CN102485720A - heat shock protein 90 inhibitor - Google Patents

Abstract

Provided herein are heat shock protein 90 inhibitor compounds, e.g., compounds of formula I, pharmaceutical compositions comprising the compounds, and methods of making the same. Methods of their use for treating proliferative diseases are also provided.

Description

heat shock protein 90 inhibitor

technical field

Provided herein are heat shock protein 90 inhibitor compounds, pharmaceutical compositions comprising such compounds, and methods of making them. Also provided are methods of their use in the treatment of proliferative diseases.

Background technique

Heat shock protein 90 (Heat shock protein 90, HSP90) is a highly conserved intracellular molecular chaperone, which regulates the post-translational folding of client proteins (Welch and Feramiscom J.Biol.Chem.1982, 257, 14949-14959 ). HSP90 exerts its chaperone function to ensure the correct conformation, activation, intracellular localization and proteolytic turnover of a series of proteins involved in cell growth, differentiation and survival (Maloney and Workman, Expert Opin. Biol. Ther. 2002, 2, 3 -24; and Whitesell and Lindquist, Nature Reviews Cancer 2005, 5, 761-772). HSP90 is essential for the stability and function of many oncogenic client proteins including ERBB2, BCR-ABL, AKT/PKB, C-RAF, CDK4, PLK-1, MET, mutant p53, HIF-1α , steroid hormone receptors (estrogen and androgen) and telomerase hTERT (Maloney and Workman, Expert Opin. Biol. Ther. 2002, 2, 3-24).

HSP90 expression and activation are often upregulated in tumor cells. HSP90 in tumor cells has been shown to exist in a hyperactive state with elevated ATPase activity, which is highly sensitive to HSP90 inhibition compared to the abundant latent form found in normal cells (Richter et al., Nat. Struct. Mol. Biol. 2007, 14, 90-94). This suggests that HSP90 inhibitors can be used to selectively target tumor cells. Furthermore, since some oncogenic client proteins are associated with multiple hallmark features of malignancy such as deregulated signal transduction, cell cycle progression, apoptosis, immortalization, angiogenesis, invasion, and metastasis, inhibition of HSP90 has important It would be desirable to provide effective combined blockade of tumor cells (Hanahan and Weinberg, Cell 2000, 100, 57-70; and Workman, Cancer Lett. 2004, 206, 149-157). Inhibition of HSP90 has been shown to cause degradation of client proteins through the ubiquitin-proteasome pathway, which leads to simultaneous depletion of multiple oncoproteins and combinatorial downregulation of signals propagated through many oncogenic signaling pathways (Connell et al., Nature Cell Biology 2001, 3, 93-96; Demand et al., Current Biology 2001, 11, 1569-1577; Maloney and Workman, Expert Opin. Biol. Ther. 2002, 2, 3-24; and Workman, Cancer Lett. 2004 , 206, 149-157).

For example, the small molecule HSP90 inhibitor 17-allyamino-17-demethoxygeldanamycin (17-AAG), which binds ATP at the _NH2 -terminal region of HSP90 binding site, and inhibit its ATPase activity, the small molecule HSP90 inhibitor has been shown to effectively inhibit a variety of cancers in cancer cell lines and human tumor xenograft models (Kelland et al., J.Natl.Cancer Inst.1999 , 91, 1940-1949; Solit et al., Clin. Cancer Res.2002, 8, 986-993; Jones et al., Blood, 2003, 103, 1855-1861; Price et al., Cancer Res.2005, 65, 4929-4938; and Banerji et al., Clin. Cancer Res. 2005, 11, 7023-7032). Inhibition of the basic ATPase activity of HSP90 leads to downregulation of multiple client proteins through the ubiquitin-proteasome pathway, resulting in cell cycle arrest and apoptosis (Schulte et al., Biochem.Biophys.Res.Commun.1997, 239, 655-659; and Hostein et al., Cancer Res. 2001, 61, 4003-4009). Therefore, there is a need for HSP90 inhibitors that are useful in the effective treatment of proliferative diseases such as cancer.

Contents of the invention

Summary of the invention

或单一对映异构体、对映异构体的混合物或非对映异构体的混合物；或其药学可接受的盐、溶剂合物或水合物；其中：R¹是氢、C_1-6烷基、C_2-6烯基、C_2-6炔基、C_3-7环烷基、C_6-14芳基、C_7-15芳烷基、杂芳基或杂环基；R²是杂芳基或杂芳基-C_1-6烷基；R⁵是(i)氢、卤素、氰基或硝基；(ii)C_1-6烷基、C_2-6烯基、C_2-6炔基、C_3-7环烷基、C_6-14芳基、C_7-15芳烷基、杂芳基或杂环基；或(iii)-C(O)R^1a、-C(O)OR^1a、-C(O)NR^1bR^1c、-C(NR^1a)NR^1bR^1c、-OR^1a、-OC(O)R^1a、-OC(O)OR^1a、-OC(O)NR^1bR^1c、-OC(＝NR^1a)NR^1bR^1c、-OS(O)R^1a、-OS(O)₂R^1a、-OS(O)NR^1bR^1c、-OS(O)₂NR^1bR^1c、-NR^1bR^1c、-NR^1aC(O)R^1d、-NR^1aC(O)OR^1d、-NR^1aC(O)NR^1bR^1c、-NR^1aC(＝NR^1d)NR^1bR^1c、-NR^1aS(O)R^1d、-NR^1aS(O)₂R^1d、-NR^1aS(O)NR^1bR^1c、-NR^1aS(O)₂NR^1bR^1c、-SR^1a、-S(O)R^1a、-S(O)₂R^1a、-S(O)NR^1bR^1c或-S(O)₂NR^1bR^1c；和各个R^1a、R^1b、R^1c和R^1d独立地是氢、C_1-6烷基、C_2-6烯基、C_2-6炔基、C_3-7环烷基、C_6-14芳基、C_7-15芳烷基、杂芳基或杂环基；或R^1b和R^1c与它们连接的N原子一起形成杂芳基或杂环基；其中各个烷基、烯基、炔基、环烷基、芳基、芳烷基、杂环基、杂芳基和杂芳基-烷基任选被一个或多个基团Q取代，各个Q独立地选自(a)氰基、卤素和硝基；(b)C_1-6烷基、C_2-6烯基、C_2-6炔基、C_3-7环烷基、C_6-14芳基、C_7-15芳烷基、杂芳基和杂环基，各自任选被一个或多个(在一个实施方案中，为1、2、3或4个)取代基Q^a取代；和(c)-C(O)R^a、-C(O)OR^a、-C(O)NR^bR^c、-C(NR^a)NR^bR^c、-OR^a、-OC(O)R^a、-OC(O)OR^a、-OC(O)NR^bR^c、-OC(＝NR^a)NR^bR^c、-OS(O)R^a、-OS(O)₂R^a、-OS(O)NR^bR^c、-OS(O)₂NR^bR^c、-NR^bR^c、-NR^aC(O)R^d、-NR^aC(O)OR^d、-NR^aC(O)NR^bR^c、-NR^aC(＝NR^d)NR^bR^c、-NR^aS(O)R^d、-NR^aS(O)₂R^d、-NR^aS(O)NR^bR^c、-NR^aS(O)₂NR^bR^c、-SR^a、-S(O)R^a、-S(O)₂R^a、-S(O)NR^bR^c和-S(O)₂NR^bR^c，其中各个R^a、R^b、R^c和R^d独立地是(i)氢；(ii)C_1-6烷基、C_2-6烯基、C_2-6炔基、C_3-7环烷基、C_6-14芳基、C_7-15芳烷基、杂芳基或杂环基，各自任选被一个或多个(在一个实施方案中，为1、2、3或4个)取代基Q^a取代；或(iii)R^b和R^c与它们连接的N原子一起形成杂环基，任选被一个或多个(在一个实施方案中，为1、2、3或4个)取代基Q^a取代；其中各个Q^a独立地选自(a)氰基、卤素和硝基；(b)C_1-6烷基、C_2-6烯基、C_2-6炔基、C_3-7环烷基、C_6-14芳基、C_7-15芳烷基、杂芳基和杂环基；和(c)-C(O)R^e、-C(O)OR^e、-C(O)NR^fR^g、-C(NR^e)NR^fR^g、-OR^e、-OC(O)R^e、-OC(O)OR^e、-OC(O)NR^fR^g、-OC(＝NR^e)NR^fR^g、-OS(O)R^e、-OS(O)₂R^e、-OS(O)NR^fR^g、-OS(O)₂NR^fR^g、-NR^fR^g、-NR^eC(O)R^h、-NR^eC(O)OR^f、-NR^eC(O)NR^fR^g、-NR^eC(＝NR^h)NR^fR^g、-NR^eS(O)R^h、-NR^eS(O)₂R^h、-NR^eS(O)NR^fR^g、-NR^eS(O)₂NR^fR^g、-SR^e、-S(O)R^e、-S(O)₂R^e、-S(O)NR^fR^g和-S(O)₂NR^fR^g；其中各个R^e、R^f、R^g和R^h独立地是(i)氢；(ii)C_1-6烷基、C_2-6烯基、C_2-6炔基、C_3-7环烷基、C_6-14芳基、C_7-15芳烷基、杂芳基或杂环基；或(iii)R^f和R^g与它们连接的N原子一起形成杂环基。Provided herein are compounds of Formula I:

Or a single enantiomer, a mixture of enantiomers or a mixture of diastereomers; or a pharmaceutically acceptable salt, solvate or hydrate thereof; wherein: R ¹ is hydrogen, C _{1- 6} alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 aryl, C _7-15 aralkyl, heteroaryl or heterocyclic; R ² is heteroaryl or heteroaryl-C _1-6 alkyl; R ⁵ is (i) hydrogen, halogen, cyano or nitro; (ii) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 aryl, C _7-15 aralkyl, heteroaryl or heterocyclyl; or (iii)-C(O)R ^1a , -C(O)OR ^1a , -C(O)NR ^1b R ^1c , -C(NR ^1a )NR ^1b R ^1c , -OR ^1a , -OC(O)R ^1a , -OC(O)OR ^1a , - OC(O)NR ^1b R ^1c , -OC(=NR ^1a )NR ^1b R ^1c , -OS(O)R ^1a , -OS(O) ₂ R ^1a , -OS(O)NR ^1b R ^1c , -OS (O) ₂ NR ^1b R ^1c , -NR ^1b R ^1c , -NR ^1a C(O)R ^1d , -NR ^1a C(O)OR ^1d , -NR ^1a C(O)NR ^1b R ^1c , -NR ^1a C(=NR ^1d )NR ^1b R ^1c , -NR ^1a S(O)R ^1d , -NR ^1a S(O) ₂ R ^1d , -NR ^1a S(O)NR ^1b R ^1c , -NR ^1a S(O ) ₂ NR ^1b R ^1c , —SR ^1a , —S(O)R ^1a , —S(O) ₂ R ^1a , —S(O)NR ^1b R ^1c , or —S(O) ₂ NR ^1b R ^1c ; and Each of R ^1a , R ^1b , R ^1c and R ^1d is independently hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 Aryl, C _7-15 aralkyl, heteroaryl or heterocyclic group; or R ^1b and R ^1c form heteroaryl or heterocyclic group together with their connected N atoms; wherein each alkyl, alkenyl, alkyne radical, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, and heteroaryl-alkyl are optionally substituted by one or more groups Q, each Q independently selected from (a) cyano , halogen and nitro; (b) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 aryl, C _7-15 aromatic Alkyl, heteroaryl and heterocyclyl, each optionally substituted by one or more (in one embodiment, 1, 2, 3 or 4) substituent Q ^a ; and (c)-C(O )R ^a , -C(O)OR ^a , -C(O)NR ^b R ^c , -C(NR ^a )NR ^b R ^c , -OR ^a , -OC(O)R ^a , -OC(O)OR ^a , -OC(O)NR ^b R ^c , -OC(=NR ^a )NR ^b R ^c , -OS(O)R ^a , -OS(O) ₂ R ^a , -OS(O)NR ^b R ^c , -OS(O) ₂ NR ^b R ^c , -NR ^b R ^c , -NR ^a C(O)R ^d , -NR ^a C(O)OR ^d , -NR ^a C(O)NR ^b R ^c , -NR ^a C(=NR ^d ) NR ^b R ^c , -NR ^a S(O)R ^d , -NR ^a S(O) ₂ R ^d , -NR ^a S(O)NR ^b R ^c , -NR ^a S(O) ₂ NR ^b R ^c , -SR ^a , -S(O)R ^a , -S(O) ₂ R ^a , -S(O)NR ^b R ^c and -S(O) ₂ NR ^b R ^c , wherein each of R ^a , R ^b , R ^c and R ^d are independently (i) hydrogen; (ii) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 Aryl, C _aralkyl , heteroaryl or heterocyclyl, each optionally substituted by one or more (in one embodiment, 1, 2, 3 or 4) substituents ^Qa ; or (iii) R ^b and R ^c together with the N atom to which they are attached form a heterocyclic group, optionally substituted by one or more (in one embodiment, 1, 2, 3 or 4) substituent Q ^a ; wherein each Q ^a is independently selected from (a) cyano, halogen and nitro; (b) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkane base, C _6-14 aryl, C _7-15 aralkyl, heteroaryl and heterocyclyl; and (c)-C(O)R ^e , -C(O)OR ^e , -C(O) NR ^f R ^g , -C(NR ^e )NR ^f R ^g , -OR ^e , -OC(O)R ^e , -OC(O)OR ^e , -OC(O)NR ^f R ^g , -OC(= NR ^e ) NR ^f R ^g , -OS(O)R ^e , -OS(O) ₂ R ^e , -OS(O)NR ^f R ^g , -OS(O) ₂ NR ^f R ^g , -NR ^f R ^g , -NR ^e C(O)R ^h , -NR ^e C(O)OR ^f , -NR ^e C(O)NR ^f R ^g , -NR ^e C(=NR ^h )NR ^f R ^g , -NR ^e S(O)R ^h , -NR ^e S(O) ₂ R ^h , -NR ^e S(O)NR ^f R ^g , -NR ^e S(O) ₂ NR ^f R ^g , -SR ^e , -S(O)Re ^e , -S(O) ₂ R ^e , -S(O)NR ^f R ^g and -S(O ) ₂ NR ^f R ^g ; wherein each R ^e , R ^f , R ^g and ^Rh are independently (i) hydrogen; (ii) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkyne group, C _3-7 cycloalkyl group, C _6-14 aryl group, C _7-15 aralkyl group, heteroaryl group or heterocyclic group; or (iii) R ^f and R ^g form together with the N atom to which they are attached heterocyclyl.

或单一对映异构体、对映异构体的混合物或非对映异构体的混合物；或其药学可接受的盐、溶剂合物或水合物；其中R¹、R²、R⁵的定义如上文式I所述。In another embodiment, the compound of formula I has the structure of formula II:

Or a single enantiomer, a mixture of enantiomers or a mixture of diastereomers; or a pharmaceutically acceptable salt, solvate or hydrate thereof; wherein R ¹ , R ² , R ⁵ Definitions are as described for formula I above.

Detailed description of the invention

The general formulas provided herein include the groups R ¹ , R ² and R ⁵ in formulas I and II as further defined herein. All combinations of the embodiments provided herein for such groups are within the scope of the present disclosure.

In certain embodiments, R ¹ is hydrogen. In certain embodiments, R ¹ is C _1-6 alkyl, optionally substituted with one or more substituents Q. In certain embodiments, R ¹ is methyl, ethyl or propyl. In certain embodiments, R ¹ is methyl, ethyl or isopropyl. In certain embodiments, R ¹ is C _2-6 alkenyl, optionally substituted with one or more substituents Q. In certain embodiments, R ¹ is C _2-6 alkynyl, optionally substituted with one or more substituents Q. In certain embodiments, R ¹ is C _3-7 cycloalkyl, optionally substituted with one or more substituents Q. In certain embodiments, R ¹ is C _6-14 aryl, optionally substituted with one or more substituents Q. In certain embodiments, R ¹ is C _7-15 aralkyl, optionally substituted with one or more substituents Q. In certain embodiments, ^R1 is heteroaryl, optionally substituted with one or more substituents Q. In certain embodiments, ^R1 is heterocyclyl, optionally substituted with one or more substituents Q.

In certain embodiments, ^R2 is heteroaryl, optionally substituted with one or more substituents Q. In certain embodiments, R ² is heteroaryl-C _1-6 alkyl, wherein the alkyl and heteroaryl are each independently and optionally substituted with one or more substituents Q. In certain embodiments, ^R is 6-membered heteroaryl-C _1-6 alkyl, wherein the alkyl and heteroaryl are each independently and optionally substituted with one or more substituents Q. In certain embodiments, R ² is 6-membered heteroaryl-methyl, wherein the heteroaryl is optionally substituted with one or more substituents Q. In certain embodiments, ^R2 is pyridylmethyl, optionally substituted with one or more substituents Q. In certain embodiments, R ² is pyridin-2-yl-methyl, pyridin-3-yl-methyl or pyridin-4-yl-methyl.

In certain embodiments, ^R is 5,6-fused bicyclic heteroaryl-C _1-6 alkyl, wherein the alkyl and heteroaryl are each independently and optionally replaced by one or more substituents Q replace. In certain embodiments, ^R is 5,6-fused bicyclic heteroaryl-methyl, wherein the heteroaryl is optionally substituted with one or more substituents Q. In certain embodiments, ^R is pyrazolo[1,5-a]pyrimidin-3-yl)methyl, optionally substituted with one or more substituents Q. In certain embodiments, R ² is chloropyrazolo[1,5-a]pyrimidinyl)methyl. In certain embodiments, R ² is 6-chloropyrazolo[1,5-a]pyrimidin-3-yl)methyl.

In certain embodiments, ^R is 6,6-fused bicyclic heteroaryl-C _1-6 alkyl, wherein the alkyl and heteroaryl are each independently and optionally replaced by one or more substituents Q replace. In certain embodiments, ^R is 6,6-fused bicyclic heteroaryl-methyl, wherein the heteroaryl is optionally substituted with one or more substituents Q. In certain embodiments, R ² is quinolinyl-methyl, optionally substituted with one or more substituents Q. In certain embodiments, R ² is quinolin-6-yl-methyl, optionally substituted with one or more substituents Q. In certain embodiments, R ² is 7-fluoro-quinolin-6-yl-methyl.

In certain embodiments, R ⁵ is hydrogen. In certain embodiments, R ⁵ is halogen. In certain embodiments, R ⁵ is fluoro, chloro, bromo or iodo. In certain embodiments, R ⁵ is chloro. In certain embodiments, R ⁵ is cyano. In certain embodiments, R ⁵ is nitro. In certain embodiments, R ⁵ is C _1-6 alkyl, optionally substituted with one or more substituents Q. In certain embodiments, R ⁵ is methyl, fluoromethyl, difluoromethyl or trifluoromethyl. In certain embodiments, R ⁵ is C _2-6 alkenyl, optionally substituted with one or more substituents Q. In certain embodiments, R ⁵ is C _2-6 alkynyl, optionally substituted with one or more substituents Q. In certain embodiments, R ⁵ is C _3-7 cycloalkyl, optionally substituted with one or more substituents Q. In certain embodiments, ^R5 is cyclopropyl, optionally substituted with one or more substituents Q. In certain embodiments, R ⁵ is C _6-14 aryl, optionally substituted by one or more substituents Q. In certain embodiments, R ⁵ is C _7-15 aralkyl, optionally substituted by one or more substituents Q. In certain embodiments, ^R5 is heteroaryl, optionally substituted with one or more substituents Q. In certain embodiments, ^R5 is heterocyclyl, optionally substituted with one or more substituents Q.

In certain embodiments, R ⁵ is -C(O)R ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -C(O)OR ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -C(O)NR ^1b R ^1c , wherein R ^1b and R ^1c are each as defined herein. In certain embodiments, R ⁵ is -C(NR ^1a )NR ^1b R ^1c , wherein each of R ^1a , R ^1b and R ^1c is as defined herein. In certain embodiments, R ⁵ is -OR ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is C _1-6 alkoxy, optionally substituted with one or more substituents as defined herein. In certain embodiments, R ⁵ is methoxy, ethoxy or propoxy. In certain embodiments, R ⁵ is -OC(O)R ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -OC(O)OR ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -OC(O)NR ^1b R ^1c , wherein R ^1b and R ^1c are each as defined herein. In certain embodiments, R ⁵ is -OC(=NR ^1a )NR ^1b R ^1c , wherein each of R ^1a , R ^1b and R ^1c is as defined herein. In certain embodiments, R ⁵ -OS(O)R ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -OS(O) ₂ R ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -OS(O)NR ^1b R ^1c , wherein R ^1b and R ^1c are each as defined herein. In certain embodiments, R ⁵ is -OS(O) ₂ NR ^1b R ^1c , wherein R ^1b and R ^1c are each as defined herein. In certain embodiments, R ⁵ is -NR ^1b R ^1c , wherein R ^1b and R ^1c are each as defined herein. In certain embodiments, R ⁵ is -NR ^1a C(O)R ^1d , wherein R ^1a and R ^1d are each as defined herein. In certain embodiments, R ⁵ is -NR ^1a C(O)OR ^1d , wherein R ^1a and R ^1d are each as defined herein. In certain embodiments, R ⁵ is -NR ^1a C(O)NR ^1b R ^1c , wherein each of R ^1a , R ^1b and R ^1c is as defined herein. In certain embodiments, R ⁵ is -NR ^1a C(=NR ^1d )NR ^1b R ^1c , wherein each of R ^1a , R ^1b , R ^1c and R ^1d is as defined herein. In certain embodiments, R ⁵ is -NR ^1a S(O)R ^1d , wherein R ^1a and R ^1d are each as defined herein. In certain embodiments, R ⁵ is -NR ^1a S(O) ₂ R ^1d , wherein R ^1a and R ^1d are each as defined herein. In certain embodiments, R ⁵ is -NR ^1a S(O)NR ^1b R ^1c , wherein each of R ^1a , R ^1b and R ^1c is as defined herein. In certain embodiments, R ⁵ is -NR ^1a S(O) ₂ NR ^1b R ^1c , wherein each of R ^1a , R ^1b and R ^1c is as defined herein. In certain embodiments, R ⁵ is -SR ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -S(O)R ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -S(O) ₂ R ^1a , wherein R ^1a is as defined herein. In certain embodiments, R ⁵ is -S(O)NR ^1b R ^1c , wherein R ^1b and R ^1c are each as defined herein. In certain embodiments, R ⁵ is -S(O) ₂ NR ^1b R ^1c , wherein R ^1b and R ^1c are each as defined herein.

In certain embodiments, the compound provided herein is not 1-(5-chloro-2,4-dihydroxybenzoyl)-N'-methyl-N'-(6-methylpyridin-3-ylmethyl base)-D-prolinamide.

In one embodiment, in the general formula provided herein, R ¹ is hydrogen or C _1-6 alkyl, optionally substituted with one or more substituents Q; R ² is heteroaryl or heteroaryl-C _1-6 alkyl, each optionally substituted by one or more substituents Q; and R ⁵ is halogen, C _1-6 alkyl, or C _3-7 cycloalkyl, wherein each of alkyl and cycloalkyl is optionally Substituted by one or more substituents Q.

In another embodiment, in the general formula provided herein, R ¹ is hydrogen or C _1-6 alkyl, optionally substituted with one or more substituents Q; R ² is 6-membered heteroaryl-C _1-6 alkyl, 5,6-fused bicyclic heteroaryl or 6,6-fused bicyclic heteroaryl, each optionally substituted by one or more substituents Q; and ^R is halogen, C _1-6 alkyl or C _3-7 cycloalkyl, wherein each of the alkyl and cycloalkyl is optionally substituted by one or more substituents Q.

In yet another embodiment, in the general formulas provided herein, ^R is hydrogen, methyl, ethyl, or propyl; ^R is pyridylmethyl, chloropyrazolo[1,5-a]pyrimidine base) methyl or quinolinylmethyl; and ^R is chloro, methyl or cyclopropyl.

In yet another embodiment, in the general formulas provided herein, R ¹ is hydrogen, methyl, ethyl, or isopropyl; R ² is pyridylmethyl, 6-chloropyrazolo[1,5- a] pyrimidin-3-yl)methyl or quinolinylmethyl; and R ⁵ is chloro, methyl or cyclopropyl.

及其药学可接受的盐、溶剂合物和水合物。在一个实施方案中，本文还提供下式化合物

及其药学可接受的盐、溶剂合物和水合物。药物组合物In one embodiment, the compounds provided herein are selected from:

and pharmaceutically acceptable salts, solvates and hydrates thereof. In one embodiment, also provided herein are compounds of the formula

and pharmaceutically acceptable salts, solvates and hydrates thereof. pharmaceutical composition

The pharmaceutical composition provided herein comprises the compound provided herein as an active ingredient, such as a compound of formula I, including an enantiomer, a mixture of enantiomers, or a mixture of diastereomers; or a pharmaceutically acceptable an acceptable salt, solvate, or hydrate; and a pharmaceutically acceptable medium, carrier, diluent, or excipient, or a mixture thereof.

Suitable excipients are well known to those skilled in the art, and non-limiting examples of suitable excipients are provided herein. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on a variety of factors well known to those skilled in the art, including, but not limited to, the method of administration. For example, oral dosage forms such as tablets may contain excipients not suitable for parenteral dosage forms. The suitability of a particular excipient also depends on the particular active ingredient in the dosage form. For example, the decomposition of some active ingredients may be accelerated by certain excipients such as lactose or when in contact with water. Active ingredients containing primary and secondary amines are particularly susceptible to this accelerated decomposition. Accordingly, the pharmaceutical compositions or dosage forms provided herein contain little, if any, lactose other mono- or di-saccharides. As used herein, the term "lactose-free" means that lactose, if any, is present in an amount substantially insufficient to increase the rate of degradation of the active ingredient. In one embodiment, a lactose-free composition comprises an active ingredient provided herein, a binder/filler, and a lubricant. In another embodiment, a lactose-free dosage form comprises an active ingredient, microcrystalline cellulose, pregelatinized starch, and magnesium stearate.

A compound provided herein can be administered alone or in combination with one or more other compounds provided herein. The pharmaceutical composition comprises a compound provided herein, such as a compound of formula I, including an enantiomer, a mixture of enantiomers, or a mixture of diastereomers; or a pharmaceutically acceptable salt or solvate thereof Composition, or hydrate, the pharmaceutical composition can be formulated into different dosage forms suitable for oral, parenteral and topical administration. The pharmaceutical composition can also be formulated in modified release dosage forms, including delayed-, extended-, extended-, sustained-, pulsed-, controlled-, accelerated-, rapid-, targeted-, pressed Programmed-release, and gastric retention dosage forms. These dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art (see, Remington: Science and Practice of Pharmacy, 21st ed.; Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Modified-Release Drug Delivery Technology, 2nd ed. .; Rathbone et al., Eds.; Marcel Dekker, Inc.: New York, NY, 2008).

In one embodiment, the pharmaceutical composition is provided in a dosage form for oral administration comprising a compound provided herein, such as a compound of Formula I, including an enantiomer, a mixture of enantiomers, or a diastereomer A mixture of enantiomers; or a pharmaceutically acceptable salt, solvate, or hydrate thereof; and one or more pharmaceutically acceptable excipients or carriers.

In another embodiment, the pharmaceutical composition is provided in a dosage form for parenteral administration comprising a compound provided herein, such as a compound of Formula I, including enantiomers, mixtures of enantiomers, or a mixture of diastereoisomers; or a pharmaceutically acceptable salt, solvate, or hydrate thereof; and one or more pharmaceutically acceptable excipients or carriers.

In yet another embodiment, the pharmaceutical composition is provided in a dosage form for topical administration comprising a compound provided herein, such as a compound of Formula I, including an enantiomer, a mixture of enantiomers, or a mixture of diastereoisomers; or a pharmaceutically acceptable salt, solvate, or hydrate thereof; and one or more pharmaceutically acceptable excipients or carriers.

The pharmaceutical compositions provided herein can be provided in single dose form or in multiple dose form. As used herein, unit-dose form refers to physically discrete units suitable for administration to human and animal subjects and packaged individually, as is known in the art. Each unit dosage contains a predetermined quantity of active ingredient sufficient to produce the desired therapeutic effect, in association with the required pharmaceutical carrier or excipient. Examples of unit-dose forms include ampoules, syringes, and individually packaged tablets and capsules. For example, a 100 mg unit dose will contain about 100 mg of the active ingredient in a packaged tablet or capsule. Single dose forms can be administered in fractions or multiples thereof. A multiple-dose form is a plurality of the same unit-dose form packaged in a single container, administered as separate unit-dose forms. Examples of multiple dose forms include glass bottles, tablet or capsule bottles, or pint or gallon bottles.

The pharmaceutical compositions provided herein can be administered once, or multiple times at regular intervals. It is understood that the precise dosage and duration of treatment may vary with the age, weight and condition of the patient being treated, and can be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test or diagnostic data. It is also understood that for any particular individual, the specific dosage regimen will be adjusted over time according to the needs of that individual and the professional judgment of the person administering or directing the administration of the formulation. Instructions

In one embodiment, provided herein is a method of treating or preventing a proliferative disease in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, such as a compound of formula I, including a single enantiomer isomer, a mixture of enantiomers, or a mixture of diastereomers; or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In another embodiment, provided herein is a method of treating, preventing or ameliorating one or more symptoms of a proliferative disease in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound provided herein, e.g. A compound of formula I, including a single enantiomer, a mixture of enantiomers, or a mixture of diastereomers; or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In yet another embodiment, provided herein is a method of treating, preventing, or ameliorating one or more symptoms of an HSP90-mediated disorder, disease, or condition in a subject, the method comprising administering to the subject a therapeutically effective Amounts of a compound provided herein, such as a compound of formula I, including a single enantiomer, a mixture of enantiomers, or a mixture of diastereomers; or a pharmaceutically acceptable salt, solvate, or Hydrate. In one embodiment, the HSP90-mediated disorder, disease or condition is a proliferative disease.

In certain embodiments, the subject is a mammal. In certain embodiments, the subject is a human.

In one embodiment, the proliferative disease is a tumor. In another embodiment, the proliferative disease is a solid tumor. In another embodiment, the proliferative disease is cancer. In certain embodiments, the proliferative disease is drug resistant cancer. In certain embodiments, the proliferative disease is metastatic cancer.

In certain embodiments, cancers that may be treated with the methods provided herein include, but are not limited to, bladder cancer, breast cancer, cervical cancer, colon cancer (e.g., colorectal cancer), esophageal cancer, head and neck cancer, liver cancer, lung cancer ( For example, small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, sarcoma (such as osteosarcoma), skin cancer (such as squamous cell cancer), gastric, testicular, thyroid and uterine cancers.

In one embodiment, provided herein is a method of inhibiting the growth of a cell comprising exposing the cell to a compound provided herein, such as a compound of Formula I, including a single enantiomer, a mixture of enantiomers, or a mixture of diastereoisomers; or a pharmaceutically acceptable salt, solvate or hydrate thereof.

In certain embodiments, the cell is a mammalian cell. In certain embodiments, the cells are human cells. In certain embodiments, the cells are tumor cells. In certain embodiments, the cell is a mammalian tumor cell. In certain embodiments, the cells are human tumor cells. In certain embodiments, the cell is a cancer cell. In certain embodiments, the cell is a mammalian cancer cell. In certain embodiments, the cell is a human cancer cell.

In certain embodiments, cancer cells that can be treated with the methods provided herein include, but are not limited to, bladder cancer, breast cancer, cervical cancer, colon cancer (e.g., colorectal cancer), esophageal cancer, head and neck cancer, liver cancer, lung cancer (e.g., small cell lung cancer and non-small cell lung cancer), melanoma, myeloma, neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, sarcomas (such as osteosarcoma), skin cancer (such as squamous cell carcinoma ), gastric, testicular, thyroid and uterine cancer cells.

In one embodiment, the cell is bladder cancer, squamous cell carcinoma, head and neck cancer, colorectal cancer, esophageal cancer, gastric cancer, gynecological cancer, pancreatic cancer, rectal cancer, breast cancer, prostate cancer, vulvar cancer, skin cancer, Cells from cancers of the brain, genitourinary tract, lymphatic system, stomach, larynx, or lung.

In certain embodiments, the cancer cell is a metastatic cancer cell, including but not limited to bladder cancer, breast cancer, cervical cancer, colon cancer (e.g., colorectal cancer), esophageal cancer, head and neck cancer, liver cancer, lung cancer (e.g., small Lung cancer and non-small cell lung cancer), melanoma, myeloma, neuroblastoma, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, sarcomas (such as osteosarcoma), skin cancer (such as squamous cell carcinoma) , gastric, testicular, thyroid and uterine cancer cells.

Inhibition of cell growth can be quantified, for example, by counting the number of cells exposed to the compound of interest compared to otherwise identical cells not contacted with the compound, or by measuring the number of cells containing the cell. the size of the tumor. Cell number as well as cell size can be readily assessed using any method known in the art (e.g., trypan blue exclusion and cell counting, measuring ³ H- thymidine).

In one embodiment, provided herein is a method for inhibiting HSP90 enzyme activity, the method comprising making HSP90 enzyme and a compound of formula I provided herein, including a single enantiomer, a mixture of enantiomers, or a diastereomer A mixture of enantiomers; or a pharmaceutically acceptable salt, solvate or hydrate thereof.

The present disclosure will be further understood by the following non-limiting examples.

Detailed ways

Example

2，4-二(甲氧基甲氧基)苯甲酸甲酯是如下使用一般程序制备的：在N₂气氛下，将氯甲基甲基醚(26mL，342mmol)和DIEA(84ml_，480mmol)加至2，4-二羟基苯甲酸甲酯(13.4g，80mmol)在40mL的DMF中的反应溶液中。将所得混合物在室温下搅拌12小时。使反应混合物在EtOAc(3x 500mL)与水(200mL)、饱和NaHCO₃溶液(2x 200mL)和盐水(200mL)之间分配。使有机层干燥(Na₂SO₄)，过滤，再通过真空浓缩，得到油状残余物。将该残余物通过硅胶色谱(梯度洗脱液0→30％EtOAc/己烷)纯化，得到需要的中间体产物2，4-二(甲氧基甲氧基)苯甲酸甲酯(19.20g，93％产率)。¹H NMR(400MHz，氯仿-D)δppm 3.47(s，3H)3.51(s，3H)3.85(s，3H)5.18(s，2H)5.23(s，2H)6.70(dd，J＝8.84，2.27Hz，1H)6.83(d，J＝2.27Hz，1H)7.80(d，J＝8.84Hz，1H)。然后如下制备5-氯-2，4-二(甲氧基甲氧基)苯甲酸甲酯。在10％HOAc中的0.7M次氯酸钙溶液的制备：在冰浴中搅拌下将次氯酸钙(9.743g，44.29mmol)溶解于64mL的10％HOAc中。将次氯酸钙溶液滴加至2，4-二(甲氧基甲氧基)苯甲酸甲酯(如上文制备)(9.45g，36.9mmol)在50.0mL丙酮中的反应溶液中。将所得混合物在室温下搅拌12小时。使反应混合物在EtOAc(3x 500mL)与饱和NaHCO₃溶液(200mL)之间分配。使有机层干燥(Na₂SO₄)，过滤，然后通过真空浓缩。将残余物通过硅胶色谱纯化(梯度洗脱液0→30％EtOAc/己烷)得到需要的产物5-氯-2，4-二(甲氧基甲氧基)苯甲酸甲酯(6.6g，62％产率)。¹H NMR(400MHz，氯仿-D)δppm(400MHz，氯仿-D)3.51(s，6H)3.86(s，3H)，5.22(s，2H)5.27(s，2H)7.03(s，1H)7.88(s，1H)。5-氯-2，4-二(甲氧基甲氧基)苯甲酸：然后将LiOH水溶液(40mL，2.0M)加至5-氯-2，4-二(甲氧基甲氧基)苯甲酸甲酯(5.8g，20mmol_在THF∶H₂O(4∶1，80mL)中的反应溶液中。将所得混合物在室温下搅拌12小时。将反应混合物通过真空浓缩，除去大部分溶剂，然后用醚(200mL)萃取。使用2.0N HCl溶液将水层中和至pH 6.5，然后萃取CH₂Cl₂(5x 100mL)。将合并的有机层干燥(Na₂SO₄)，然后通过真空浓缩，得到5.2g的需要的最终产物，5-氯-2，4-二(甲氧基甲氧基)苯甲酸，为白色固体(92％产率)。¹H NMR(400MHz，DMSO-D6)δppm¹H NMR(400MHz，DMSO-D6)δppm 3.52-3.56(d，J＝8.34Hz，6H)5.29(s，2H)5.39(s，2H)7.10(s，1H)，8.17(s，1H)。

5-环丙基-2，4-二(甲氧基甲氧基)苯甲酸甲酯：在氮气下向火焰干燥的烧瓶中加入5-氯-2，4-二(甲氧基甲氧基)苯甲酸甲酯(3.625g，12.5mmol)和环丙基硼酸(0.86g，10.0mmol)，接着加入在无水甲苯(250mL)中的碘化钠(186mg，1.25mmol)、K₂CO₃(2.76g，20.0mmol)、18-冠-6(2.64g，10.0mmol)、Pd(OAc)₂45.00mg(0.2mmol，2mol％)和二氢咪唑鎓(dihydroimidazolium)盐酸盐配体85mg(0.2mmol，2mol％)。将所得混悬液在80℃下搅拌12小时。使该反应混合物浓缩，然后加入二氯甲烷(10mL)。然后将该二氯甲烷溶液用NaHCO₃水溶液、盐水溶液洗涤3次，再用无水硫酸镁干燥。通过旋转蒸发浓缩溶剂，再将该粗物质通过硅胶色谱使用乙酸乙酯/己烷(5-30％)纯化。得到1.57克白色固体(53％产率)。¹H NMR(400MHz，氯仿-D)δppm1.22-1.24(m，4H)，1.52(m，1H)，3.51(s，6H)3.86(s，3H)，5.22(s，2H)5.27(s，2H)7.03(s，1H)7.88(s，1H)。5-环丙基-2，4-二(甲氧基甲氧基)苯甲酸：然后将LiOH水溶液(10mL，2.0M)加至5-环丙基-2，4-二(甲氧基甲氧基)苯甲酸甲酯(1.5g，5.0mmol)在THF∶H₂O(4∶1，20mL)中的反应溶液中。将所得混合物在室温下搅拌12小时。将反应混合物通过真空浓缩，除去大部分溶剂，然后用醚(100mL)萃取。使用2.0N HCl溶液将水层中和至pH 6.5，然后萃取CH₂Cl₂(5x 100mL)。将合并的有机层干燥(Na₂SO₄)，然后通过真空浓缩，得到1.27g的需要的最终产物，5-环丙基-2，4-二(甲氧基甲氧基)苯甲酸，为白色固体(90％产率)。¹H NMR(400MHz，DMSO-D6)δppm 1.21-1.24(m，4H)，1.51(m，1H)，3.52-3.56(d，J＝8.34Hz，6H)5.29(s，2H)5.39(s，2H)7.10(s，1H)，8.17(s，1H)。

S-2-(异丙基(吡啶-3-基甲基)氨基甲酰基)吡咯烷-1-甲酸酯：在氩气氛下将N-异丙基-N-(3-吡啶基甲基)胺(1.95g，13.0mmol)加至Boc-L-Pro-OH(3.655g，17.0mmol)、DIEA(500.0mmol)和HATU(110mmol)在150.0mL的DMF中的溶液中。使反应在室温下搅拌12小时，然后加入饱和NaHCO₃以猝灭反应。通过使用EtOAc萃取反应物。合并EtOAc层，用Na₂SO₄干燥。浓缩有机部分，再通过硅胶色谱用EtOAc/己烷纯化，得到产物3.88g(86％产率)。¹H NMR(400MHz，DMSO-D6)δppm：1.08(m，6H)，1.30(m，3H)，1.41-1.44(m，6H)，1.84-1.86(m，2H)，1.92-2.35(m，2H)，2.62-2.84(m，2H)，3.48-3.61(m，2H)，4.59-4.67(m，2H)，7.67-7.71(m，1H)，7.99(s，1H)，8.48-8.52(m，2H)。S-N-异丙基-N-(吡啶-3-基甲基)吡咯烷-2-甲酰胺：将氯化氢(8ml，32mmol，4.0M，在二氧杂环己烷中)加至以上化合物(660mg，1.9mmol)在DCM 5.0mL中的溶液中。将反应在室温下搅拌12小时。将该混合物浓缩，得到胶状产物，其未经纯化即用于下一步骤。(S)-1-(5-环丙基-2，4-二(甲氧基甲氧基)苯甲酰基)-N-异丙基-N-(吡啶-2-基甲基)吡咯烷-2-甲酰胺：在氩气氛下将S-N-异丙基-N-(吡啶-3-基甲基)吡咯烷-2-甲酰胺(321mg，1.3mmol)加至5-环丙基-2，4-二(甲氧基甲氧基)苯甲酸(423mg，1.5mmol)、DIEA(5.0mmol)和HATU(1.1mmol)在10.0mL的DMF中的溶液中。使反应在室温下搅拌12小时，然后加入饱和NaHCO₃以猝灭反应。通过使用EtOAc萃取反应物三次。合并EtOAc层，用Na₂SO₄干燥。浓缩有机部分，再通过硅胶色谱用EtOAc/己烷(20％至50％)纯化，得到黄-白色固体产物479mg(72％产率)。(S)-1-(5-环丙基-2，4-二羟基苯甲酰基)-N-异丙基-N-(吡啶-2-基甲基)吡咯烷-2-甲酰胺：将氯化氢(2ml，8mmol，4.0M，在二氧杂环己烷中)加至以上化合物(185mg，0.36mmol)在DCM 5.0mL中的溶液中。将反应在室温下搅拌12小时。将该混合物用饱和NaHCO₃(水溶液)中和，然后用EtOAc(3x20mL)萃取。合并有机部分，再在EtOAc/己烷(20％至60％)在硅胶色谱中浓缩以纯化，得到127mg(82％产率)；¹HNMR(400MHz，DMSO-D6)δppm 0.53(m，2H)，0.78(m，2H)，1.18(m，6H)，1.84-1.86(m，2H)，1.92-2.35(m，3H)，2.63-2.84(m，2H)，3.49-3.60(m，2H)，4.33-4.75(m，2H)，6.40(s，1H)，6.72(s，1H)，7.67-7.71(m，1H)，8.02(m，1H)，8.48-8.52(m，2H)。参考以上操作方法，本领域技术人员结合已有知识可以容易地合成以下各化合物。

生物活性Hsp90细胞分析使用Akt luminex分析法对基于细胞的抗HSP-90的效价进行评价，该分析法测定HSP-90客户蛋白Akt的翻转。将NCI-H1299细胞用HSP-90化合物的系列稀释液处理24小时。然后通过Akt/PKBBeadmate使用luminex 100系统，分析细胞溶胞产物，以测定Akt表达的损失，由此确定细胞IC₅₀。发现本发明化合物1、3、4、5、11、12、13、15、16抑制NCI-H1299细胞系具有小于1.0微摩尔的IC₅₀值。抗增生活性：本发明化合物的抗增生活性是在一定数量的细胞系例如人结肠癌细胞系HCT116中抑制细胞生长的能力来测定的。细胞生长的抑制作用是使用Alamar Blue分析法测定的。该方法是以存活细胞使刃天青(resazurin)还原成其荧光产物试卤灵(resorufin)的能力为基础的。对于每一增生分析，使细胞辅板到96孔板上，再使之恢复16小时，然后添加抑制剂化合物达另外6小时，然后在535nM ex/590nM em处测定荧光产物。在非增生分析的情况下，使细胞保持融合达96小时，然后添加抑制剂化合物达另外72小时。如前所述通过Alamar Blue分析法测定存活细胞的数量。发现本发明化合物1、2、3、4、5、6、7、8、9、10、11、13、15、16抑制HCT细胞系具有小于1.0微摩尔的IC₅₀值。化合物12和14抑制HCT细胞系具有小于5.0微摩尔的IC₅₀值。＊＊＊＊＊For all of the following examples, standard manipulations and purification methods known to those skilled in the art can be used. All temperatures are in °C (degrees Celsius) unless otherwise indicated. All reactions were performed at room temperature unless otherwise indicated. The synthetic methods described in Schemes 4 to 18 are intended to illustrate the available chemistry by using specific examples and are not indicative of the scope of the disclosure. Example 1

Methyl 2,4-bis(methoxymethoxy)benzoate was prepared using the general procedure as follows: chloromethyl methyl ether (26 mL, 342 mmol) and DIEA (84 mL, 480 mmol) were mixed under _N atmosphere Add to a reaction solution of methyl 2,4-dihydroxybenzoate (13.4 g, 80 mmol) in 40 mL of DMF. The resulting mixture was stirred at room temperature for 12 hours. The reaction mixture was partitioned between EtOAc (3 x 500 mL) and water (200 mL), saturated NaHCO ₃ solution (2 x 200 mL) and brine (200 mL). The organic layer was dried ( _Na2SO4 ), filtered, and concentrated by vacuum _to give an oily residue. The residue was purified by silica gel chromatography (gradient eluent 0→30% EtOAc/hexanes) to give the desired intermediate product methyl 2,4-bis(methoxymethoxy)benzoate (19.20 g, 93% yield). ¹ H NMR (400MHz, chloroform-D) δppm 3.47(s, 3H) 3.51(s, 3H) 3.85(s, 3H) 5.18(s, 2H) 5.23(s, 2H) 6.70(dd, J=8.84, 2.27 Hz, 1H) 6.83 (d, J=2.27Hz, 1H) 7.80 (d, J=8.84Hz, 1H). Methyl 5-chloro-2,4-bis(methoxymethoxy)benzoate was then prepared as follows. Preparation of 0.7M calcium hypochlorite solution in 10% HOAc: Calcium hypochlorite (9.743 g, 44.29 mmol) was dissolved in 64 mL of 10% HOAc with stirring in an ice bath. The calcium hypochlorite solution was added dropwise to a reaction solution of methyl 2,4-bis(methoxymethoxy)benzoate (prepared as above) (9.45 g, 36.9 mmol) in 50.0 mL of acetone. The resulting mixture was stirred at room temperature for 12 hours. The reaction mixture was partitioned between EtOAc (3 x 500 mL) and saturated NaHCO ₃ solution (200 mL). The organic layer was dried (Na ₂ SO ₄ ), filtered, and concentrated by vacuum. The residue was purified by silica gel chromatography (gradient eluent 0→30% EtOAc/hexanes) to give the desired product, methyl 5-chloro-2,4-bis(methoxymethoxy)benzoate (6.6 g, 62% yield). ¹ H NMR (400MHz, chloroform-D) δppm (400MHz, chloroform-D) 3.51 (s, 6H) 3.86 (s, 3H), 5.22 (s, 2H) 5.27 (s, 2H) 7.03 (s, 1H) 7.88 (s, 1H). 5-Chloro-2,4-bis(methoxymethoxy)benzoic acid: Then aqueous LiOH (40 mL, 2.0 M) was added to 5-chloro-2,4-bis(methoxymethoxy)benzene Methyl formate (5.8 g, 20 mmol) in a reaction solution in THF: _H2O (4:1, 80 mL). The resulting mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated by vacuum to remove most of the solvent, Then extracted with ether (200 mL). The aqueous layer was neutralized to pH 6.5 using 2.0 N HCl solution, then CH ₂ Cl ₂ (5 x 100 mL) was extracted. The combined organic layers were dried (Na ₂ SO ₄ ) and concentrated by vacuum , to obtain 5.2 g of the desired final product, 5-chloro-2,4-bis(methoxymethoxy)benzoic acid, as a white solid (92% yield) ^.1H NMR (400MHz, DMSO-D6) δppm ¹ H NMR (400MHz, DMSO-D6) δppm 3.52-3.56(d, J=8.34Hz, 6H) 5.29(s, 2H) 5.39(s, 2H) 7.10(s, 1H), 8.17(s, 1H) .

Methyl 5-cyclopropyl-2,4-bis(methoxymethoxy)benzoate: To a flame-dried flask under nitrogen was added 5-chloro-2,4-bis(methoxymethoxy ) methyl benzoate (3.625 g, 12.5 mmol) and cyclopropylboronic acid (0.86 g, 10.0 mmol), followed by addition of sodium iodide (186 mg, 1.25 mmol), K ₂ CO ₃ in anhydrous toluene (250 mL) (2.76g, 20.0mmol), 18-crown-6 (2.64g, 10.0mmol), Pd(OAc) ₂ 45.00mg (0.2mmol, 2mol%) and dihydroimidazolium (dihydroimidazolium) hydrochloride ligand 85mg ( 0.2 mmol, 2 mol%). The resulting suspension was stirred at 80°C for 12 hours. The reaction mixture was concentrated, then dichloromethane (10 mL) was added. The dichloromethane solution was then washed three times with NaHCO ₃ aqueous solution and brine solution, and dried over anhydrous magnesium sulfate. The solvent was concentrated by rotary evaporation and the crude material was purified by silica gel chromatography using ethyl acetate/hexane (5-30%). Obtained 1.57 g of white solid (53% yield). ¹ H NMR (400MHz, chloroform-D) δppm1.22-1.24 (m, 4H), 1.52 (m, 1H), 3.51 (s, 6H) 3.86 (s, 3H), 5.22 (s, 2H) 5.27 (s , 2H) 7.03(s, 1H) 7.88(s, 1H). 5-Cyclopropyl-2,4-bis(methoxymethoxy)benzoic acid: Aqueous LiOH (10 mL, 2.0 M) was then added to 5-cyclopropyl-2,4-bis(methoxymethyl oxy)methylbenzoate (1.5 g, 5.0 mmol) in THF:H ₂ O (4:1, 20 mL) in the reaction solution. The resulting mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated by vacuum to remove most of the solvent, then extracted with ether (100 mL). The aqueous layer was neutralized to pH 6.5 using 2.0N HCl solution, then _{CH2Cl2 was} extracted (5 x 100 mL). The combined organic layers were dried (Na ₂ SO ₄ ) and then concentrated by vacuum to give 1.27 g of the desired final product, 5-cyclopropyl-2,4-bis(methoxymethoxy)benzoic acid, as White solid (90% yield). ¹ H NMR (400MHz, DMSO-D6) δppm 1.21-1.24(m, 4H), 1.51(m, 1H), 3.52-3.56(d, J=8.34Hz, 6H) 5.29(s, 2H) 5.39(s, 2H) 7.10(s, 1H), 8.17(s, 1H).

S-2-(isopropyl(pyridin-3-ylmethyl)carbamoyl)pyrrolidine-1-carboxylate: N-isopropyl-N-(3-pyridylmethyl ) amine (1.95 g, 13.0 mmol) was added to a solution of Boc-L-Pro-OH (3.655 g, 17.0 mmol), DIEA (500.0 mmol) and HATU (110 mmol) in 150.0 mL of DMF. The reaction was allowed to stir at room temperature for 12 hours, then was quenched by the addition of saturated NaHCO ₃ . The reaction was extracted by using EtOAc. The EtOAc layers were combined and dried _{over Na2SO4} . The organic portion was concentrated and purified by silica gel chromatography with EtOAc/hexanes to give the product 3.88g (86% yield). ¹ H NMR (400MHz, DMSO-D6) δppm: 1.08(m, 6H), 1.30(m, 3H), 1.41-1.44(m, 6H), 1.84-1.86(m, 2H), 1.92-2.35(m, 2H), 2.62-2.84(m, 2H), 3.48-3.61(m, 2H), 4.59-4.67(m, 2H), 7.67-7.71(m, 1H), 7.99(s, 1H), 8.48-8.52( m, 2H). SN-Isopropyl-N-(pyridin-3-ylmethyl)pyrrolidine-2-carboxamide: Add hydrogen chloride (8ml, 32mmol, 4.0M in dioxane) to the above compound (660mg , 1.9 mmol) in a solution in DCM 5.0 mL. The reaction was stirred at room temperature for 12 hours. The mixture was concentrated to give a gum product which was used in the next step without purification. (S)-1-(5-cyclopropyl-2,4-bis(methoxymethoxy)benzoyl)-N-isopropyl-N-(pyridin-2-ylmethyl)pyrrolidine -2-Carboxamide: Add SN-isopropyl-N-(pyridin-3-ylmethyl)pyrrolidine-2-carboxamide (321 mg, 1.3 mmol) to 5-cyclopropyl-2 under argon atmosphere , in a solution of 4-bis(methoxymethoxy)benzoic acid (423 mg, 1.5 mmol), DIEA (5.0 mmol) and HATU (1.1 mmol) in 10.0 mL of DMF. The reaction was allowed to stir at room temperature for 12 hours, then was quenched by the addition of saturated NaHCO ₃ . The reaction was extracted three times by using EtOAc. The EtOAc layers were combined and dried _{over Na2SO4} . The organic portion was concentrated and purified by silica gel chromatography with EtOAc/hexanes (20% to 50%) to give the product as a yellow-white solid 479 mg (72% yield). (S)-1-(5-cyclopropyl-2,4-dihydroxybenzoyl)-N-isopropyl-N-(pyridin-2-ylmethyl)pyrrolidine-2-carboxamide: the Hydrogen chloride (2 ml, 8 mmol, 4.0 M in dioxane) was added to a solution of the above compound (185 mg, 0.36 mmol) in DCM 5.0 mL. The reaction was stirred at room temperature for 12 hours. The mixture was neutralized with saturated NaHCO ₃ (aq), then extracted with EtOAc (3×20 mL). The combined organic fractions were concentrated and purified by chromatography on silica gel in EtOAc/hexanes (20% to 60%) to give 127 mg (82% yield); ¹ HNMR (400 MHz, DMSO-D6) δ ppm 0.53 (m, 2H) , 0.78(m, 2H), 1.18(m, 6H), 1.84-1.86(m, 2H), 1.92-2.35(m, 3H), 2.63-2.84(m, 2H), 3.49-3.60(m, 2H) , 4.33-4.75 (m, 2H), 6.40 (s, 1H), 6.72 (s, 1H), 7.67-7.71 (m, 1H), 8.02 (m, 1H), 8.48-8.52 (m, 2H). Referring to the above operation methods, those skilled in the art can easily synthesize the following compounds in combination with existing knowledge.

Bioactive Hsp90 Cellular Assay Cell-based anti-HSP-90 potency was evaluated using the Akt luminex assay, which measures turnover of the HSP-90 client protein Akt. NCI-H1299 cells were treated with serial dilutions of HSP-90 compounds for 24 hours. Cell lysates were then analyzed by Akt/PKB Beadmate using the luminex 100 system to determine the loss of Akt expression, thereby determining the cellular _IC50 . Compounds 1, 3, 4, 5, 11, 12, 13, 15, 16 of the invention were found to inhibit the NCI-H1299 cell line with _IC50 values less than 1.0 micromolar. Antiproliferative activity: The antiproliferative activity of the compounds of the present invention is determined by the ability to inhibit cell growth in a number of cell lines, such as the human colon carcinoma cell line HCT116. Inhibition of cell growth was determined using the Alamar Blue assay. The method is based on the ability of surviving cells to reduce resazurin to its fluorescent product resorufin. For each proliferation assay, cells were plated onto 96-well plates, allowed to recover for an additional 16 hours, and then inhibitor compounds were added for an additional 6 hours before fluorescence product was measured at 535nM ex/590nM em. In the case of non-proliferative assays, cells were kept confluent for 96 hours before addition of inhibitor compounds for an additional 72 hours. The number of surviving cells was determined by Alamar Blue assay as previously described. Compounds 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 13, 15, 16 of the invention were found to inhibit HCT cell lines with _IC50 values less than 1.0 micromolar. Compounds 12 and 14 inhibited HCT cell lines with _IC50 values of less than 5.0 micromolar. ＊＊＊＊＊

The above examples are provided to those skilled in the art to fully disclose and describe how to make and use the claimed embodiments and not to limit the scope of the disclosure herein. Modifications obvious to those skilled in the art are intended to be within the scope of the appended claims. All publications, patents, and patent applications cited in this specification are herein incorporated by reference as if each such publication, patent, and patent application were specifically and individually indicated to be incorporated by reference.

Claims (10)

1. Formula I compound: or a single enantiomer, a mixture of enantiomers or a mixture of diastereomers; or a pharmaceutically acceptable salt, solvate or hydrate thereof; in: R ¹ is hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 aryl, C _7-15 aralkyl, hetero Aryl or heterocyclic; R ² is heteroaryl or heteroaryl-C _1-6 alkyl; R ⁵ is (i) hydrogen, halogen, cyano or nitro; (ii) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _{6 -14} aryl, C _7-15 aralkyl, heteroaryl or heterocyclyl; or (iii) -C(O)R ^1a , -C(O)OR ^1a , -C(O)NR ^1b R ^1c , -C(NR ^1a )NR ^1b R ^1c , -OR ^1a , -OC(O)R ^1a , -OC(O)OR ^1a , -OC(O)NR ^1b R ^1c , -OC(=NR ^1a )NR ^1b R ^1c , -OS(O)R ^1a , -OS(O) ₂ R ^1a , -OS(O)NR ^1b R ^1c , -OS(O) ₂ NR ^1b R ^1c , -NR ^1b R ^1c , -NR ^1a C(O)R ^1d , -NR ^1a C(O)OR ^1d , -NR ^1a C(O)NR ^1b R ^1c , -NR ^1a C(=NR ^1d )NR ^1b R ^1c , -NR ^1a S(O )R ^1d , -NR ^1a S(O) ₂ R ^1d , -NR ^1a S(O)NR ^1b R ^1c , -NR ^1a S(O) ₂ NR ^1b R ^1c , -SR ^1a , -S(O)R ^1a , -S(O) _2R1a ^, -S(O ^{) NR1bR1c ,} or _-S (O) ^2NR1bR1c ; and Each of R ^1a , R ^1b , R ^1c and R ^1d is independently hydrogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 Aryl, C _7-15 aralkyl, heteroaryl or heterocyclic group; or R ^1b and R ^1c form heteroaryl or heterocyclic group together with their connected N atom; Provided that the compound is not 1-(5-chloro-2,4-dihydroxybenzoyl)-N'-methyl-N'-(6-methylpyridin-3-ylmethyl)-D-proline amides; wherein each alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl and heteroaryl-alkyl is optionally substituted by one or more groups Q, each Q independently selected from (a) cyano, halogen and nitro; (b) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _{6- Aryl} , C _7-15 aralkyl, heteroaryl and heterocyclyl, each optionally substituted by one or more (in one embodiment, 1, 2, 3 or 4) substituents ^Q and (c)-C(O)R ^a , -C(O)OR ^a , -C(O)NR ^b R ^c , -C(NR ^a )NR ^b R ^c , -OR ^a , -OC(O )R ^a , -OC(O)OR ^a , -OC(O)NR ^b R ^c , -OC(=NR ^a )NR ^b R ^c , -OS(O)R ^a , -OS(O) ₂ R ^a , -OS(O)NR ^b R ^c , -OS(O) ₂ NR ^b R ^c , -NR ^b R ^c , -NR ^a C(O)R ^d , -NR ^a C(O)OR ^d , -NR ^a C(O)NR ^b R ^c , -NR ^a C(=NR ^d )NR ^b R ^c , -NR ^a S(O)R ^d , -NR ^a S(O) ₂ R ^d , -NR ^a S( O)NR ^b R ^c , -NR ^a S(O) ₂ NR ^b R ^c , -SR ^a , -S(O)R ^a , -S(O) ₂ R ^a , -S(O)NR ^b R ^c and -S(O) ₂ NR ^b R ^c , wherein each of R ^a , R ^b , R ^c and R ^d is independently (i) hydrogen; (ii) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 aryl, C _7-15 aralkyl, heteroaryl or heterocyclyl, each optionally replaced by one or more (in one implementation In the scheme, it is 1, 2, 3 or 4) substituent Q ^a substituted; or (iii) R ^b and R ^c form a heterocyclic group together with their connected N atoms, optionally by one or more (in one In one embodiment, 1, 2, 3 or 4) substituent ^{Qa is} substituted; Wherein each Q ^a is independently selected from (a) cyano, halogen and nitro; (b) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl , C _6-14 aryl, C _7-15 aralkyl, heteroaryl and heterocyclyl; and (c)-C(O)R ^e , -C(O)OR ^e , -C(O)NR ^f R ^g , -C(NR ^e )NR ^f R ^g , -OR ^e , -OC(O)R ^e , -OC(O)OR ^e , -OC(O)NR ^f R ^g , -OC(=NR ^e )NR ^f R ^g , -OS(O)R ^e , -OS(O) ₂ R ^e , -OS(O)NR ^f R ^g , -OS(O) ₂ NR ^f R ^g , -NR ^f R ^g , -NR ^e C(O)R ^h , -NR ^e C(O)OR ^f , -NR ^e C(O)NR ^f R ^g , -NR ^e C(=NR ^h )NR ^f R ^g , -NR ^e S(O)R ^h , -NR ^e S(O) ₂ R ^h , -NR ^e S(O)NR ^f R ^g , -NR ^e S(O) ₂ NR ^f R ^g , -SR ^e , -S( O ^{) Re} , -S(O ⁾ _2Re , -S(O) ^{NRfRg ,} and -S(O) _2NRfRg ^; wherein each ^Re , ^Rf , ^Rg , and ^{Rh are} independently is (i) hydrogen; (ii) C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-7 cycloalkyl, C _6-14 aryl, C _7-15 aryl Alkyl, heteroaryl or heterocyclyl; or (iii) ^Rf and ^Rg together with the N atom to which they are attached form a heterocyclyl. 2. The compound of claim 1, wherein the compound has the structure of formula II:

3. The compound of claim 1 or 2, wherein R ¹ is hydrogen or C _1-6 alkyl, optionally substituted by one or more substituents Q. 4. The compound of claim 3, wherein R ¹ is hydrogen, methyl, ethyl or propyl. 5. The compound of any one of claims 1 to 4, wherein R ² is heteroaryl-C _1-6 alkyl, each optionally substituted by one or more substituents Q. 6. The compound of claim 5, characterized in that: R ² is 6-membered heteroaryl-C _1-6 alkyl, 5,6-fused bicyclic heteroaryl-C _1-6 alkyl or 5,6-fused bicyclic heteroaryl-C _{1 -6} alkyl groups, each optionally substituted by one or more substituents Q; or R ² is pyridylmethyl, pyrazolo[1,5-a]pyrimidin-3-yl)methyl or quinolinyl-methyl, each optionally substituted by one or more substituents Q. 7. The compound of any one of claims 1 to 6, wherein R is halogen, C ^1-6 alkyl or C _3-7 cycloalkyl, wherein _each of the alkyl and cycloalkyl is optionally replaced by one or more The substituent Q is substituted. 8. The compound of claim 7, wherein R ⁵ is chloro, methyl or cyclopropyl. 9. The compound of claim 1 selected from the group consisting of:

and pharmaceutically acceptable salts, solvates and hydrates thereof. 10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 9, and a pharmaceutically acceptable excipient or carrier.