5eq) and the reaction mixture was heated to reflux for another 12 h. After cooling to room temperature, the mixture was concentrated and purified by preparative HPLC to give compound D.

Selected compounds in table 1 were obtained using synthesis scheme D. The reaction yield based on the isolated product ranged from 3% to 20%.

Synthetic scheme E: general procedure for Compound E

To a solution of 2-aminobenzoic acid sm-4(1eq) in 2M NaHCO₃To the solution in (V ═ 10eq) was added sulfonyl chloride sm-3(1.0eq) and the mixture was stirred at room temperature for 2 h. TLC indicated that the starting material had disappeared. The mixture was extracted with EtOAc. The organic layer was passed over anhydrous Na₂SO₄Dried, concentrated and purified by preparative HPLC to give pure product E.

Compound 53 (table 1) was obtained using synthesis scheme E. The reaction yield based on the isolated product ranged from 60% to 80%.

Synthetic scheme F: for the compound FGeneral procedure of

EDCI (1.5eq) and HOBT (1.5eq) were added to a solution of sm-6(1eq) benzoic acid in DMF (10eq) at 0 ℃ followed by stirring at room temperature for about 2 h. To the reaction mixture was added amine sm-5(1.5eq) and stirred at room temperature for an additional 12 h. Water was added and the mixture was extracted with EtOAc over anhydrous Na₂SO₄Dried and concentrated. The residue was purified by column chromatography to give product F.

Selected compounds in table 1 were obtained using synthesis scheme F. The reaction yield based on the isolated product ranged from 20% to 40%.

Synthetic scheme G: experimental sample for Compound 56

To a solution of 2-amino-N, N-dimethylbenzenesulfonamide (1.0eq) and TEA (1.5eq) in DCM was added 4-cyanobenzoyl chloride (1.0 eq). The reaction mixture was stirred at 29 ℃ for about 2 h. TLC indicated that the starting material had disappeared. The reaction mixture was washed with saturated NaHCO₃Quench the aqueous solution, extract with DCM, over anhydrous Na₂SO₄Dried and concentrated. The residue was purified by preparative HPLC to give compound 56 (yield: 70%). LCMS: found 330.1[ M + H ]]。

Compound 62 (table 1) was obtained using similar conditions as the reaction scheme given above. The reaction yield based on the isolated product ranged from 70% to 80%.

The aniline starting material I necessary for compounds 56 and 62 was prepared by the following procedure.

Preparation of intermediate sm-8

A solution of 2-nitrobenzene-1-sulfonyl chloride (1.0eq) and amine sm-7(1.0eq) in DCM was stirred at 30 ℃ for about 2 h. TLC indicated that the starting material had disappeared. The reaction mixture was washed with brine and over anhydrous Na₂SO₄Dried and concentrated to give intermediate sm-8 (yield: 72-91%).

Preparation of Aniline starting Material I

To a solution of intermediate sm-8(1.0eq) in MeOH was added Pd/C. Reaction mixture at 50psi H₂The mixture was stirred at 50 ℃ for 7 hours. TLC indicated that the starting material had disappeared. The mixture was filtered, and the filtrate was concentrated to give compound I (yield: 89-91%).

Synthetic scheme H: general procedure for Compound H

To a solution of the amide sm-10(1eq) in THF (V ═ 10eq) at 0 ℃ was added LiHMDS (1eq) dropwise. After 30min, sulfonyl chloride sm-9(1.0eq) was added and the mixture was stirred at room temperature for 2 h. TLC indicated that the starting material had disappeared. The mixture was extracted with EtOAc. The organic layer was passed over anhydrous Na₂SO₄Dried, concentrated, and purified by preparative HPLC to give compound H.

The selected compounds in table 1 were used in synthesis scheme H. The reaction yield based on the isolated product ranged from 50% to 80%.

Synthetic scheme I: general procedure for Compound I

To a stirred mixture of 2-bromo-N- (4-cyanophenyl) benzamide (1.66mmol) and sm-11 borate (3.32mmol) in dioxane (10mL) was added K₃PO₄(1.06g, 4.98 mmol). In N₂Adding Pd₂(dba)₃(45.61mg, 49.81umol) and X-Phos (39.58mg, 83.02 umol). Finally the mixture was heated to 110 ℃ and stirred for 12 h. After filtration, the mixture was concentrated to give intermediate sm-12 as a brown oil. To a solution of sm-12(770mg, 2.16mmol) in THF (30mL) was added LiOH (4.32mL, 4.32mmol) dropwise and stirred for 12 h. The solution was acidified to pH4 at 10 deg.C, extracted with EtOAc (30mL), and extracted with H₂O (50mL) and brine (50mL) were washed, concentrated, and purified by preparative HPLC (0.1% TFA as additive). The solvent was removed by evaporation under reduced pressure and freeze-drying to give compound I as a white solid.

Selected compounds in table 1 were obtained using synthesis scheme I. The reaction yield based on the isolated product ranged from 15% to 30%.

2-bromo-N- (4-cyanophenyl) benzamide was prepared by the following procedure.

2-Bromobenzoic acid (4.4g, 21.89mmol) in SOCl₂The solution (45mL) was heated to 70 ℃ for 2 h. It was evaporated to give 2-bromobenzoyl chloride as a yellow oil, which was diluted with DCM (90 mL). To a solution of 4-aminobenzonitrile (2.59g, 21.92mmol) in DCM (10mL) and Et at 10 deg.C₃The resulting mixture was added dropwise to N (4.43g, 43.78 mmol). Finally the mixture was warmed to 30 ℃ and stirred for 12 h. The mixture was diluted with DCM (100mL), HCl (100mL), NaHCO₃(50mL)、H₂O (50mL) and brine (50 mL). The organic layer was washed with Na₂SO₄Dried, concentrated, and purified by silica gel column chromatography (PE: EtOAc ═ 5:1)To give 2-bromo-N- (4-cyanophenyl) benzamide (4.7g, yield: 72%) as a white solid.

Synthesis scheme J: general procedure for Compound J

To a solution of 2- (morpholinosulfonyl) aniline (1.0eq) in DMF was added carboxylic acid sm-13(1.0eq), DIPEA (1.5eq) and HATU (1.3 eq). The resulting mixture was stirred at 10-15 ℃ for 16-24 h. TLC indicated that the starting material had disappeared. The reaction mixture was concentrated, and the solid was purified by silica gel column chromatography to give compound J.

Selected compounds in table 1 were obtained using synthesis scheme J.

Synthesis scheme K-1: experimental samples for Compound 193

Methyl 2-isocyanatobenzoate (200mg, 1.13mmol) and 1- (3-aminophenyl) ethanone (167mg, 1.24mmol) were dissolved in THF (2.5ml) and heated at 100 ℃ for 15min using microwaves. The reaction mixture was saturated NaHCO₃Washed with aqueous solution and purified by column chromatography (EtOAc: hexane) to give the final product 193(262mg, yield: 75%) by¹H NMR and LCMS.

The selected compounds in table 1 were obtained using similar conditions to the reaction scheme given above. In these reactions DIPEA was used instead of TEA and the temperature was raised to 120 ℃. The reaction yield based on isolated product ranged from 47-90%.

Synthesis scheme K-2: experimental sample for Compound 196

Compound 195 was dissolved in THF (2mL) and 1N NaOH (1 mL). The reaction mixture was stirred at room temperature for 15 h. The mixture was diluted with EtOAc (20mL) and then 1N HCl (3mL) was added dropwise with constant stirring. The organic layer was extracted, dried, and concentrated. The crude product was recrystallized from EtOAc to afford compound 196(25 mg).

The selected compounds in table 1 were obtained using similar conditions to the reaction scheme given above. The reaction yield based on the isolated product ranged from 80 to 90%.

Synthesis scheme L: general procedure for Compound L

To a solution of 2- (methylsulfonamido) benzoic acid (1.0eq) in DMF were added sm-14(1.0eq), DIPEA (1.5eq) and HATU (1.3 eq). The mixture is stirred at 10-15 ℃ for 16-24 h. After completion of the reaction was indicated by TLC, the reaction mixture was concentrated and the solid was purified by silica gel chromatography to give compound L.

Selected compounds in table 1 were obtained using synthesis scheme L.

Synthetic scheme for compound 28:

a solution of compound I (320g, 1.1mmol) in EtOH (10mL) was added NaOEt (571.6mg, 8.4mmol) dropwise over 3h at 0 ℃. The reaction was acidified with 1N HCl and the solvent was removed to give the crude product. The residue was purified by preparative HPLC (0.1% TFA as additive) and most of the CH was removed by evaporation under reduced pressure₃CN and removing the remaining solvent by lyophilization to obtainCompound 28(17mg, 6% yield) as a white solid. LCMS: found 255.0[ M + H ]]。¹H NMR(400MHz,CDCl₃):7.50-7.72(m,4H),4.85(br s,1H),3.90(t,2H,J＝7.2Hz),2.53-2.72(m,5H),2.14-2.28(m,2H)。

Intermediate sm-15 was prepared by the following procedure.

4-amino-N-methylbenzenesulfonamide (200mg, 1.1mmol) and Na were added at 0 deg.C₂HPO₄(300mg, 2.2mmol) in CHCl₃To the solution in (10mL) was added 4-chlorobutyryl chloride (151mg, 1.1mol) dropwise. After the addition of the reagents, the mixture was stirred at room temperature. The mixture was concentrated to give crude sm-15, which was used in the next step without further purification.

Synthetic scheme for compound 47:

2- (4-aminophenyl) ethanol (300mg, 2.2mmol), isochroman-1, 3-dione (355mg, 2.2mmol) and KAl (SO)₄)₂-12H₂A solution of O (522mg, 11mmol) in MeCN (10mL) was stirred at room temperature for 1-1.5 h. The solvent was removed to obtain the crude product. The residue was purified by preparative HPLC (0.1% TFA as additive). The solvent was removed by evaporation under reduced pressure and lyophilized to give compound 47 as a white solid (25mg, 5.5% yield). LCMS: found 300.1[ M + H]。

Biological examples

Example 1:human chondrocyte differentiation assay

Human MSCs (50,000) were seeded into each well of a 96-well plate and cultured overnight. The compounds (in DMSO solution) were added to the cells at a final concentration of 1. mu.M, and at 5% CO₂The cells were cultured at 37 ℃ for 7 days. Cells were fixed with 10% formalin solution for 10min at room temperature and immunostained using specific antibodies against collagen type II (Abcam), Sox9(Santa Cruz) and cartilage oligomeric matrix protein (COMP, Santa Cruz) and a fluorescently labeled secondary antibody (Li-Cor). The total intensity of staining was measured using the Oddyssey CLx imaging system (Li-Cor). Vehicle (DMSO) was used as a control to determine the basal level of chondrocyte differentiation. Compounds showing an increase in staining intensity of 30% or more compared to the vehicle control were selected as active hits (active hits). Representative data are shown in table 1[ a: increased staining intensity compared to vehicle control>50 percent; b: compared with the carrier control, the staining intensity is increased by 30-50 percent]。

TABLE 1

Example 2:cell viability assay

Human MSCs, chondrocytes, osteoblasts and synoviocytes were seeded into 384-well plates at a density of 10,000 cells per well. The compound was added at a final concentration of 100. mu.M. The cells were cultured for 48 h. Cell viability was analyzed by the Cell Titer-glo (promega) assay using an EnVision plate reader (PerkinElmer). Apoptotic activity was analyzed by caspase 3/7-glo (promega) assay using EnVision plate reader (PerkinElmer).

Example 3:PK Studies in rats via intra-articular injection

Mu.l of compound solution (100. mu.M in PBS containing 0.1% DMSO) was injected into the joint space of the right knee of each rat. Animals were bled at 1,3, 4,6, 7, 8, 9 and 10 hours post injection. Animals were sacrificed 2 or 12 hours after dosing. Injected plasma and joint lavage fluid of the knee were collected. The amount of injected compound was analyzed using LCMS.

Example 4:rat Medial Meniscal Tear (MMT) Osteoarthritis (OA) model

The medial meniscus of the right knee of each animal was surgically torn to induce OA. Administration of compound solution (30 μ l, 100 μ M in PBS containing 0.1% DMSO) was initiated 7 days post-surgery, once weekly for three weeks. Body weight and gait deficits were monitored weekly just prior to dosing. Animals were sacrificed on day 28 post-surgery. The knee joint was treated by surgery, and the cartilage was histochemically stained and evaluated.

After 4-6 days in 5% formic acid decalcifying agent, the operated joint was frontal cut into two approximately equal halves and embedded in paraffin. Three sections were cut from each of the right operated knees (g1-8) in steps of approximately 200 μm and stained with toluidine blue. Single sections were prepared and stained with toluidine blue for the left knee of group 1 and the right knee of group 9.

All three sections of each operated knee were analyzed by microscopy. The worst case for both halves on each slide was determined for total cartilage degradation, proteoglycan loss, collagen damage and osteophyte formation. The values for each parameter are then averaged across the three slices to determine the overall subjective score.

In addition, for some parameters (noted below), regional differences across the tibial plateau were considered by dividing each slice into three zones (1-outer, 2-middle, 3-inner). In the surgical OA model, the outer (z1) and middle (z2) thirds were most severely affected, while there was a minor change in the inner third (z 3). When scoring a band alone, a score is assigned based on the percentage of area of the affected band. Zone areas were delineated using an ocular micrometer.

The following parameters were measured and/or scored:

total cartilage degradation includes important parameters of chondrocyte death/loss, proteoglycan loss, and collagen loss or fibrillation. Cartilage degradation in the tibia was scored from none to severe (values 0-5) for each zone using the following criteria:

0-no degradation

1-minimal degeneration, within this zone 5-10% of the matrix appears non-viable due to significant chondrocyte loss (greater than 50% of normal cell density). PG loss is usually present in these areas of cell loss, and there may be collagen matrix loss.

2-mild degeneration, within this zone, 11-25% of the matrix appeared non-viable due to significant chondrocyte loss (greater than 50% of normal cell density). PG loss is usually present in these areas of cell loss, and there may be collagen matrix loss.

3-moderate degeneration, within this zone 26-50% of the matrix appeared non-viable due to significant chondrocyte loss (greater than 50% of normal cell density). PG loss is usually present in these areas of cell loss, and there may be collagen matrix loss.

4-clear degeneration, within this zone 51-75% of the matrix appeared non-viable due to significant chondrocyte loss (greater than 50% of normal cell density). PG loss is usually present in these areas of cell loss, and there may be collagen matrix loss.

5-severe degeneration, within this zone 76-100% of the matrix appeared non-viable due to significant chondrocyte loss (greater than 50% of normal cell density). PG loss is usually present in these areas of cell loss, and there may be collagen matrix loss.

In some cases, image analysis may be used to determine the exact matrix viability and/or loss% in each zone or selected zones, such that absolute% may be compared rather than scores (0-5). In addition to the data representing each zone, the 3-zone sum of cartilage degradation was calculated.

The same procedure applies to the evaluation of femoral cartilage, except that the lesions are not analyzed on a zonal basis, as the lesions are typically not distributed over the entire surface in a zonal pattern. The total width of the load bearing surface (about 2000 μm for femurs) was determined and the above criteria were applied to 1/3, 2/3 or 3/3, which were most severely affected. For example, if 1/3 (lesions may be in the center of a plateau covering about 667 μm) of the total area had minimal degeneration (5-10% of the total area had loss of chondrocytes and/or stroma), a score of 1 was assigned. If the minimum degradation extends over the entire surface (3/3), a score of 3 is assigned. If the entire femoral cartilage is not present due to severe diffuse degradation, a score of 15 is obtained.

In addition to this overall cartilage degradation score, collagen matrix damage alone was scored to determine more specific effects of the agent. Collagen damage across the medial tibial plateau (the most severely affected sections of the two halves) was quantified by measuring the total width of:

random injury (fibrillation of superficial to full thickness loss)

Severe damage (total or near total loss of collagen to the tidal mark (> 90% thickness))

Evident lesions (extending through 61-90% of the thickness of the cartilage)

Moderate lesions (extending through 31-60% of the cartilage thickness)

Mild lesions (extending through 11-30% of the thickness of the cartilage)

Minimal damage (extremely superficial, affecting only the upper 10%)

In addition to the subjective overall cartilage score above, two cartilage degradation width measurements were also performed:

·total tibial cartilage degradation Width (μm)Is a micrometer measurement of the overall degree to which the tibial plateau is affected by any type of degeneration (cell loss, proteoglycan loss, or collagen damage). This measurement extends across the surface from the start of an osteophyte with adjacent cartilage degradation (exterior 1/3) to the point where the tangential layer and underlying cartilage appear histologically normal.

·Width of parenchymal cartilage degradation (μm)Reflecting chondrocytes and proteoglycansThe loss extends through more than 50% of the cartilage thickness in the area of tibial cartilage degradation. Typically, for this parameter, collagen damage is mild (25% depth) or greater, but chondrocyte and proteoglycan loss extends to at least 50% or greater of the cartilage depth.

The micrometer depth of any type of lesion (loss of both chondrocytes and proteoglycans, but may have good retention of collagen matrix and no fibrillation), expressed as the ratio of varying region depth to the tidemark, was obtained in the region of greatest lesion severity in each of the three zones across the tibial surface at the mid-point of the zone. This measurement is the most critical analysis for any type of microscopic change that is present. The denominator can serve as an average measure of cartilage thickness for each of the three zones when measured at the midpoint of the zones for comparison of anabolism.

Scoring and classification of osteophytes (classification as small, medium and large) was performed using an ocular micrometer. In order to be able to measure and specify osteophytes, the marginal zone proliferation change must be > 200 μm. The largest osteophytes in each slice (typically found in the tibia) were assigned a score according to the following criteria:

1 is small, up to 299 μm

2 medium, 300-

Large 3 ═ 400-

Maximum 4, 500-

5 is extremely large, not less than 600

Actual osteophyte measurements (tidal mark to the furthest point extending towards the synovium) were also recorded.

The sum of the three zone bands (average of three levels) of the femoral cartilage degradation score and the tibial cartilage degradation score were added to formTotal cartilage degradation score. Add the average osteophyte score per joint to this value to generateTotal joint score.

Image analysis

To quantify and compare cartilage matrix retention, cartilage area measurements were made from the most severely affected sections of each animal. Micrographs were taken with a CoolSNAP-Pro microscopy camera and loaded into ImagePro Plus software. From the tracings of these micrographs, the following measurements were made, four per page, which were included in the report:

total area from tidal mark to surface (or surface convex in degenerated area) on 9cm (photomicrograph) tibial plateau, measured from the inner edge of the osteophyte

Non-viable matrix (cartilage with less than 50% chondrocytes, proteoglycans and intact collagen) and matrix-free area within the total area

Area of total area without matrix

The area of non-viable matrix was subtracted from the total area to obtain the area of viable matrix, and the area without matrix was subtracted from the total area to obtain the area of any matrix (collagen matrix with or without chondrocytes and proteoglycans). These two values are then compared back to the total area to yieldPercentage of active matrix areaAndany percentage of substrate areaThey were compared between groups. Five left knees from the vector group were included as normal controls in this procedure. This process can be used to analyze the entire surface or selected zones based on the severity of the lesion and the apparent therapeutic effect.

Synovial responses, if abnormal, are described (should be primarily fibrotic) and characterized in terms of type and extent of inflammation, but are not included in the OA score.

Lesions of calcified cartilage layers and subchondral bone (worst case for all sections) were scored using the following criteria:

0 is unchanged

1-basophilia increased at the tidal scale, with no disruption of the tidal scale, no bone marrow alteration or minimal and focal if present

Increased basophilia at the tidal scale, minimal to mild focal fragmentation of calcified cartilage at the tidal scale, interstitial changes in bone marrow involving 1/4 in total area, but usually limited to the subchondral area under the lesion

3-basophilia increased at the tidal scale, mild to marked focal or multifocal fragmentation of calcified cartilage (multifocal), interstitial changes in bone marrow involving up to 3/4 of total area, areas of bone marrow chondrogenesis that may be significant, but articular cartilage not largely collapsed into epiphyseal bone (well-defined depression in the surface)

4-basophilia increased at the tidal scale, pronounced to severe fragmentation of calcified cartilage, marrow stromal changes involving up to 3/4 in area, and the articular cartilage having collapsed into epiphyses at a depth of 250 μm or less from the tidal scale (clear pits visible in the superficial cartilage)

Increased basophilia at the tidal scale, marked to severe fragmentation of calcified cartilage, bone marrow stromal changes involving up to 3/4 areas, and the articular cartilage having collapsed into epiphyses, greater than 250 μm deep from the tidal scale

In addition, the thickness of medial synovial fluid/collateral ligament repair in the non-tangential region of the slice was measured.

All knees were measured on the medial and lateral sides (2 measurements/joint), and growth plane thickness was measured at the approximate midpoint of the medial and lateral epiphyses (assuming a non-tangential region of the slice).

Example 5:extraction and quantification of chondrogenic compounds in articular and plasma rat samples

LC-MS/MS analysis for chondrogenic compounds was performed using an API 3000 equipped with an Agilent 1100HPLC and a leap technologies autosampler. An HPLC Phenomenex 5 micron, 100A LunaC18(2) analytical column was used, with dimensions of 2.0X 50mm (cat # 00B-4252-B0), a temperature of 30 ℃, a flow rate of 0.6mL/min, an injection volume of 10uL, and a run time of 6.0 min. Mobile phase a1 was 0.1% aqueous formic acid and mobile phase B1 was 0.1% formic acid in acetonitrile. Gradient 90% A1/10% B1 at time 0; 90% A1/10% B1 at 1.0 min; at time 2.0min, 10% A1/90% B1; at time 4.0min, 10% A1/90% B1; at time 4.10min, 90% A1/10% B1; the time at 6.0min was 90% A1/10% B1. Quantification of analytes and internal standards was performed using Multiple Reaction Monitoring (MRM) quantification. Specific methods used to administer and measure exposure in plasma and the concentrations observed in joint extracts are listed below.

Rat plasma samples: a calibration standard curve was prepared by serial dilution of concentrated, spiked compound solutions in control rat plasma. Calibration standards and rat plasma samples were prepared by protein precipitation by adding acetonitrile and an aliquot of the internal standard to each aliquot of standard and sample. After vortex mixing and centrifugation, aliquots of the supernatant from each standard and sample were diluted with aqueous formic acid, mixed and injected. All plasma samples collected after IA administration (starting at t ═ 0, 0.5, 1,2, 4, and 6 h) indicated no systemic exposure to any of the compounds listed in table 2.

Rat knee joint samples: a calibration standard curve was prepared by serial dilution of concentrated, spiked compound solutions in internal standard diluent. The internal standard diluent is prepared by dissolving an internal standard compound in acetonitrile at a specific concentration. Rat knee samples at each time point were crushed individually and transferred to each centrifuge tube and 1.0-mL of internal standard diluent was added. Each tube was vortexed and centrifuged for 30 minutes. The supernatant was removed from each tube and injected onto the column for analysis. In addition, plasma samples were obtained by retroorbital bleeding into heparin-coated tubes and stored at-80 ℃ and later processed similarly to the protocol described above for rat plasma samples.

Compound administration and tissue treatment: mu.L of a 100. mu.M solution of the compound (PBS containing 0.1% DMSO) was injected into the intra-articular space of the right posterior knee of each animal. Animals were euthanized at the indicated time points (0hr, 0.5hr, 1hr, 2hr, 4hr, and 6 hr). Four animals were used at each time point. The injected knee joints were harvested and snap frozen in liquid nitrogen. The whole joint was ground to a powder under refrigeration, mixed with 1mL acetonitrile containing an internal standard and incubated overnight at 4 ℃, vortexed, and centrifuged for 30 min. The supernatant of each sample was analyzed using LC-MS/MS. The data shown in table 2 represent the concentrations observed in the knee extract. ND was not determined.

TABLE 2

Example 6: parenteral compositions of the compounds presented herein

To prepare a parenteral pharmaceutical composition suitable for administration by injection, 100mg of a compound set forth herein or a water-soluble pharmaceutically acceptable salt thereof is dissolved in DMSO and then mixed with 10ml of a 0.9% sterile saline solution. The mixture is combined into dosage units suitable for administration by injection.

Example 7: oral compositions of the compounds presented herein

To prepare a pharmaceutical composition for oral delivery, 400mg of a compound presented herein and the following ingredients are intimately mixed and compressed into a single scored tablet.

Tablet formulation

The following ingredients were intimately mixed and loaded into hard shell gelatin capsules.

Capsule formulation

Claims

1. Use of a compound of formula IIc, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, in the manufacture of a medicament for ameliorating arthritis or joint damage or inducing differentiation of mesenchymal stem cells into chondrocytes in a mammal:

wherein

n is 0, 1,2, 3 or 4;

b is NHSO₂R³、NR³SO₂R³、NHSO₂R⁴Or NR³SO₂R⁴；

R²Is mono-or di-substituted phenyl, said substituents being independently selected from optionally substituted alkyl, halo and C (O) alkyl;

R⁴is optionally substituted phenyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, optionally substituted aralkyl or optionally substituted alkyl;

A is C (O) NHR²Or C (O) NR²R⁴。

2. The use of claim 1, wherein B is NHSO₂R³Or NR³SO₂R³。

3. The use of claim 2, wherein R³Is an optionally substituted alkyl group.

4. The use of claim 3, wherein R³Is CH₃。

5. The use of claim 1, wherein B is NHSO₂R⁴Or NR³SO₂R⁴。

6. The use of claim 5, wherein R⁴Is optionally substituted phenyl.

7. The method of claim 1, wherein A is C (O) NHR²。

8. The use of claim 1, wherein n is 0.

9. The use of claim 1, wherein n is 1.

10. The use of claim 1, wherein R¹Independently selected from Cl, F, CH₂OH、CH₂NH₂、OCH₃、OCF₃、OCHF₂、CN、NO₂、CO₂H and CO₂CH₃。

11. The use of claim 1, wherein the compound is selected from

12. A compound selected from

Or pharmaceutically acceptable thereofA salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer.

13. A pharmaceutical composition comprising the compound of claim 112, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer thereof, and a pharmaceutically acceptable excipient.

14. The pharmaceutical composition of claim 13, further comprising an additional compound that is therapeutically effective for treating arthritis or joint damage and/or a symptom associated with arthritis or joint damage in a mammal.

15. The pharmaceutical composition of claim 14, wherein the additional compound is selected from NSAIDS, analgesics, angiopoietin-like 3 protein (ANGPTL3) or chondrogenic variants thereof, oral salmon calcitonin, iNOS inhibitors, vitamin D3, caspase inhibitors, collagen hydrolysates, FGF18, BMP7, Avocado Soybean Unsaponifiables (ASU), and hyaluronic acid.

16. Use of a compound of formula I, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer composition thereof, in the manufacture of a medicament for ameliorating arthritis or joint damage in a mammal:

wherein

n is 0, 1,2, 3 or 4;

m is 1,2, 3 or 4;

b is CO₂R⁴、CH₂CO₂H、CH₂CO₂R⁴Or optionally substituted phenyl;

X is O or CR⁵R⁶；

Each R³Independently selected from H, CN, halo, C (O) R⁴、CO₂H、CO₂R⁴、C(O)NR⁴R¹¹Alkyl, optionally substituted alkoxy、SO₂R⁴、(CR⁷R⁸)OR⁴、(CR⁷R⁸)NR⁴R¹¹、(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)NR⁴R¹¹、(CR⁷R⁸)C(O)R⁴、(CR⁷R⁸)C(O)OR⁴、(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)C(O)R⁴、X(CR⁷R⁸)C(O)OR⁴、X(CR⁷R⁸)C(O)NR⁴R¹¹、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)R⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)OR⁴、X(CR⁷R⁸)(CR⁹R¹⁰)C(O)NR⁴R¹¹、(CR⁷R⁸)NR⁴SO₂R⁴And C (═ NOR)⁴)R⁴；

Each R⁴Independently selected from H and optionally substituted alkyl;

Provided that

b) The compound is not selected from

17. Use of a compound of formula III, or a pharmaceutically acceptable salt, solvate, polymorph, prodrug, ester, metabolite, N-oxide, stereoisomer, or isomer composition thereof, in the manufacture of a medicament for ameliorating arthritis or joint damage in a mammal:

wherein

n is 0, 1,2, 3 or 4;

x is O, NH or NR⁶；

A is C (O), CH₂Or CH-CR³R⁴-C(O)R²；

R²Is optionally substituted aryl or optionally substituted heteroaryl;

each R³And R⁴Independently is H or optionally substituted alkyl;

R⁶Is optionally substituted phenyl;

provided that

a) If A is CH-CR³R⁴-C(O)R²X is O or NH;

b) if n is 0, A is CHCH₂C(O)R²And X is O, then R²Is not provided withOrAnd is

c) If A is C (O) or CH₂X is NR⁶And R is⁶Is not provided with