WO2017045612A1 - Pyrimidine derivative pim kinase inhibitor, preparation method therefor, and application thereof in medicine preparation - Google Patents

Abstract

A PIM kinase inhibitor, a preparation method therefor, and application thereof. A structural formula thereof is a compound as shown in a following general formula I. Also provided are a stereoisomer, a tautomer, and pharmaceutical salt of the compound in the general formula I. The compound of the general formula I has an obvious inhibiting function on the activity of kinase, can be used as the PIM kinase inhibitor, can be used for preparing medicine for treating or preventing cancer, reversing the drug resistance of anticancer medicine and other medicine, treating or preventing diseases of inflammatory bowel disease, autoimmune diseases and the like, and has a great clinical application value.

Description

Pyrimidine derivative PIM kinase inhibitor and preparation method thereof and application in pharmacy

cross reference

This application claims priority to Chinese Patent Application No. 201510594163.1, entitled "Pyridine Pyridine Derivative PIM Kinase Inhibitor, Preparation Method and Application in Pharmaceuticals", which is filed on September 18, 2015. The entire contents are incorporated herein by reference.

Technical field

The present invention relates to medicinal chemistry, and in particular to PIM kinase inhibitors, methods for their preparation and their use in pharmacy.

Background technique

PIM kinases are three homologous serine/threonine kinases belonging to the family of calmodulin-dependent protein kinases (CAMK). Studies have shown that PIM kinase is widely expressed in hematopoietic tissues (J. Biol. Chem., 280, 14168-14176, 2005; Blood, 105, 4477-4483, 2005) and plays an important role in cell survival and proliferation. And overexpressed in human cancer tumors as well as in inflammatory conditions (J. Exp. Med., 201, 259-266, 2005; Biochem. Soc. Trans., 32, 315-319, 2004). Therefore, PIM kinase is increasingly being used to study targets for the treatment of tumors and immunomodulatory drugs. The PIM-1 (Provirus Integration of Maloney 1) gene is a site for frequent insertion of provirus in Moloney murine leukemia virus-induced T cell lymphoma, and PIM-1 kinase is also named (Cell, 37, 141-150, 1984). It was later found that the gene encoding PIM-2 (Provirus Integration of Maloney 2) also had the same weakness (J. Clin. Invest., 115, 2679-2688, 2005). PIM-3 was originally named KID-1 (Kinase Induced by Depolarization 1) and was later renamed for its highly consistent protein sequence with PIM-1 (71% amino acid repeat) (Nature, 428, 332-337, 2005; Cell, 56, 673- 682, 1989). PIM-1, 2, 3 are overexpressed in many hematological tumors (PNAS USA, 86, 8857-8861, 1989). PIM-1 has been found to be more expressed during the development of prostate cancer (J. Clin. Pathol., 59, 285-288, 2006), and PIM-2 is found in human chronic lymphocytic leukemia and non-Hodgkin's lymphoma. Expression is increased (Leuk. Lymph., 45, 951-955, 2004), while abnormal expression of PIM-3 is considered for hepatic fibroma (Int. J. Cancer, 114, 209-218, 2005) and pancreatic cancer (Cancer Res) The development and diffusion of 66, 6741-6747, 2006) played an important role.

PIM-1, 2, 3 usually respond to stimulation by growth factors and cytokines, thus contributing to the survival and spread of hematopoietic cells. Mice that knocked out the PIM-1, 2, and 3 genes survived normally, but were smaller and spread during hematopoietic cell proliferation. The response to growth factors has also been reduced. If only one of the three PIMs was removed, there was no significant effect on the mice, and the functions of the three PIMs were overlapped (Cell, 56, 673-682, 1989). Substrates of action of PIM kinase include Bcl-2 family members that regulate apoptosis (FEBS Letters, 571, 43-49, 2004), p21 that regulates cell cycle (Biochim. Biophys. Acta, 1593, 45-55, 2002) ), CDC25A, C-TA, (J. Biol. Chem., 279, 48319-48328, 2004), and 4EBP1 (Blood, 105, 4477-4483, 2005) regulating protein synthesis. These effects of PIM kinase have been shown to have functions that prevent apoptosis and promote cell growth and spread. Therefore, overexpression of PIM kinase in tumors contributes to the survival and spread of cancer cells. Therefore, inhibition of overexpression of PIM kinase in tumors is a new and effective method for treating cancer.

Since PIM kinase is involved in many liquid and solid tumors, many PIM kinase inhibitors have been used to develop a new generation of anti-tumor drugs. In a series of cell and animal model experiments, PIM kinase inhibitors can significantly inhibit the proliferation of tumor cells including acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), and chronic tumor growth. Granulocyte leukemia (CML), non-Hodgkin's lymphoma (NHL), and multiple myeloma (MM) (Clin. Cancer Res. 20 (7), 1834–1845, 2014; Blood 123 (6), 905– 913, 2014; Blood 122 (21), 4435, 2013). The results also show that PIM kinase inhibitors also have a good effect on solid tumors overexpressing PIM kinase, including pancreatic cancer (Cancer Biol. Ther. 7 (9), 1352-9, 2008 Cancer Res. 2006; 66 (13 ): 6741-7; Cancer Res. 70 (24), 10288–10298, 2010), Prostate Cancer (Prostate, 65(3), 276-86, 2005; Prostate, 73(13), 1462–1469, 2013) , Liver cancer (J. Surg. Res., 153 (1), 17-22, 2009; Int. J. Cancer, 114 (2), 209-18, 2005), gastric cancer (J. Cancer Res. Clin. Oncol. , 134 (4), 481-8, 2008) and bladder cancer (J. Exp. Clin. Cancer Res., 29, 161, 2010).

The emergence of anticancer drug resistance has been a major obstacle to chemotherapy and molecular targeted drugs (Drug Resistance Updat.12, 114–126.2009), and addressing the drug resistance of anticancer drugs is an important topic in cancer therapy. Many studies have shown that PIM kinase is involved in the expression and activity of two of the most important drug efflux transporters, MDR-1 and BCRP (Drug Resistance Updates, 14, 203–211, 2011). Experiments have shown that the combination of PIM kinase inhibitors and chemotherapy drugs can significantly improve the efficacy of drug-resistant prostate cancer (Mol. Cancer Ther. 8, 2882–2893, 2009). Therefore, PIM inhibitors can also be used to reverse resistance to chemotherapy.

Experiments have shown that PIM kinase expression is accompanied by activation of T-cells, and PIM-1/3 inhibitors are effective in treating enteritis caused by CD4+ T-cells. The oral clinical trial drug AR452530 can at least reduce rectal inflammation, glandular loss, edema and mucosal hyperplasia by 80% (Cellular Immunology, 272, 200–213, 2012). Therefore, PIM inhibitors can also be used to treat T-cell mediated diseases such as inflammatory bowel disease.

According to the published literature on kinase inhibitors (US Pat. No. 5,952,455 B2):

Both formula A and formula B are PIM inhibitors. When their substituents R _a , R _b and R _c are the same, the only difference is the 5-position substituent on pyridine ring 1, general formula A is a hydrogen atom, and general formula B is a fluorine atom. The literature data shows that in all 29 pairs of compounds, whether the 5-position substituent is H or F has little effect on the PIM activity of these compounds and does not alter the ability of the compound to inhibit PIM-1 enzyme activity.

Summary of the invention

In the study of a new generation of PIM kinase inhibitors, we have unexpectedly found that, unlike known knowledge, in a group of compounds conforming to formula I, when other structures are identical, R ₆ is a cyclic structure. When R _{3 is} changed from H to other substituents, the steric hindrance increases, which hinders the free rotation of the R ₆ ring, thereby increasing their activity. When R _{3 is} replaced by F or CF ₃ , the highest activity is improved by nearly 6 times, an average of 4.19 times, by increasing the lipophilicity of the molecule, thereby further enhancing the ability of the compound to inhibit PIM-1 activity. See Table 2 of Example 57 for specific data.

The technical problem to be solved by the present invention is to study a novel compound of a PIM kinase inhibitor, and to design and prepare a medicament for treating T-cell mediated inflammation such as cancer, multi-directional drug resistance and inflammatory bowel disease.

According to one aspect of the present application, the present invention provides a PIM kinase inhibitor having the formula:

The compound of the formula I according to the invention is a PIM kinase inhibitor of the structure of a pyrimidine compound.

The invention also provides stereoisomers, tautomers and pharmaceutically acceptable salts of the compounds of formula I.

In the above formula I

R ₁ is -H, -NHR ₄ , halogen (F, Cl, Br, I), -OH, -OC _1-3 hydrocarbyl, -SH, -SC _1-3 hydrocarbyl, C _1-3 hydrocarbyl, halo Base, haloethyl, -CN and -NO ₂ ;

R ₂ is -H, -NHR ₄ , halogen (F, Cl, Br, I), -OH, -SH, -OC _1-3 hydrocarbon group with or without a substituent, -SC _1-3 hydrocarbon group, C _1-3 hydrocarbyl and C _3-7 cycloalkyl, halomethyl, haloethyl, -CN and -NO ₂ ;

R ₃ is -NHR ₅ , halogen (F, Cl, Br, I), -OH, -SH, -OC _1-3 hydrocarbon group with or without a substituent, -SC _1-3 hydrocarbon group, C _{1- 3} hydrocarbyl and C _3-7 cycloalkyl, halomethyl, haloethyl, -CN and -NO ₂ ;

R ₄ is -H, -C(=O)-R ₅ , a C _1-8 hydrocarbon group with or without a substituent, a C _3-7 cycloalkyl group, a 4-7 membered heterocyclic group, 5-10 members Aryl and heterocyclic aryl;

R ₅ is a C _1-8 hydrocarbon group, a C _1-8 alkoxy group, a C _3-7 cycloalkyl group with or without a substituent;

R ₆ is a C _3-7 cycloalkyl group, a 4-7 membered heterocyclic group, a 5-10 membered aryl group and a heterocyclic aryl group with or without a substituent; the cyclic hydrocarbon group, an aryl group and a heterocyclic group The substituents on the group may be halogen (F, Cl, Br, I), -CN, -NH ₂ , -NHR ₇ , C _1-4 hydrocarbyl, C _1-4 halohydrocarbyl, C _3-7 cycloalkyl. , -OR ₇ , -NO ₂ , -C(=O)OR ₇ , -C(=O)R ₇ , -C(=O)N(R ₇ ) ₂ , -C(=O)NH ₂ ,- C(=O)NHR ₇ ;

R ₇ is -H or with or without a substituted C _1-4 hydrocarbyl group;

R ₂₂ is a C _1-8 hydrocarbon group with or without a substituent or a group defined by the formula:

Wherein R ₂₃ , R ₂₄ and R ₂₅ are each halogen (F, Cl, Br, I), -OR ₁₅ , -NR ₁₆ R ₁₇ , -C(=O)NR ₁₈ R ₁₉ with or without substitution a C _1-8 hydrocarbon group;

R ₁₅ , R ₁₆ , R ₁₇ , R ₁₈ , R ₁₉ are each -H or a C _1-8 hydrocarbon group with or without a substituent

G ₁ is CH ₂ or N;

G ₂ is NR ₂₈ , CHR ₂₉ or O;

B1 and B2 are 0, 1, 2 or 3;

B3 is 0, 1, 2;

B4 is 0,1;

R ₂₆ and R ₂₇ are each -H or a C _1-8 hydrocarbon group with or without a substituent;

R ₂₈ is -H, a C _1-8 hydrocarbyl group with or without a substituent, a C _3-7 cycloalkyl group, a C _3-7 heterocycloalkyl group, -C(=O)R ₃₀ , -C(=O ) OR ₃₀ or -C(=O)NHR ₃₀ ;

R ₂₉ is -H, -OH, a C _1-8 hydrocarbyl group with or without a substituent, a C _3-7 cycloalkyl group, a C _3-7 heterocycloalkyl group, -NHR ₃₀ , -C(=O)OR ₃₀ or -C(=O)NHR ₃₀ ;

R ₃₀ is -H or a C _1-8 hydrocarbon group with or without a substituent;

The substituent in the present invention may be selected from a hydroxyl group, a nitro group, an amino group, an imino group, a cyano group, a halogen group, a thio group, a sulfonyl group, a thioamido group, a thiol group, or a thiol group, unless otherwise specified. Amidino, oxamidino, methoxamidino, sulfhydryl, sulfonylamino, carboxy, formyl, lower alkyl, halogenated lower alkyl, lower alkylamino, halogenated Lower alkylamino, lower alkoxy, halogenated lower alkoxy, lower alkoxyalkyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, aralkylcarbonyl, heteroaryl.

The "lower", unless otherwise specified, refers to a linear or branched group of _C1-8 , a non-aromatic cyclic group of 3-7 members, or 5-12 yuan, respectively, depending on the defined substituent. An aromatic group, or a 5-12 membered aromatic group having a C _1-8 linear or branched substituent group.

In some embodiments of the invention, the compound of Formula I, and stereoisomers thereof, tautomers and pharmaceutically acceptable salts thereof, R ₁ is -H, -NHR ₄ , halogen (F, Cl, Br, I ), -OH, -SH, -OC _1-3 hydrocarbyl group with or without a substituent, -SC _1-3 hydrocarbyl group, C _1-3 hydrocarbyl group and C _3-7 cycloalkyl group, -CN and -NO ₂ Preferably, R ₁ is -H, -NH ₂ , halogen (F, Cl, Br, I), -OH, -CN and -NO ₂ ; further preferably R ₁ is -H, -NH ₂ , -F.

In another partial embodiment of the invention, the compound of Formula I, and stereoisomers thereof, tautomers and pharmaceutically acceptable salts thereof, R ₂ is -H, -NHR ₄ , halogen (F, Cl, Br, I), -OH, -SH, -OC _1-3 hydrocarbyl group with or without a substituent, -SC _1-3 hydrocarbyl group, C _1-3 hydrocarbyl group and C _3-7 cycloalkyl group, -CN and -NO ₂ ; Preferably, R ₂ is -H, halogen (F, Cl, Br, I), -CN; further preferably R ₂ is -H, halogen (F, Cl, Br, I).

In another partial embodiment of the invention, the compound of Formula I, and stereoisomers thereof, tautomers and pharmaceutically acceptable salts thereof, R ₃ is -NHR ₅ , halogen (F, Cl, Br, I), -OH, -SH, -OC _1-3 hydrocarbyl group with or without a substituent, -SC _1-3 hydrocarbyl group, C _1-3 hydrocarbyl group and C _3-7 cycloalkyl group, halomethyl group, halogenated B a group, -CN and -NO ₂ ; preferably R ₃ is halogen (F, Cl, Br, I), a C _1-3 hydrocarbon group with or without a substituent, and a -OC _1-3 hydrocarbon group, a halogenated methyl group , -CN and -NO ₂ ; further preferably R ₃ is halogen (F, Cl, Br, I), -CF ₃ , and -CN; still more preferably R ₃ is halogen (F, Cl, Br, I) and - CF ₃ .

In another embodiment of the invention the part, the I compounds of the general formula and stereoisomers thereof, tautomers thereof and pharmaceutically acceptable salts thereof, R ₆ is with or without a C _3-7 cycloalkyl group substituted hydrocarbyl a 4-7 membered heterocyclic group, a 5-10 membered aryl group and a heterocyclic aryl group; the substituents on the cycloalkyl, aryl and heterocyclic aryl groups may be halogen (F, Cl, Br, I), respectively , -CN, -NH ₂ , -NHR ₇ , C _1-4 hydrocarbyl, C _1-4 halohydrocarbyl, C _3-7 cycloalkyl, -OR ₇ , -NO ₂ , -C(=O)OR ₇ , -C(=O)R ₇ , -C(=O)N(R ₇ ) ₂ , -C(=O)NH ₂ , -C(=O)NHR ₇ ; preferably R ₆ is phenyl, piperazinyl And a pyridyl group, the phenyl group, piperazinyl group and pyridyl group may be selected from 1-3 selected from -F, -Cl, -Br, -I, -OH, -NH ₂ , C _1-3 hydrocarbon group, C ₁ Substituted with a _-3 alkoxy group, a halogenated C _1-3 hydrocarbyl group; further preferably R ₆ is phenyl, pyridyl, 2-F-6-OCH ₃ -phenyl, 2,6-difluorophenyl .

In another partial embodiment of the present invention, in the compound of the formula I, and the stereoisomers, tautomers and pharmaceutically acceptable salts thereof, it is preferred that R ₂₂ is a cyclobutane group with or without a substituent. Cyclopentyl, cyclohexane, cycloheptyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, butylene oxide, tetrahydrofuranyl, tetrahydropyran Further preferably, R ₂₂ is a cyclohexane group, a pyrrolidinyl group, a piperidinyl group or an azacycloheptyl group; and the substituent group may have 1 to 4 substituents, which may be selected separately From halogen (F, Cl, Br, I), -NH ₂ , -OH, methylamino, ethylamino, propylamino, dimethylamino, diethylamino, methyl, ethyl, propyl, methyl Oxyl, ethoxy, propoxy, monohalomethyl, polyhalomethyl, monohaloethyl, polyhaloethyl; preferred substituents are halogen (F, Cl, Br, I), -NH ₂ , -OH, methylamino, methyl and methoxy; further preferred substituents are -NH ₂ , -OH and methyl.

In another partial embodiment of the invention, the compound of Formula I, and stereoisomers, tautomers thereof, and pharmaceutically acceptable salts thereof, wherein _R22 is cyclobutanemethyl with or without a substituent , cyclopentylmethyl, cyclohexanemethyl, cycloheptylmethyl, azetidinylmethyl, pyrrolidinylmethyl, piperidinylmethyl, azepanylmethyl , butylene alkylmethyl, tetrahydrofuranylmethyl, tetrahydropyranylmethyl; further preferably R ₂₂ is cyclohexanemethyl, pyrrolidinylmethyl, piperidinylmethyl, azepan An alkylmethyl group; may have from 1 to 4 substituents on the substituent group, and may be independently selected from the group consisting of halogen (F, Cl, Br, I), -NH ₂ , -OH, methylamino group ,ethylamino, propylamino, dimethylamino, diethylamino, methyl, ethyl, propyl, methoxy, ethoxy, propoxy, monohalomethyl, polyhalomethyl , monohaloethyl, polyhaloethyl; preferred substituents are halogen (F, Cl, Br, I), -NH ₂ , -OH, methylamino, methyl and methoxy; further preferred substituents It is -NH ₂ , -OH and methyl.

In another partial embodiment of the invention, the compound of Formula I, and the stereoisomers, tautomers and pharmaceutically acceptable salts thereof, wherein _R22 is a _C2-5 hydrocarbon group with or without a substituent, These hydrocarbon groups may have up to 4 substituents, which may be halogen (F, Cl, Br, I), NH ₂ , methylamino, ethylamine, propylamino, dimethylamino, diethylamino, respectively. Hydroxy, methyl, ethyl, propyl, -CH ₂ OH, -CH ₂ (OH)CH ₃ , -CH ₂ CH ₂ OH, monohalomethyl, polyhalomethyl, monohaloethyl, Polyhaloethyl, C _1-4 hydrocarbyl-O-, C _1-4 hydrocarbyl-S-.

In some embodiments of the invention, the compounds of formula I, and their stereoisomers, tautomers and pharmaceutically acceptable salts thereof, are shown in Table 2.

Definition of terms used in this article:

The term "alkyl" as used herein, refers to an alkyl group that does not contain a hetero atom. Thus, the term includes straight-chain alkyl groups such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, decyl, decyl, undecyl, dodecyl, and the like. . The term also includes branched chain isomers of straight chain alkyl groups, including but not limited to, the following _{groups: -CH (CH 3) 2,} CH (CH 3) (CH 2 CH 3), CH (CH 2 CH ₃ ) ₂ , -C(CH ₃ ) ₃ , -C(CH ₂ CH ₃ ) ₃ , -CH ₂ CH(CH ₃ ) ₂ , -CH ₂ CH(CH ₃ )(CH ₂ CH ₃ ), -CH ₂ CH(CH ₂ CH ₃ ) ₂ , -CH ₂ C(CH ₃ ) ₃ , -CH ₂ C(CH ₂ CH ₃ ) ₃ , -CH(CH ₃ )CH(CH ₃ )(CH ₂ CH ₃ ), - CH ₂ CH ₂ CH(CH ₃ ) ₂ , -CH ₂ CH ₂ CH(CH ₃ )(CH ₂ CH ₃ ), -CH ₂ CH ₂ CH(CH ₂ CH ₃ ) ₂ , -CH ₂ CH ₂ C(CH ₃ ) ₃ , -CH ₂ CH ₂ C(CH ₂ CH ₃ ) ₃ , -CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ , -CH(CH ₃ )CH(CH ₃ )CH(CH ₃ ) ₂ And CH(CH ₂ CH ₃ )CH(CH ₃ )CH(CH ₃ )(CH ₂ CH ₃ ). Thus, the term alkyl includes primary alkyl, secondary alkyl and tertiary alkyl. Preferred alkyl groups include straight chain and branched alkyl groups having from 1 to 12 carbon atoms. A preferred "alkyl group" relates to the definition of straight chain C _l-4 alkyl such as methyl, ethyl, n-propyl and n-butyl. Preferred alkyl groups include C _3-5 define further branched chain alkyl group, including _{-CH (CH 3) 2, -CH} 2 CH (CH 3) 2, -CH (CH 3) CH 2 CH 3, -C (CH ₃ ) ₃ , -CH(CH ₃ )CH ₂ CH ₂ CH ₃ , -CH(CH ₃ )CH(CH ₃ ) ₂ , -CH ₂ CH(CH ₃ )CH ₂ CH ₃ , -CH ₂ CH ₂ CH ( CH ₃ ) ₂ and -CH(CH ₂ CH ₃ ) ₂ and the like.

The term "alkenyl" as used herein, refers to an alkyl group as defined above wherein at least one point of unsaturation, i.e., wherein two adjacent carbon atoms are joined by a double bond.

The term "alkynyl" as used herein relates to an alkyl group wherein two adjacent carbon atoms are joined by a triple bond.

The term "alkoxy" as used herein, refers to -OR, wherein R is alkyl.

The term "hydrocarbyl" as used herein is a generic term for alkyl, alkenyl and alkynyl groups.

The term "halogen" or "halo" as used herein, refers to a fluorine, chlorine, bromine and iodine (F, Cl, Br, I) group. "Haloalkyl" means an alkyl group substituted by one or more halogen atoms. Thus, the term "haloalkyl" includes monohaloalkyl, dihaloalkyl, trihaloalkyl and the like. Typical monohaloalkyl groups include -CH ₂ F, -CH ₂ Cl, -CH ₂ CH ₂ F, -CH ₂ CH ₂ Cl, -CH(F)CH ₃ , -CH(Cl)CH _{3 ;} typical dihaloalkanes groups include _{-CHC1 2, -CHF 2, -CC1 2} CH 3, -CH (Cl) CH 2 Cl, -CH 2 CHC1 2, -CH 2 CHF 2; trihaloalkyl typically comprises -CC1 _3, -CF ₃ , -CC1 ₂ CH ₂ Cl, -CF ₂ CH ₂ F, -CH(Cl)CHC1 ₂ , -CH(F)CHF ₂ ; typical perhaloalkyl groups include -CC1 ₃ , -CF ₃ , -CC1 ₂ CC1 ₃ , -CF ₂ CF ₃ .

As used herein, "amino" refers to the group NH _2.

The term "alkylamino" as used herein refers to a group -NRR' wherein R and R' are each independently selected from hydrogen or lower alkyl, wherein R and R' are not H at the same time.

The term "arylamino" as used herein refers to a group -NRR' wherein R is aryl and R' is hydrogen, lower alkyl or aryl.

The term "aralkylamino" as used herein refers to a group -NRR' wherein R is lower aralkyl and R' is hydrogen, lower alkyl, aryl or lower aralkyl.

The term cyano, as used herein, refers to the group -CN.

The term nitro as used herein refers to the group -NO ₂ .

The term "alkoxyalkyl" as used herein refers to a group -alk1-0-alk2 wherein alk1 is alkyl or alkenyl and alk2 is alkyl or alkenyl. The term "lower alkoxyalkyl" refers to an alkoxyalkyl group wherein alk1 is lower alkyl or lower alkenyl and alk2 is lower alkyl or lower alkenyl. The term "aryloxyalkyl" refers to the group monoalkyl-0-aryl. The term "aralkyloxyalkyl" refers to a group of alkylene-0-aralkyl wherein the aralkyl group is a lower aralkyl group.

The term "aminocarbonyl" as used herein refers to the group -C(=O)-NH ₂ ; "substituted aminocarbonyl" refers herein to a group wherein R is lower alkyl and R' is hydrogen or lower alkyl. Group C(O)-NRR'. In some embodiments, R and R' may together with the N atom to which they are attached form a "heterocycloalkylcarbonyl." The term "arylaminocarbonyl" refers herein to a group -C(O)-NRR' wherein R is aryl and R' is hydrogen, lower alkyl or aryl. "Aralkylaminocarbonyl" refers herein to a group -C(O)-NRR' wherein R is lower aralkyl and R' is hydrogen, lower alkyl, aryl or lower aralkyl.

As used herein, "aminosulfonyl" refers to the group -S(O) ₂ NH ₂ "substituted aminosulfonyl" as used herein to refer to a group wherein R is lower alkyl and R' is hydrogen or lower alkyl. -S(O) ₂ NRR'. The term "aralkylaminosulfonylaryl" refers herein to the group aryl-S(O) _2- NH aralkyl wherein the aralkyl group is a lower aralkyl group.

As used herein, "carbonyl" refers to the divalent group -C(O)-. "Carboxyl" means -C(=O)-OH. "Alkoxycarbonyl" means an ester-C(=O)-OR wherein R is alkyl. "Lower alkoxycarbonyl" means an ester-C(=O)-OR wherein R is lower alkyl. "Cycloalkoxycarbonyl" means C(=O)-OR wherein R is cycloalkyl.

As used herein, "cycloalkyl" refers to a mono or polycyclic carbocyclic alkyl substituent. Carbocycloalkyl is a cycloalkyl group in which all ring atoms are carbon. Typical cycloalkyl substituents have from 3 to 8 backbone (ie, ring) atoms wherein each backbone atom is a carbon or heteroatom. The term "heterocycloalkyl" as used herein refers to a cycloalkyl substituent having from 1 to 5, and more typically from 1 to 4, heteroatoms in the ring structure. Suitable heteroatoms for use in the compounds of the invention are nitrogen, oxygen and sulfur. Representative heterocycloalkyl moieties include, for example, morpholino, piperazinyl, piperidinyl and the like. Carbocycloalkyl is a cycloalkyl group in which all ring atoms are carbon. The term "polycyclic" as used herein, when used in connection with a cycloalkyl substituent, refers to both fused and non-fused alkyl cyclic structures. The terms "partially unsaturated cycloalkyl", "partially saturated cycloalkyl" and "cycloalkenyl" all mean that there is at least one point of unsaturation, ie wherein two adjacent ring atoms pass through a double bond or three A cycloalkyl group bonded to a bond. Illustrative examples include cyclohexynyl, cyclopentynyl, cyclopropenyl, cyclobutane, and the like.

The term "substituted heterocyclic ring", "heterocyclic group" or "heterocyclic ring" as used herein, refers to any 3 or 4 membered ring containing one hetero atom selected from nitrogen, oxygen and sulfur or containing one to three selected from one to three. a 5- or 6-membered ring of a hetero atom of nitrogen, oxygen or sulfur; wherein the 5-membered ring has 0-2 double bonds and the 6-membered ring has 0-3 double bonds; wherein the nitrogen and sulfur atoms are Optionally oxidized; and includes any bicyclic group wherein any of the above heterocyclic rings are fused to a phenyl ring or other 5- or 6-membered heterocyclic ring as defined above independently. The term "heterocycloalkyl" as used herein, refers to a 5- or 6-membered ring containing one to three heteroatoms selected from nitrogen, oxygen or sulfur, wherein the ring has no double bonds. For example, the term heterocyclic- _C5 alkyl refers to a 6-membered ring containing 5 carbon atoms and one hetero atom such as N. Thus, the term "heterocycle" includes rings wherein the nitrogen is a heteroatom and a partially saturated and fully saturated ring. Preferred heterocycles include, for example, dinitrogen

(diazapiny1), pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, pyridyl, piperidinyl, pyridazinyl, piperazinyl, pyrazinyl, N- Methyl piperazinyl, azetidinyl, N-methylazetidinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl , morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, fluorenyl, quinolyl, isoquinolinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, furanyl a thienyl group, a triazolyl group and a benzothienyl group; the heterocyclic moiety may be unsubstituted or monosubstituted or disubstituted or trisubstituted by various substituents independently selected from the group consisting of a hydroxyl group, a halogen, and an oxo group (C). =O), alkylimino (RN=, wherein R is lower alkyl or lower alkoxy), amino, alkylamino, dialkylamino, acylaminoalkyl, alkoxy, thioalkoxy , polyalkoxy, lower alkyl, cycloalkyl or haloalkyl.

It will be readily understood by those skilled in the art of organic and medicinal chemistry in conjunction with the disclosure herein that heterocyclic groups can be attached at a variety of positions. Typical heterocycles include, for example, imidazolyl, pyridyl, piperazinyl, piperidinyl, azetidinyl, thiazolyl, furyl, triazolyl, benzimidazolyl, benzothiazolyl, benzimidin Azolyl, quinolyl, isoquinolyl, thiazolinyl, quinazolinyl, pyridazinyl, fluorenyl, naphthyridinyl, oxazolyl and quinazolyl.

As used herein, "aryl" refers to optionally substituted monocyclic and polycyclic aromatic groups having from 3 to 14 backbone carbon or heteroatoms, and includes carbocyclic aryl and heterocyclic aryl. A carbocyclic aryl group is an aryl group in which all of the ring atoms in the aromatic ring are carbon. The term "heteroaryl" refers herein to an aryl group having from 1 to 4 heteroatoms in the aromatic ring as a ring atom and the remaining ring atoms being carbon atoms. When used in combination with an aryl substituent, the term "polycyclic aryl" refers herein to a fused and non-fused cyclic structure in which at least one ring structure is aromatic, such as a benzodioxolane ring. (It has a heterocyclic structure fused to a phenyl group, that is, a naphthyl group or the like). Examples of the aryl moiety used as a substituent in the compound of the present invention include phenyl, pyridyl, pyrrolyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl, tetrazolyl, pyrazinyl, triazole A group, a thienyl group, a furyl group, a quinolyl group, a fluorenyl group, a naphthyl group, a benzothiazolyl group, a benzopyridyl group, a benzimidazolyl group, and the like.

As used herein, "optionally substituted" or "substituted" means that one or more hydrogen atoms are replaced by a monovalent or divalent group; suitable substituent groups include, for example, hydroxy, nitro, amino, imino, cyano , halogen, thio group, sulfonyl group, thioamido, sulfhydryl, fluorenylene, oxamidino, methoxamidino, sulfhydryl, sulfonylamino, Carboxyl, formyl, lower alkyl, halogenated lower alkyl, lower alkylamino, halogenated lower alkylamino, lower alkoxy, halogenated lower alkoxy, lower alkoxyalkyl, alkylcarbonyl, Aminocarbonyl, arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl, heteroarylalkylcarbonyl, alkylthio, aminoalkyl, cyanoalkyl, aryl, etc.; substituents may be substituted; substituted to substituted The group on the group may be carboxyl, halogen, nitro, amino, cyano, hydroxy, lower alkyl, lower alkoxy, aminocarbonyl, SR, thioamido, SO ₃ H, SO ₂ R or naphthe a group, wherein R is usually hydrogen, an amide or a lower alkyl; when the substituted substituent includes a linear group , The substituent may be located at the end of the chain or within the chain (e.g., 2-hydroxypropyl, 2-aminobutyl, and the like) (e.g., 2-hydroxyethyl, 3-aminopropyl, etc.). Substituted substituents can be linear, branched or cyclic arrangements of covalently bonded carbon or heteroatoms. The above definition does not include impermissible substitution patterns (for example, a methyl group substituted by five fluorine groups or a halogen atom substituted by another halogen atom); such a mode of substitution which is not allowed is well known to those skilled in the art. .

It will also be apparent to those skilled in the art that the compounds of the invention or their stereoisomers, as well as pharmaceutically acceptable salts, esters, metabolites and prodrugs thereof, can be tautomerized and thus may be The various tautomeric forms in which the protons of one atom of a molecule are shifted to another atom and the chemical bonds between the atoms of the molecule are thus rearranged. See, for example, March, Advanced Organic Chemistry: Reactions, Mechanisms and Structures, Fourth Edition, John Wiley & Sons, pp. 69-74 (1992). The term "tautomer" as used herein refers to a compound produced by proton shift, and it should be understood that all tautomeric forms are included in the present invention as long as it is possible.

The compounds of the invention or their tautomers, and pharmaceutically acceptable salts, esters, metabolites and prodrugs thereof, may comprise asymmetrically substituted carbon atoms. Such asymmetrically substituted carbon atoms can give rise to the compounds of the invention in enantiomers, diastereomers and other stereoisomeric forms which can be defined in terms of absolute stereochemistry, such as (R)- Or (S)-form. Thus, all such possible isomers of the compounds of the invention, their individual stereoisomers in optically pure form, mixtures thereof, racemic mixtures (or "racemates"), diastereomers Mixtures as well as individual diastereomers are included in the present invention. The terms "S" and "R" configurations as used herein are as defined in IUPAC 1974 RECOMMENDATIONS FOR SECTION E, FUNDAMENTAL STEREOCHEMISTRY, Pure Appl. Chem. 45: 13-30 (1976). The term and ring position for a cyclic compound. The - side of the reference face is the side on which the preferred substituent is located at the lower numbered position. Those substituents located on the reference facing side are described by themselves. It should be noted that this usage is different from the usage for the annular stereoparents, in the latter case, "" means "below the plane" and represents the absolute configuration. The terms and configurations used herein are as defined in paragraph 203 of CHEMICALABSTRACTS INDEX GUIDE-APPENDIX IV (1987).

The term "pharmaceutically acceptable salt" as used herein refers to a non-toxic acid or alkaline earth metal salt of a compound of formula I. These salts can be prepared in situ during the final isolation and purification of the compound of formula I, or can be prepared by separately reacting a base or acid functional group with a suitable organic or inorganic acid or base. Typical salts include, but are not limited to, the following salts: acetate, adipate, alginate, citric acid Salt, aspartate, benzoate, besylate, hydrogen sulfate, butyrate, camphorate, sulfonate, digluconate, cyclopentane propionate, dodecyl Sulfate, ethanesulfonate, glucoheptonate, glycerol phosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyl Ethyl sulfonate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, pamoate, alanate, persulfate, 3-benzene Propionate, picrate, pivalate, propionate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate. Reagents such as lower alkyl halides such as methyl, ethyl, propyl and butyl chloride, bromide and iodide; dialkyl sulfates such as dimethyl sulfate, diethyl ester, dibutyl ester may also be used. And diamyl esters, long chain halides such as decyl, lauryl, myristyl and stearyl chloride, bromide and iodide, aralkyl halides such as benzyl bromide and phenethyl bromide will contain The basic nitrogen group is quaternized. Water or oil soluble or water or oil dispersible products are thus obtained.

Examples of the acid which can be used to form a pharmaceutically acceptable acid addition salt include inorganic acids such as hydrochloric acid, sulfuric acid and phosphoric acid, and organic acids such as oxalic acid, maleic acid, methanesulfonic acid, succinic acid and citric acid. The base addition salt can be prepared in situ during the final isolation and purification of the compound of formula (I), or can be obtained by reacting a carboxylic acid moiety with a suitable base such as a hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation. It can be prepared by reacting ammonia or an organic primary, secondary or tertiary amine. Pharmaceutically acceptable salts include, but are not limited to, alkali and alkaline earth metal based cations such as sodium, lithium, potassium, calcium, magnesium, aluminum salts, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations including, but not limited to, ammonium. a salt of tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine or ethylamine. Other typical organic limbs used to form base addition salts include diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like.

The term "pharmaceutically acceptable ester" as used herein refers to esters which hydrolyze in vivo, including those which are readily decomposed in the human body to release the parent compound or a salt thereof. Suitable esters include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids, particularly alkanoic acids, alkenoic acids, naphthenic acids and alkanoic acids, wherein each alkyl or alkenyl moiety preferably has No more than 6 carbon atoms. Examples of specific esters include formates, acetates, propionates, butyrates, acrylates, and ethylsuccinates.

The term "pharmaceutically acceptable prodrug" as used herein means that it is suitable for contact with tissues of humans and lower animals in a reasonable medical judgment without excessive toxicity, irritation, allergic reaction, etc., with reasonable benefits/risks. Prodrugs of the compounds of the invention, as well as the zwitterionic forms of the compounds of the invention, which are effective, and which are effective for their desired application. The term "prodrug" refers to a compound that is rapidly converted in vivo, for example by hydrolysis in blood, to produce the parent compound of the above formula. In T. Higuchi and v. Stella, as a Pro-drugs as Novel Delivery Systems, Vol.14, ACSSymposium Series and Edward B. Roche, Bioreversible Vectors in Drug Design ( A detailed discussion is provided in Bioreversible Carriers in Drug Design, both of which are incorporated herein by reference.

Any structural formula given herein also represents an unlabeled form as well as an isotopically labeled form of the compound. Isotopically labeled compounds have the structure described by the structural formula shown herein, except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes which may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as ² H, ³ H, ^ll C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ F , ³¹ P, ³² P, ³⁵ S, ³⁶ Cl, ¹²⁵ I. The invention includes various isotopically-labeled compounds of the present invention, such as those in which radioactive isotopes such as ³ H and ¹⁴ C are present. Such isotopically labeled compounds can be used in metabolic studies (using ¹⁴ C), reaction kinetic studies (using, for example, ² H or ³ H), detection or imaging techniques such as positron emission tomography (PET) or single photon emission computers. Tomography (SPECT), including drug or substrate tissue distribution tests, or can be used in the radiotherapy of patients. In particular, ¹⁸ F or labeled compounds are particularly useful for PET or SPECT studies. Isotopically labeled compounds of the invention, as well as prodrugs thereof, can generally be prepared by the practice procedures or procedures disclosed in the Examples and Preparations below, by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent. In addition, replacement with heavier isotopes, particularly deuterium (ie, ² H or D), may provide certain therapeutic benefits resulting from increased metabolic stability, such as increased in vivo half-life or reduced dosage or improved therapeutic index. . It should be clear that ruthenium in this context is considered to be a substituent of the compound of formula (1). The concentration of such heavier isotopes, especially strontium, can be defined by isotopic enrichment factors. The term "isotopic enrichment factor" as used herein refers to the ratio between the isotope abundance and the natural abundance of a given isotope. If a substituent in the compound of the present invention is designated as hydrazine, the compound has at least 3,500 (52.5% of ruthenium mixed in each designated atom) and at least 4,000 (60% of ruthenium mixed) for each of the specified ruthenium atoms. At least 4,500 (67.5% of cesium mixed), at least 5,000 (75% of cesium mixed), at least 5,500 (82.5% of cesium mixed), at least 6,000 (90% of cockroach mixed), at least 6333.3 (95 % 氘 mixed), at least 6466.7 (97% mash mixed), at least 6600 (99% mash mixed) or at least 6633.3 (99.5% mash mixed) isotope enrichment factor.

Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or can be replaced by suitable isotopically labeled reagents by methods analogous to those described in the accompanying examples and preparations. The non-isotopically labeled reagents used were prepared.

It will be apparent to those skilled in the art that the compounds of the present invention, or their tautomers, prodrugs and stereoisomers, and pharmaceutically acceptable salts, esters and prodrugs thereof, can be passed It is metabolized in the human or animal body or cells and processed in the body to produce metabolites. The term "metabolite" as used herein refers to any structural derivative produced in an individual after administration of the parent compound. These derivatives can be produced from the parent compound by various biochemical transformations such as oxidation, reduction, hydrolysis or binding in the body, and include, for example, oxides and demethylated derivatives. Metabolites of the compounds of the invention can be identified using conventional techniques known in the art. See, for example, Bertolini, G. et al.,]. Med. Chem. 40: 2011-2016 (1997); Shan, D. et al., J. Pharm. Sci. 86(7): 765-767; Bagshawe K., Drug Dev. Res. 34: 220-230 (1995) Bodor, N., Advances in Drug Res. 13:224-331 (1984); Bundgaard, H., Design of Prodrugs (Elsevier Press 1985); and Larsen, 1.K., Design and Application of Prodrugs, Drug Design And Development (Krogsgaard-Larsen et al., Harwood Academic Publishers, 1991). It should be clear that the chemical compounds of the compounds of the formula I or their tautomers, prodrugs and stereoisomers, as well as the pharmaceutically acceptable salts, esters and prodrugs of any of them Both are included in the present invention.

The term "cancer" as used herein refers to a cancerous disease which can be beneficially treated by inhibiting PIM kinase, including, for example, including, for example, solid tumors such as lung cancer, pancreatic cancer, thyroid cancer, ovarian cancer, bladder cancer, breast cancer, prostate cancer or colon. Cancer, melanoma, bone marrow disorders such as myeloid leukemia, multiple myeloma and erythroleukemia, adenomas, for example, villous colon adenomas and sarcomas such as osteosarcoma; liquid tumors such as chronic lymphocytic leukemia, acute lymphocytes Leukemia, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, and multiple myeloma.

The PIM kinase inhibitors provided by the present invention include the following compounds:

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

N-(4-(azetidin-3-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

N-(4-((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Amide

N-(4-((3-Aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

N-(4-(3-Aminopropoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(3-(methylamino)propoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-N-(4-(3-(dimethylamino)propoxy)pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide

N-(4-(4-Aminobutoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(4-(methylamino)butoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(3-hydroxypropoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-difluorophenyl)-5-fluoro-N-(4-(3-hydroxybutoxy)pyrimidin-5-yl)pyridine amide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(4-hydroxybutoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-((4-hydroxy-4-methylpentyloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-N-(4-(3,4-dihydroxybutoxy)pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-[4-(tetrahydropyran-4-methoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidin-5-yl)pyridine amide

3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

3-Amino-N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine A Amide

3-amino-N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro Pyridylamide

3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

N-(4-((1-Amino-3-chloropropyl-2-yl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2 -pyridinecarboxamide

N-(4-(3-Amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine A Amide

3-amino-N-(4-(azetidin-3-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Amide

3-amino-N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Formamide

3-amino-N-(4-((1-amino-3-chloropropyl-2-yl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)- 5-fluoro-2-pyridinecarboxamide

3-amino-N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro -2-pyridinecarboxamide

3-amino-5-fluoro-6-phenyl-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

3-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-(2,4'-bipyridyl)-6-carboxamide

N-(4-(azetidin-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-3-fluoro-2-pyridinecarboxamide

N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)-2-pyridinecarboxamide

N-(4-(azetidin-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidine-5-trifluoromethyl-2-pyridinecarboxamide

N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxamide

6-(2,6-Difluorophenyl)-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridinecarboxamide

N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2 -pyridinecarboxamide

6-(2,6-Difluorophenyl)-N-[4-(tetrahydropyran-4-yl)methoxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridine Amide

3-amino-N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidine-5-trifluoromethyl-2-pyridine Formamide

3-Amino-N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl- 2-pyridinecarboxamide

3-amino-6-(2,6-difluorophenyl)-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridine Formamide

3-Amino-N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-III Fluoromethyl-2-pyridinecarboxamide

3-amino-6-phenyl-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridinecarboxamide

N-(4-(3-Amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2 -pyridinecarboxamide

3-amino-N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-III Fluoromethyl-2-pyridinecarboxamide

N-(4-(3-Amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Amide

3-amino-N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro -2-pyridinecarboxamide

N-(4-(3-Amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2 -pyridinecarboxamide

3-amino-N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-three Fluoromethyl-2-pyridinecarboxamide

Another object of the present invention is to provide a process for the preparation of the above PIM kinase inhibitor.

The compounds of the invention are prepared starting from commercially available starting materials and reagents. The method of the present invention is represented by the following formula:

Alternatively, the alcohol B (1.1 equivalents) with or without a protecting group is first reacted with a base such as NaH (1.1 equivalents) in a solvent such as tetrahydrofuran at room temperature (25 ° C) for 1 hour, then Further, an aminopyrimidine ether C is obtained by reacting with 5-amino-4-chloropyrimidine A (1 equivalent) under heating at, for example, 100 ° C for 1 to 10 hours. Aromatic carboxylic acid D (1 equivalent) with or without a protecting group in the presence of a coupling reagent such as HATU (1.1 equivalents), a base such as DIEA (3 equivalents) in a solvent such as DMF and amine C ( 1 equivalent) was reacted to give an ether compound E. If there is no protecting group in E, then E is an ether compound of the formula I in which the E substituent is a hydrocarbyloxy group. If with a protecting group, such as BOC (tert-butoxyformate) or trimethylsilane, E with 10 to 100 equivalents of trifluoroacetic acid and an equal volume of dichloromethane, or 2 equivalents of hydrochloric acid After the methanol solution was mixed at room temperature (25 ° C), the mixture was stirred for 1 to 16 hours, and then the solvent was distilled off under reduced pressure at room temperature (25 ° C) to give an ether compound of the formula I in which the E substituent was a hydrocarbyloxy group.

A further object of the present invention is to provide the use of the above PIM kinase inhibitors in pharmaceuticals.

All of the compounds in the examples were significantly inhibited by the PIM kinase biochemical activity assay, and the IC50 was in the range of 0.1-50 nM. Therefore, the PIM kinase inhibitor of the present invention can be used for the preparation of a medicament.

According to another aspect of the present application, the present invention provides the use of the above PIM kinase inhibitor for the preparation of a medicament for treating or preventing an autoimmune disease.

According to another aspect of the present application, the present invention provides the use of the above PIM kinase inhibitor for the preparation of a medicament for treating or preventing an allergic disease.

According to another aspect of the present application, the present invention provides the use of the above PIM kinase inhibitor for the preparation of a medicament for treating or preventing atherosclerosis.

According to another aspect of the present application, the present invention provides the use of the above PIM kinase inhibitor for the preparation of a medicament for treating or preventing an anti-organ transplant rejection.

According to another aspect of the present application, the invention provides the use of the above PIM kinase inhibitor for the preparation of a medicament for treating or preventing cancer.

According to another aspect of the present application, the present invention provides the use of the above PIM kinase inhibitor for the preparation of a medicament for treating or preventing multi-directional drug resistance.

According to another aspect of the present application, the present invention provides the use of the above PIM kinase inhibitor for the preparation of a medicament for treating or preventing T cell mediated inflammation.

According to another aspect of the present application, the medicament of the present invention is a pharmaceutical composition comprising a PIM kinase inhibitor as an active ingredient and a pharmaceutically acceptable carrier.

According to another aspect of the present application, the present invention provides a novel use of a PIM kinase inhibitor, which has great clinical application value.

According to another aspect of the present application, the compounds and pharmaceutical compositions of the present invention are useful for treating or preventing cancer, reversing anticancer and other drug resistance, inflammatory bowel disease, autoimmune diseases, allergic diseases, arterial porridge Sclerosis, anti-organ transplant rejection, multi-directional drug resistance, T cell-mediated inflammation; "cancer" refers to cancerous diseases that can be beneficially treated by inhibiting PIM kinase, including, for example, solid tumors such as lung cancer, pancreas Cancer, thyroid cancer, ovarian cancer, bladder cancer, breast cancer, prostate cancer or colon cancer, melanoma, bone marrow disorders such as myeloid leukemia, multiple myeloma and erythroleukemia, gland Tumors such as, villous colon adenomas and sarcomas such as osteosarcoma; liquid tumors such as chronic lymphocytic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, and multiple myeloma .

According to another aspect of the present application, the present invention provides the compound and the pharmaceutical composition described above for the preparation of a medicament for treating or preventing cancer, reversing anti-cancer and other drug resistance, inflammatory bowel disease, autoimmune disease, allergic reaction disease, Atherosclerosis, anti-organ transplant rejection, multi-directional resistance, T cell-mediated inflammation; "cancer" refers to cancerous diseases that can be beneficially treated by inhibiting PIM kinase, including, for example, solid tumors such as lung cancer , pancreatic cancer, thyroid cancer, ovarian cancer, bladder cancer, breast cancer, prostate cancer or colon cancer, melanoma, bone marrow disorders such as myeloid leukemia, multiple myeloma and erythroleukemia, adenomas, for example, villous colon glands Tumors and sarcomas such as osteosarcoma; liquid tumors such as chronic lymphocytic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, non-Hodgkin's lymphoma, and multiple myeloma drugs.

detailed description

The technical solutions of the present application are clearly and completely described in the following with reference to the embodiments. It is obvious that the described embodiments are a part of the embodiments of the present application, and not all of the embodiments. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present application without departing from the inventive scope are the scope of the present application.

Synthesis of intermediate 2-picolinic acid (D)

1. Synthesis of 6-(2,6-difluorophenyl)-5-fluoro-2-picolinic acid (D1)

Synthesis of (i) 2-(2,6-difluorophenyl)-3-fluoro-6-methylpyridine (c)

2-Bromo-3-fluoro-6-methylpyridine (a) (279 mg, 1.77 mmol), 2,6-difluorophenylboronic acid (b) (167 mg, 0.88 mmol) and potassium fluoride (169 mg, 2.91) Ment) dissolved in tetrahydrofuran/water mixed solvent (10:1, 8 ml), degassed for 5 min, added Pd ₂ (dba) ₃ (25 mg, 0.044 mmol) and tri-tert-butylphosphine (18 mg, 0.088 mmol) under nitrogen. The mixture was heated to 60 ° C, and reacted for 1 hour. The mixture was cooled to room temperature. ethyl acetate (10 ml) was added, washed with water (10 ml) and brine (10 ml), dried over anhydrous sodium sulfate The product was dissolved in ethanol (10 ml), and then added with NaBH4 (17 mg, 0.44 mmol), and the mixture was stirred at room temperature for 1 hour, 10 ml of water, and then extracted with diethyl ether (3×10 ml), and the combined organic layers were washed with saturated brine (10 ml) Drying with sodium sulfate and removing the solvent gave a crude product. The crude product was purified with EtOAc EtOAc (EtOAc:EtOAc:

(ii) Synthesis of 6-(2,6-difluorophenyl)-5-fluoro-2-picolinic acid (D1)

The compound c (300 mg, 1.34 mmol) and potassium permanganate (423 mg, 2.68 mmol) were dissolved in 45 ml of water and refluxed for 24 hours. After cooling, the mixture was filtered over Celite, and then filtered, The solid was extracted with ethyl acetate (3×50 ml). The mixture was concentrated to 10 ml, and then extracted with 1N sodium hydroxide solution (3×5 ml), and the mixture was neutralized with concentrated hydrochloric acid to pH=1 to give a solid. The solids were combined twice to give the product D1 (153 mg, 0.603 mmol)

2. Synthesis of 3-amino-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxylic acid (D2)

Synthesis of (i) 3-Amino-5-fluoro-2-picolinic acid methyl ester (f)

3-Amino 2-bromo-5-fluoropyridine (500 mg, 2.62 mmol), triethylamine (0.59 ml, 4.16 mmol), Pd(BINAP)Cl ₂ (32 mg, 0.039 mmol), Water methanol (20 ml), degassed, charged with carbon monoxide gas (60 psi), heated to 100 ° C, reacted for 12 hours, cooled to room temperature, then added Pd (BINAP) Cl ₂ (10 mg, 0.012 mmol), continued at 60 psi carbon monoxide, After reacting at 100 ° C for 12 hours, it was cooled to room temperature and filtered. Ethyl acetate (30 ml) was added toEtOAc.

(ii) Synthesis of methyl 3-amino-6-bromo-5-fluoro-2-picolinate (g)

Methyl 3-amino-5-fluoro-2-picolinate (100 mg, 0.588 mmol) was dissolved in acetonitrile (2 mL). NBS (105 mg, 0.647 mmol) reaction. The reaction mixture was extracted with EtOAc (EtOAc m. The crude product obtained after concentration of the filtrate was purified by silica gel chromatography (20-60% ethyl acetate / petroleum ether) to give product (g) (59mg, 0.235mmol)

(iii) Synthesis of methyl 3-amino-6-(2,6-difluorophenyl)-5-fluoro-2-picolinic acid (h)

Referring to the synthesis method of the compound c, the product (h) is obtained by using methyl 3-amino-6-bromo-5-fluoro-2-picolinate (g) as a raw material.

(iv) Synthesis of 3-amino-6-(2,6-difluorophenyl)-5-fluoro-2-picolinic acid (D2)

Methyl 3-amino-6-bromo-5-fluoro-2-picolinic acid (g) (56 mg, 0.20 mmol) was dissolved in 1:1 tetrahydrofuran/methanol mixed solvent (1 ml), and 1 M lithium hydroxide solution was added. (0.4ml, 0.40mmol), stirred at room temperature for 2 hours, then neutralized with 1N hydrochloric acid to pH=7 and then extracted with ethyl acetate (3×5ml). The extract was washed with water (5ml) and brine (5ml) The aqueous sodium sulfate was dried, filtered and concentrated to give the product (D2) (48mg, 0.18mm).

3. Synthesis of 3-amino-5-fluoro-6-phenyl-2-picolinic acid (D3)

Referring to the synthesis of D2, wherein 2,6-difluorophenylboronic acid (b) is replaced by phenylboronic acid (b1) to obtain a product (D3)

4. Synthesis of 3-fluoro-(2,4'-pyridine)-6-carboxylic acid (D4)

Referring to the synthesis of D1, wherein 2,6-difluorophenylboronic acid (b) is replaced by 4-pyridylboronic acid (b2) to obtain a product (D4)

5. Synthesis of 6-(2,6-difluorophenyl)-3-fluoro-2-picolinic acid (D5)

Referring to the synthesis of D1, wherein 2-bromo-3-fluoro-6-methylpyridine (a) is replaced by 6-bromo-3-fluoro-2-methylpyridine (k) to obtain a product (D5)

6. Synthesis of 6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D6)

(i) 2-methoxycarbonyl-5-trifluoromethyl-pyridine-1-oxide (n)

Methyl 5-trifluoromethyl-2-pyridinecarboxylate (m) (1.03 g, 5.0 mmol) and m-chloroperoxybenzoic acid (1.2 g, 7.0 mmol) dissolved in dichloromethane (15 ml) at 60 ° C After stirring for 6 hours, the solvent was removed and purified by silica gel column chromatography.

(ii) Synthesis of methyl 2-bromo-3-trifluoromethyl-2-pyridinecarboxylate (o)

2-methoxycarbonyl-5-trifluoromethyl-pyridine-1-oxide (n) (221 mg, 1.0 mmol) was added to POBr3 (2.5 g, 8.5 mmol), stirred at 80 ° C for 2 hours and then cooled to It was poured into 20 ml of water at room temperature, and the mixture was extracted with ethyl acetate (3×10 ml). The organic phase was washed with aq. EtOAc (EtOAc) (EtOAc (EtOAc) .

(iii) Synthesis of 6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D6)

Referring to the synthesis of D3, methyl 3-amino-6-bromo-5-fluoro-2-picolinate (g) with methyl 2-bromo-3-trifluoromethyl-2-pyridinecarboxylate (o) Replace the worthy product (D6)

7. Synthesis of 3-amino-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D7)

(i) Synthesis of 2-bromo-3-nitro-5-trifluoromethylpyridine (r)

Was added POBr ₃ (8.6g, 30mmol) in a solution of 3-nitro-5-trifluoromethyl-acetonitrile (25ml) pyridin-2-ol (q) (3.12g, 15mmol) in. After heating to reflux for 4 hours, it was stirred at room temperature for 12 hours. The reaction solution was poured into a stirred aqueous solution of sodium hydrogen carbonate (11 g) (60 ml). The mixture was extracted with dichloromethane (3×20 mL). EtOAcjjjjjjjjjj

(ii) Synthesis of 2-cyano-3-nitro-5-trifluoromethylpyridine (s)

Add tetrabutylammonium bromide (2.2 g, 7.37 mmol) and cyanide to a solution of 2-bromo-3-nitro-5-trifluoromethylpyridine (r) (2.0 g, 7.37 mmol) in toluene (50 ml) Cuprous (CuCN) (1.98 g, 22.2 mmol) was heated under reflux for 9 hours. After cooling to room temperature, water (150 ml) and ethyl acetate (150 ml) were added. The organic phase was washed with water (2×50 mL)

(iii) Synthesis of 3-amino-2-cyano-5-trifluoromethylpyridine (t)

2-Cyano-3-nitro-5-trifluoromethylpyridine (s) (1.0 g, 4.61 mmol) was dissolved in ethyl acetate (20 mL), then 10% palladium / activated carbon (0.10 g) After stirring under hydrogen for 18 hours, it was filtered and concentrated to give a crude product (t) which was directly used for the next reaction.

(iv) Synthesis of 3-amino-5-trifluoromethyl-2-pyridinecarboxylic acid methyl ester (u)

The above 3-amino-2-cyano-5-trifluoromethylpyridine (t) was dissolved in a mixed solvent of concentrated hydrochloric acid (5 ml) and methanol (5 ml). Column purification (5:1 petroleum ether / ethyl acetate) gave product (u) (256 mg, 1.16 mmol).

(v) Synthesis of 3-amino-6-bromo-5-trifluoromethyl-2-pyridinecarboxylic acid methyl ester (v),

3-Amino-5-trifluoromethyl-2-pyridinecarboxylic acid methyl (u) (1.0 g, 4.55 mmol) in water (30 ml), added concentrated sulfuric acid (0.5 ml, 9.1 mmol) and then added bromine (0.23) Ml, 4.55 mmol) in acetic acid (3 mL) (10 min). The reaction was stirred at room temperature for 24 hours, then diluted with H.sub.2 (50 mL). After washing with water (30 ml) and a saturated sodium chloride solution (20 ml), dried over anhydrous sodium sulfate and evaporated. (843 mg, 2.82 mmol).

Synthesis of (vi) 3-amino-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D7)

Referring to the synthesis of D2, 3-amino-6-(2,6-difluorophenyl)-5-fluoro-2-picolinic acid methyl ester with 3-amino-6-bromo-5-trifluoromethyl The product (D7) was prepared by substituting methyl-2-pyridinecarboxylic acid methyl ester (v).

Example 1

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (1)

(1) Preparation of 4-(((5-aminopyrimidin-4-yl)oxy)methyl)piperidine-1-carboxylic acid tert-butyl ester (C1)

Add NaH (67 mg, 2.8 mmol) to a solution of 4-hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ester (B1) (500 mg, 2.32 mmol) in THF (tetrahydrofuran) (10 mL) at room temperature (25 ° C) After stirring for 1 hour, 4-chloropyrimidin-5-amine (H) (346 mg, 2.67 mmol) was added. The reaction was heated to 100 ° C under a nitrogen atmosphere, stirred for 4 hours and concentrated under vacuum at room temperature (20-30 ° C). The residue was purified by silica gel chromatography (EtOAc:EtOAc:EtOAc:EtOAc tert-Butyl-1-carboxylate (C1) (370 mg, 1.2 mmol).

(2) 4-(((5-(6-(2,6-Difluorophenyl)-5-fluoro-2-pyridinecarboxamido)pyrimidin-4-yl)oxy)methyl)piperidine- Preparation of 1-butylic acid tert-butyl ester (E1)

Compound (C1) (49 mg, 0.16 mmol), Compound 6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxylic acid (D1) (40 mg, 0.17 mmol), HATU (77 mg, 0.20 mmol) A mixture of DIEA (88, 0.50 mmol) in DMF (5 mL) was stirred at 50 ° C for one hour. After cooling, it was diluted with ethyl acetate (50 mL) and brine. The organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo at room temperature rotation (20-30 ℃). After concentration, the residue was purified by silica gel column chromatography (eluent: 10-30% ethyl acetate / petroleum ether) to afford product 4-(((5-(6-(2,6-difluorophenyl))- 5-Fluoropyridineamino)pyrimidin-4-yl)oxy)methyl)piperidine-1-carboxylic acid tert-butyl ester (E1) 30 mg, 0.0596 mmol).

(3) Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (1)

At room temperature (25 ℃) conditions, TFA (trifluoroacetic acid) (0.5 mL) was added the compound (E1) (20mg, 0.0394mmol) in CH ₂ Cl ₂ (1mL) was stirred for 10 minutes at room temperature (25 ℃ after) and concentrated under rotary vacuum, the residue was dissolved in CH ₂ Cl ₂ (10mL), a solution of 1 equivalent of sodium hydroxide were used (5mL) and brine (5mL), dried the organic phase over Na ₂ SO ₄ The product was concentrated under vacuum at room temperature (25 ° C) to give the product 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidine-5- Pyridylamide (1) (12 mg, 0.025 mmol).

Example 2

Preparation of N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide (2 )

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxymethyl-azetidin-1-carboxylic acid tert-butyl ester (B2) to give the product 2.

Example 3

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide (3)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (B3) to give the product 3.

Example 4

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide (4)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (B4) to give the product 4.

Example 5

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide (5)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxypiperidine-1-carboxylic acid tert-butyl ester (B5) to give the product 5.

Example 6

Preparation of N-(4-(azetidin-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide (6)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-hydroxy-azetidin-1-carboxylic acid tert-butyl ester (B6) to give the product 6.

Example 7

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (7)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-hydroxymethyl-piperidine-1-carboxylic acid tert-butyl ester (B7) to give the product 7.

Example 8

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (8)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxymethyl-pyrrolidine-1-carboxylic acid tert-butyl ester (B8) to give the product 8.

Example 9

Preparation of N-(4-(azetidin-3-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide (9)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with tert-butyl 3-hydroxy-1-azetidincarboxylate (B9) to give the product 9.

Example 10

Preparation of N-(4-(((r))))) Formamide (10)

The preparation method is referred to in Example 1. Wherein compound B1 of step (1) is replaced with trans-4-hydroxy-cyclohexyl-carbamic acid tert-butyl ester (B10) to give product 10.

Example 11

Preparation of N-(4-((3-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide (11)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxy-cyclohexyl-carbamic acid tert-butyl ester (B11) to give the product 11.

Example 12

Preparation of N-(4-(3-aminopropoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide (12)

The preparation method is referred to in Example 1. Wherein compound B1 of step (1) is replaced with 3-hydroxypropylamine-1-carboxylic acid tert-butyl ester (B12) to give product 12.

Example 13

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(3-(methylamino)propoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (13)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with N-methyl-3-hydroxypropylamine-1-carboxylic acid tert-butyl ester (B13) to give the product 13.

Example 14

Preparation of 6-(2,6-difluorophenyl)-N-(4-(3-(dimethylamino)propoxy)pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide ( 14)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with N,N-dimethyl-3-hydroxypropylamine (B14), and the deprotection of the step (3) is omitted to obtain the product 14.

Example 15

Preparation of N-(4-(4-aminobutoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide (15)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-hydroxybutylamine-1-carboxylic acid tert-butyl ester (B15) to give the product 15.

Example 16

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(4-(methylamino)butoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (16)

The preparation method is referred to in Example 1. Wherein compound B1 of step (1) is replaced with N-methyl-4-hydroxybutylamine-1-carboxylic acid tert-butyl ester (B16) to give product 16.

Example 17

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(3-hydroxypropoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (17)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 1,3-propanediol (B17), and the deprotection of the step (3) is omitted to obtain the product 17.

Example 18

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(3-hydroxybutoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (18)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 1,3-butanediol (B18), and the deprotection of the step (3) is omitted to obtain the product 18.

Example 19

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(4-hydroxybutoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (19)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 1,4-butanediol (B19), and the deprotection of the step (3) is omitted to obtain the product 19.

Example 20

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-((4-hydroxy-4-methylpentyloxy)pyrimidin-5-yl)-2-pyridinecarboxamide ( 20)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-methyl-1,4-pentanediol (B20), and the deprotection of the step (3) is omitted to obtain the product 20.

Example 21

Preparation of 6-(2,6-difluorophenyl)-N-(4-(3,4-dihydroxybutoxy)pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide (21)

(1) 6-(2,6-Difluorophenyl)-N-(4-(2-(2,2-dimethyl-1,3-dioxolan-4-)ethoxy-) Preparation of pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide (E21)

The preparation method is as follows. The method of the steps (1) and (2) in the embodiment 1, wherein the compound B1 of the step (1) is (2,2-dimethyl-1,3-dioxocyclopentan-4-yl) Substituting ethanol (B21) to produce product E21.

(2) 6-(2,6-Difluorophenyl)-N-(4-(3,4-dihydroxybutoxy)pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide (21 Preparation

E21 (20 mg, 0.042 mmol) was dissolved in methanol (2 mL) at room temperature (25 ° C), then concentrated hydrochloric acid (0.5 mL) was added, and stirred for 4 hours, then neutralized with a 10% Na ₂ CO ₃ solution to pH = 7 After adding water (20 mL), vacuum filtration and air drying at 25 ° C gave product 21 (11 mg, 0.025 mmol).

Example 22

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-[4-(tetrahydropyran-4-methoxy)pyrimidin-5-yl)-2-pyridinecarboxamide (22)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with tetrahydropyran-4-methanol (B22), and the deprotection of the step (3) is omitted to obtain the product 22.

Example 23

Preparation of 3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidin-5-yl)-2-pyridine A Amide (23)

The preparation method is referred to in Example 1. Wherein the compound D1 of the step (2) is replaced with 3-amino-6-(2,6-difluoro-phenyl)-5-fluoro-2-pyridinecarboxylic acid (D2) to give the product 23.

Example 24

Preparation of 3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide (twenty four)

The preparation method is referred to Example 23. Wherein compound B1 of step (1) is replaced with B4 to give product 24.

Example 25

Preparation of 3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide (25)

The preparation method is referred to Example 23. Wherein the compound B1 of the step (1) is replaced with B3 to obtain the product 25.

Example 26

Preparation of 3-amino-N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Formamide (26)

The preparation method is referred to Example 23. Wherein compound B1 of step (1) is replaced with B6 to give product 26.

Example 27

Preparation of 3-amino-N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5- Fluor-2-pyridinecarboxamide (27)

The preparation method is referred to Example 23. Wherein compound B1 of step (1) is replaced with B10 to give product 27.

Example 28

Preparation of 3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-ylmethoxy)pyrimidin-5-yl-2-pyridinecarboxamide (28)

The preparation method is referred to Example 23. Wherein compound B1 of step (1) is replaced with B8 to give product 28.

Example 29

Preparation of N-(4-((1-amino-3-chloropropyl-2-yl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro- 2-pyridinecarboxamide (29)

Compound 9 (20 mg, 0.050 mmol) was dissolved in EtOAc EtOAc (EtOAc)

Example 30

Preparation of N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Formamide (30)

The preparation method is referred to Example 29. Compound 9 was replaced with 2 to give product 30.

Example 31

Preparation of 3-amino-N-(4-(azetidin-3-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Formamide (31)

The preparation method is referred to Example 23. Wherein the compound B1 of the step (1) is replaced with B9 to obtain the product 31.

Example 32

Preparation of 3-amino-N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2- Pyridinecarboxamide (32)

The preparation method is referred to Example 23. Wherein compound B1 of step (1) is replaced with B2 to give product 32.

Example 33

Preparation of 3-amino-N-(4-((1-amino-3-chloropropyl-2-yl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl) -5-fluoro-2-pyridinecarboxamide (29)

The preparation method is referred to Example 29. Compound 9 was replaced with compound 31 to give product 33.

Example 34

Preparation of 3-amino-N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5- Fluor-2-pyridinecarboxamide (34)

The preparation method is referred to Example 29. Compound 9 was replaced with compound 32 to give product 34.

Example 35

Preparation of 3-amino-5-fluoro-6-phenyl-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide (35)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with B4, and the compound D1 of the step (2) is replaced with 3-amino-5-fluoro-6-phenyl)-2-pyridinecarboxylic acid (D3) to give the product 35.

Example 36

Preparation of 3-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-(2,4'-bipyridyl)-6-carboxamide (36)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with B3, and the compound D1 of the step (2) is replaced with 3-fluoro-(2,4'-pyridine)-6-carboxylic acid (D4) to obtain the product 36.

Example 37

Preparation of N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-3-fluoro-2-pyridinecarboxamide (37)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with B6, and the compound D1 of the step (2) is replaced with 6-(2,6-difluoro-phenyl)-3-fluoropyridinecarboxylic acid (D5) to obtain the product 37.

Example 38

Preparation of N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)-2-pyridinecarboxamide (38)

Compound 6 (10 mg, 0.023 mmol) was dissolved in 0.1 M sodium hydroxide in methanol (1 mL) and stirred at 50 ° C for 2 hr. The solid was removed after washing with EtOAcqqqqqqqqq

Example 39

Preparation of N-(4-(azetidin-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxamide (39)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-hydroxy-azetidin-1-carboxylic acid tert-butyl ester (B6), and the compound D1 of the step (2) is 6-(2,6-difluorophenyl). Substituting -5-trifluoromethyl-2-picolinic acid (D6) gave product 39.

Example 40

Preparation of 6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidine-5-trifluoromethyl-2-pyridinecarboxamide (40 )

The preparation method is referred to in Example 1. Wherein the compound D1 of the step (2) is replaced with 6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D6) to give the product 40.

Example 41

Preparation of N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridine Amide (41)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxymethyl-azetidine-1-carboxylic acid tert-butyl ester (B2), and the compound D1 of the step (2) is used for 6-(2,6-difluorobenzene). Substituting -5-trifluoromethyl-2-picolinic acid (D6) to give the product 41.

Example 42

Preparation of 6-(2,6-difluorophenyl)-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridinecarboxamide (42 )

The preparation method is referred to in Example 1. Wherein compound B1 of step (1) is replaced with 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (B3), and compound D1 of step (2) is treated with 6-(2,6-difluorophenyl)-5- Instead of trifluoromethyl-2-picolinic acid (D6), product 42 was obtained.

Example 43

Preparation of N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl- 2-pyridinecarboxamide (43)

The preparation method is referred to in Example 1. Wherein compound B1 of step (1) is replaced with t-butyl-cyclohexyl-carbamic acid tert-butyl ester (B10), and compound D1 of step (2) is treated with 6-(2,6-difluorophenyl)-5. Substituting trifluoromethyl-2-picolinic acid (D6) to give the product 43.

Example 44

Preparation of 6-(2,6-difluorophenyl)-N-[4-(tetrahydropyran-4-yl)methoxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridine Formamide (44)

The preparation method is referred to in Example 1. Wherein compound B1 of step (1) is replaced by tetrahydropyran-4-methanol (B22), and compound D1 of step (2) is treated with 6-(2,6-difluorophenyl)-5-trifluoromethyl- Substituting 2-picolinic acid (D6), the deprotection of step (3) is omitted, and product 44 is obtained.

Example 45

Preparation of 3-amino-N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoropyridineamide (45 )

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-hydroxy-azetidin-1-carboxylic acid tert-butyl ester (B6), and the compound D1 of the step (2) is 3-amino-6-(2,6- Substituting difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D7) to give the product 45.

Example 46

Preparation of 3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidine-5-trifluoromethyl-2- Pyridinecarboxamide (46)

The preparation method is referred to in Example 1. Wherein the compound D1 of the step (2) is replaced with 3-amino-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D7) to give the product 46.

Example 47

Preparation of 3-amino-N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl -2-pyridinecarboxamide (47)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxymethyl-azetidine-1-carboxylic acid tert-butyl ester (B2), and the compound D1 of the step (2) is 3-amino-6-(2, Substituting 6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxylic acid (D7) gave product 47.

Example 48

Preparation of 3-amino-6-(2,6-difluorophenyl)-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2- Pyridinecarboxamide (48)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (B3), and the compound D1 of the step (2) is 3-amino-6-(2,6-difluorobenzene). Substituting -5-trifluoromethyl-2-picolinic acid (D7) afforded product 48.

Example 49

Preparation of 3-amino-N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5- Trifluoromethyl-2-pyridinecarboxamide (49)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with trans-4-hydroxy-cyclohexyl-carbamic acid tert-butyl ester (B10), and the compound D1 of the step (2) is 3-amino-6-(2,6-difluoro Substituting phenyl)-5-trifluoromethyl-2-picolinic acid (D7) gave product 49.

Example 50

Preparation of 3-amino-6-phenyl-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridinecarboxamide (50)

The preparation method is referred to in Example 1. Wherein the compound B1 of the step (1) is replaced with 3-hydroxy-pyrrolidine-1-carboxylic acid tert-butyl ester (B4), and the compound D1 of the step (2) is 3-amino-6-(2,6-difluorobenzene). Substituting 5-aminotrifluoromethyl-2-pyridinecarboxylic acid (D7) to give the product 50.

Example 51

Preparation of N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl- 2-pyridinecarboxamide (51)

The preparation method is referred to Example 29. Compound 9 was replaced with compound 41 to give product 51.

Example 52

Preparation of 3-amino-N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5- Trifluoromethyl-2-pyridinecarboxamide (52)

The preparation method is referred to Example 29. Compound 9 was replaced with compound 47 to give product 52.

Example 53

Preparation of N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Formamide (53)

Compound 2 (40 mg, 0.096 mmol) was dissolved in 1:1 trifluoroacetic acid and methanol (1 ml) at room temperature, stirred for 4 hours, then neutralized with saturated sodium carbonate solution, solvent was removed, and the obtained solid was purified by silica gel chromatography. (1:1 ethyl acetate/petroleum ether) gave compound 53 (8 mg, 0.018 mmol)

Example 54

Preparation of 3-amino-N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5- Fluor-2-pyridinecarboxamide (54)

The preparation method is referred to Example 53. Compound 2 was replaced with compound 32 to give product 54.

Example 55

Preparation of N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl- 2-pyridinecarboxamide (55)

The preparation method is referred to Example 53. Compound 2 was replaced with compound 41 to give product 55.

Example 56

Preparation of 3-amino-N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5- Trifluoromethyl-2-pyridinecarboxamide (56)

The preparation method is referred to Example 53. Compound 2 was replaced with compound 47 to give product 56.

Table 1. Structural analysis data of the compounds prepared in the examples of the present invention

Example 57

Effect of R ₃ Substituents on the Inhibition of PIM-1 Kinase Activity by the Compounds of the Invention

In order to verify the effect of the R ₃ substituent on the inhibition of PIM-1 kinase activity by the compounds of the invention (Formula I), we designed 6 groups of compounds with different structures with different functional groups, and synthesized these 6 methods by the methods described in the above examples. Group of compounds. Among the six groups of compounds, the molecular structure of the other moieties in the compound a and the compounds b and c in each group were identical except for the R ₃ substituent. Wherein compound a has R ₃ =H, compound b has R ₃ =F, and compound c has R ₃ =CF ₃ . The difference in their biological activities directly shows the effect of R ₃ on the activity of the compound. This group of compounds 6 to PIM-1 kinase inhibition IC ₅₀ shown in Table 2.

Table 2. Effect of R ₃ substituents on the activity of the compounds of the invention

Data in Table 2 show that all the 5-position-substituted compounds b and c IC ₅₀ than a smaller corresponding compound, such as compound 1a, 1b 3.75 times the IC _50, and thus the active compound is a compound 1a 1b 3.75 Times. Among the six groups of compounds, the activity increase was 2.7 times (compound 4a/4b), and the highest increase was 5.9 times (compound 5a/5b), with an average of 4.2 times. This result indicates that when the R ₃ substituent in the compound of the present invention is F or another substituent, the activity is significantly improved compared to the corresponding compound having no substituent (R ₃ is H), which is related to the existing knowledge. different.

Example 58

Detection and results of the biological activity of the compounds of the invention.

The biological activities of the compounds of the present invention were entrusted to Baonuo Technology (Beijing) Co., Ltd. (E, No. 29, Life Science Park Road, Changping District, Beijing) for testing. The test method is the in vitro activity assay of kinase PIM - IMAP fluorescence polarization method

In vitro activity assay of kinase PIM - IMAP fluorescence polarization method

Principle

PIM is a serine/threonine protein kinase that phosphorylates 5-FAM-labeled small peptide substrates. The unphosphorylated substrate could not bind to the binding reagent (immobilized metal chelate beads) and the fluorescence polarization value was low. The phosphorylated small peptide can be bound to the binding reagent such that the fluorescence polarization value is increased. The degree of phosphorylation of 5-FAM-labeled small peptide substrates reflects the magnitude of PIM kinase activity. The ability of these compounds to inhibit PIM kinase activity can be determined by testing the inhibitory effect of the compounds of the invention on PIM kinase activity at a concentration.

2. Instrument

EnVision, PerkinElmer

3. Reagents and 384-well plates

PIM1 (Millipore Cat. No. 14-573) (purchased from Millipore Corporation, USA)

PIM2 (Millipore Cat. No. 14-607) (purchased from Millipore Corporation, USA)

5-FAM-labeled small peptide (5-FAM-RSRHSSYPAGT, AnaSpec catalog #63801) (purchased from AnaSpec Inc., USA)

IMAP FP Screening Express kit (Molecular Devices catalog #R8127)

(purchased from Molecular Devices, USA)

IMAP Progressive binding reagent (IMAP binder)

IMAP Progressive binding buffer A (5X) (IMAP Binding Buffer A)

IMAP Progressive binding buffer B (5X) (IMAP Binding Buffer B)

384-well black plate (Corning, catalog #3573) (purchased from Corning, USA)

4. Measurement buffer

Tris-HCl (trishydroxymethylaminomethane-hydrochloric acid) (pH 7.2): 10 mM

MgCl ₂ : 10 mM

Triton X-100 (polyethylene glycol octylphenyl ether X-100): 0.01%

DTT (dithiothreitol): 2mM

5. Steps

The compounds of the invention used in this experiment were prepared from the above examples.

a) 10 mM stock of the compound of the invention was diluted to the corresponding concentration with 100% DMSO (dimethyl sulfoxide), and then the compound was diluted 10-fold with a test buffer (dithiothreitol) to give a DMSO concentration of 10%.

b) Measurement system 10ul:

1 ul of compound and 4 ul of enzyme (PIM-1 and PIM-3 final concentration of 0.025 nM, PIM-2 final concentration of 3 nM) were incubated at 23 ° C for 15 minutes, adding 2.5 ul of ATP (PIM-1, PIM-2 and PIM-3) In the activity assay, the final concentrations of ATP were 30 uM, 5 uM and 30 uM, respectively, and 2.5 ul of 5-FAM-labeled small peptide (final concentration of 100 nM) initiated the reaction. The reaction was carried out at 23 ° C for 60 minutes. In the reaction system of the maximum control, DMSO was used instead of the compound, and in the reaction system of the minimum control, the activity buffer (dithiothreitol) was used instead of the enzyme.

c) 30 ul of IMAP binding agent (containing 75% IMAP Binding Buffer A, 25% IMAP Binding Buffer B, 1/600 immobilized metal chelate beads) was added and the reaction was stopped and incubated for 60 minutes at room temperature.

d) The reading plate has a fluorescence polarization value mP, excitation light of 485 nm, and emission light of 530 nm.

6. Data processing

Percent inhibition = (fluorescence polarization value mP - minimum value × 100 / (maximum - minimum value)

PIM kinase activity by biochemical assays to test, all compounds in the examples are embodiments of PIM-1, PIM-2 and PIM-3 kinase activity significantly inhibited, an IC ₅₀ in the range of 0.1-500nM.

It is apparent that the above-described embodiments are merely illustrative of the examples, and are not intended to limit the embodiments. Other variations or modifications of the various forms may be made by those skilled in the art in light of the above description. There is no need and no way to exhaust all of the implementations. Obvious changes or variations resulting therefrom are still within the scope of protection created by this application.

Claims (20)

1. a compound of the formula I, a stereoisomer thereof, a tautomer thereof, and a pharmaceutically acceptable salt thereof

   

   In the above formula I

   R ₁ is -H, -NHR ₄ , halogen (F, Cl, Br, I), -OH, -OC _1-3 hydrocarbyl, -SH, -SC _1-3 hydrocarbyl, C _1-3 hydrocarbyl, halo Base, haloethyl, -CN and -NO ₂ ;

   R ₂ is -H, -NHR ₄ , halogen (F, Cl, Br, I), -OH, -SH, -OC _1-3 hydrocarbon group with or without a substituent, -SC _1-3 hydrocarbon group, C _1-3 hydrocarbyl and C _3-7 cycloalkyl, halomethyl, haloethyl, -CN and -NO ₂ ;

   R ₃ is -NHR ₅ , halogen (F, Cl, Br, I), -OH, -SH, -OC _1-3 hydrocarbon group with or without a substituent, -SC _1-3 hydrocarbon group, C _{1- 3} hydrocarbyl and C _3-7 cycloalkyl, halomethyl, haloethyl, -CN and -NO ₂ ;

   R ₄ is -H, -C(=O)-R ₅ , a C _1-8 hydrocarbon group with or without a substituent, a C _3-7 cycloalkyl group, a 4-7 membered heterocyclic group, 5-10 members Aryl and heterocyclic aryl;

   R ₅ is a C _1-8 hydrocarbon group, a C _1-8 alkoxy group, a C _3-7 cycloalkyl group with or without a substituent;

   R ₆ is a C _3-7 cycloalkyl group, a 4-7 membered heterocyclic group, a 5-10 membered aryl group and a heterocyclic aryl group with or without a substituent; the cyclic hydrocarbon group, an aryl group and a heterocyclic group The substituents on the group may be halogen (F, Cl, Br, I), -CN, -NH ₂ , -NHR ₇ , C _1-4 hydrocarbyl, C _1-4 halohydrocarbyl, C _3-7 cycloalkyl. , -OR ₇ , -NO ₂ , -C(=O)OR ₇ , -C(=O)R ₇ , -C(=O)N(R ₇ ) ₂ , -C(=O)NH ₂ ,- C(=O)NHR ₇ ;

   R ₇ is -H or with or without a substituted C _1-4 hydrocarbyl group;

   R ₂₂ is a C _1-8 hydrocarbon group with or without a substituent or a group defined by the formula:

   

   Wherein R ₂₃ , R ₂₄ and R ₂₅ are each halogen (F, Cl, Br, I), -OR ₁₅ , -NR ₁₆ R ₁₇ , -C(=O)NR ₁₈ R ₁₉ with or without substitution a C _1-8 hydrocarbon group;

   R ₁₅ , R ₁₆ , R ₁₇ , R ₁₈ , R ₁₉ are each -H or a C _1-8 hydrocarbon group with or without a substituent

   G ₁ is CH ₂ or N;

   G ₂ is NR ₂₈ , CHR ₂₉ or O;

   B1 and B2 are 0, 1, 2 or 3;

   B3 is 0, 1, 2;

   B4 is 0,1;

   R ₂₆ and R ₂₇ are each -H or a C _1-8 hydrocarbon group with or without a substituent;

   R ₂₈ is -H, a C _1-8 hydrocarbyl group with or without a substituent, a C _3-7 cycloalkyl group, a C _3-7 heterocycloalkyl group, -C(=O)R ₃₀ , -C(=O ) OR ₃₀ or -C(=O)NHR ₃₀ ;

   R ₂₉ is -H, -OH, a C _1-8 hydrocarbyl group with or without a substituent, a C _3-7 cycloalkyl group, a C _3-7 heterocycloalkyl group, -NHR ₃₀ , -C(=O)OR ₃₀ or -C(=O)NHR ₃₀ ;

   R ₃₀ is -H or a C _1-8 hydrocarbon group with or without a substituent;

   The substituent may be selected from a hydroxyl group, a nitro group, an amino group, an imino group, a cyano group, a halogen group, a thio group, a sulfonyl group, a thioamido group, a thiol group, an anthranylene group, unless otherwise specified. , oxamidino, methoxamidino, sulfhydryl, sulfonylamino, carboxy, formyl, lower alkyl, halogenated lower alkyl, lower alkylamino, halogenated lower alkyl Amino, lower alkoxy, halogenated lower alkoxy, lower alkoxyalkyl, alkylcarbonyl, aminocarbonyl, arylcarbonyl, aralkylcarbonyl, heteroaryl.
2. The compound according to claim 1, wherein

   R ₁ is -H, -NH ₂ , halogen (F, Cl, Br, I), -CN, -NO ₂ or C _1-8 alkoxy;

   R ₂ is -H, halogen (F, Cl, Br, I), -NH ₂ , C _1-8 hydrocarbyl or C _1-8 hydrocarbyloxy;

   R ₃ is halogen (F, Cl, Br, I), a C _1-3 hydrocarbon group with or without a substituent, and a -OC _1-3 hydrocarbon group, a halogenated methyl group, -CN and -NO ₂

   R ₆ is a 5- or 6-membered aryl or heteroaryl group with or without a substituent; the aryl or heteroaryl group may have one or more substituents, each of which is -CN, NO ₂ , halogen (F, Cl, Br, I ), - OH, -NH 2, C 1-4 hydrocarbyloxy, C _1-4 substituted amino, C _1-4 hydrocarbyl or C _3-6 cycloalkyl.
3. The compound according to any one of claims 1 to 2, wherein

   R ₂₂ is cyclopentyl with or without a substituent, cyclohexyl, cycloheptyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, azepanyl, cyclobutylmethyl , cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, azetidinylmethyl, pyrrolidinylmethyl, piperidinylmethyl, piperazinylmethyl, azepane group, butylene group methyl, tetrahydrofuryl methyl or tetrahydropyranyl-methyl; with a substituent group R ₂₂ may have one or more substituents, which may each independently be -NH ₂ , -OH, methyl, ethyl or methoxy.
4. The compound according to any one of claims 1 to 2, wherein

   R ₂₂ is a substituted C ₂ -C ₅ hydrocarbon group, and these hydrocarbon groups may have up to 4 substituents, which may be halogen (F, Cl, Br, I), NH ₂ , methylamino, ethylamine, respectively. , propylamino, dimethylamino, diethylamino, hydroxy, methyl, ethyl, propyl, -CH ₂ OH, -CH ₂ (OH)CH ₃ , -CH ₂ CH ₂ OH, monohalo Base, polyhalomethyl, monohaloethyl, polyhaloethyl, C _1-4 hydrocarbyl-O-, C _1-4 hydrocarbyl-S-.
5. The compound according to any one of claims 1 to 4, wherein R _{3 is} selected from the group consisting of halogen (F, Cl, Br, I), halomethyl, -CN and -NO ₂ .
6. The compound according to any one of claims 1 to 5, wherein R _{3 is} selected from the group consisting of halogen (F, Cl, Br, I) and -CF ₃ .
7. The compound according to any one of claims 1 to 6, wherein R _{6 is} selected from phenyl or pyridyl, which may be selected from one to three -F, -Cl, -Br, -I, - Substituted by a group of OH, -NH _{2 ,} C _1-3 hydrocarbyl, C _1-3 alkoxy, halo C _1-3 hydrocarbyl.
8. The compound according to any one of claims 1 to 7, wherein R ₆ is phenyl, pyridyl, 2-F-6-OCH ₃ -phenyl, 2,6-difluorophenyl.
9. The compound according to claim 1, wherein the compound is the following compound:

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   N-(4-(azetidin-3-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

   N-(4-((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Amide

   N-(4-((3-Aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

   N-(4-(3-Aminopropoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(3-(methylamino)propoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-N-(4-(3-(dimethylamino)propoxy)pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide

   N-(4-(4-Aminobutoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(4-(methylamino)butoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(3-hydroxypropoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-difluorophenyl)-5-fluoro-N-(4-(3-hydroxybutoxy)pyrimidin-5-yl)pyridine amide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(4-hydroxybutoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-((4-hydroxy-4-methylpentyloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-N-(4-(3,4-dihydroxybutoxy)pyrimidin-5-yl)-5-fluoro-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-[4-(tetrahydropyran-4-methoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidin-5-yl)pyridine amide

   3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   3-Amino-N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine A Amide

   3-amino-N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro Pyridylamide

   3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(pyrrolidin-3-ylmethoxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   N-(4-((1-Amino-3-chloropropyl-2-yl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2 -pyridinecarboxamide

   N-(4-(3-Amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine A Amide

   3-amino-N-(4-(azetidin-3-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Amide

   3-amino-N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Formamide

   3-amino-N-(4-((1-amino-3-chloropropyl-2-yl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)- 5-fluoro-2-pyridinecarboxamide

   3-amino-N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro -2-pyridinecarboxamide

   3-amino-5-fluoro-6-phenyl-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-2-pyridinecarboxamide

   3-fluoro-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-(2,4'-bipyridyl)-6-carboxamide

   N-(4-(azetidin-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-3-fluoro-2-pyridinecarboxamide

   N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-5-fluoro-6-(2-fluoro-6-methoxyphenyl)-2-pyridinecarboxamide

   N-(4-(azetidin-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidine-5-trifluoromethyl-2-pyridinecarboxamide

   N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2-pyridinecarboxamide

   6-(2,6-Difluorophenyl)-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridinecarboxamide

   N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2 -pyridinecarboxamide

   6-(2,6-Difluorophenyl)-N-[4-(tetrahydropyran-4-yl)methoxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridine Amide

   3-amino-N-(4-(azepan-4-yloxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoropicolinamide

   3-amino-6-(2,6-difluorophenyl)-5-fluoro-N-(4-(piperidin-4-ylmethoxy)pyrimidine-5-trifluoromethyl-2-pyridine Formamide

   3-Amino-N-(4-(azetidin-3-ylmethoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl- 2-pyridinecarboxamide

   3-amino-6-(2,6-difluorophenyl)-N-(4-(piperidin-4-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridine Formamide

   3-Amino-N-(4-(((1r,4r)-4-aminocyclohexyl)oxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-III Fluoromethyl-2-pyridinecarboxamide

   3-amino-6-phenyl-N-(4-(pyrrolidin-3-yloxy)pyrimidin-5-yl)-5-trifluoromethyl-2-pyridinecarboxamide

   N-(4-(3-Amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2 -pyridinecarboxamide

   3-amino-N-(4-(3-amino-2-(chloromethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-III Fluoromethyl-2-pyridinecarboxamide

   N-(4-(3-Amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro-2-pyridine Amide

   3-amino-N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-fluoro -2-pyridinecarboxamide

   N-(4-(3-Amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-trifluoromethyl-2 -pyridinecarboxamide

   3-amino-N-(4-(3-amino-2-(hydroxymethyl)propoxy)pyrimidin-5-yl)-6-(2,6-difluorophenyl)-5-three Fluoromethyl-2-pyridinecarboxamide.
10. A method of preparing a compound according to any one of claims 1-9, comprising the steps of:

    
11. A method of preparing a compound according to any one of claims 1-9, comprising the steps of:

    The alcohol B with or without a protecting group is first reacted with a base such as NaH in a solvent such as tetrahydrofuran at room temperature for 1 hour, and then reacted with 5-amino-4-chloropyrimidine A under heating. Aminopyrimidine ether C is obtained in 1-10 hours; aromatic carboxylic acid D with or without a protecting group is reacted with amine C in a solvent in the presence of a coupling reagent or a base to obtain an ether compound E; if not in E A protecting group, wherein E is an ether compound of the formula I wherein the substituent E is a hydrocarbyloxy group; if a protecting group such as t-butoxyformate BOC or trimethylsilane, E is trifluoroacetic acid After mixing with dichloromethane or a methanolic solution of hydrochloric acid at room temperature, the mixture is stirred for 1 to 16 hours, and then the solvent is distilled off under reduced pressure at room temperature to obtain an ether compound of the formula I wherein the substituent E is a hydrocarbyloxy group.
12. A method of preparing a compound according to any one of claims 1-9, comprising the steps of:

    1.1 equivalents of alcohol B with or without a protecting group are first reacted with a base such as 1.1 equivalents of NaH in a solvent such as tetrahydrofuran at room temperature for 1 hour and then with 1 equivalent of 5-amino-4-chloro Pyrimidine A can be reacted under heating conditions, for example, at 100 ° C for 1-10 hours to obtain aminopyrimidine ether C; 1 equivalent of aromatic carboxylic acid D with or without a protecting group in a coupling reagent, such as 1.1 equivalents of HATU, base such as 3 In the presence of equivalent DIEA, it is reacted with 1 equivalent of amine C in a solvent such as DMF to obtain an ether compound E; if there is no protecting group in E, E is an ether compound of formula I wherein the substituent E is a hydrocarbyloxy group. If a protective group such as t-butoxyformate BOC or trimethylsilane is present, E is further mixed with 10 to 100 equivalents of trifluoroacetic acid and an equal volume of methylene chloride or 2 equivalents of methanolic hydrochloric acid. After the solution is mixed at room temperature, it is stirred for 1 to 16 hours, and then the solvent is distilled off under reduced pressure at room temperature to obtain an ether compound of the formula I wherein the E substituent is a hydrocarbyloxy group.
13. A method of using the compound of any of claims 1-9, comprising administering an effective amount of a compound of any of claims 1-9 to a subject in need of treatment.
14. Use of a compound according to any one of claims 1-9, characterized in that it is used as a PIM kinase inhibitor in pharmaceuticals.
15. Use of a compound according to claim 14 for use as a pharmaceutical agent for PIM-1, PIM-2 and PIM-3 kinase inhibitors.
16. A method of using the compound of any of claims 1-9, comprising administering an effective amount of a compound according to any one of claims 1-9 to a subject in need of treatment, characterized in that:

    For the treatment or prevention of cancer, reversing the resistance of anticancer and other drugs, inflammatory bowel disease, autoimmune diseases, allergic diseases, atherosclerosis, anti-organ transplant rejection, multi-directional drug resistance, T Cell-mediated inflammation; "cancer" refers to a cancerous disease that can be beneficially treated by inhibiting PIM kinase, including, for example, solid tumors such as cancers such as lung cancer, pancreatic cancer, thyroid cancer, ovarian cancer, bladder cancer, breast cancer, prostate Cancer or colon cancer, melanoma, bone marrow disorders such as myeloid leukemia, multiple myeloma and erythroleukemia, adenomas, for example, villous colon adenomas and sarcomas such as osteosarcoma.
17. A method of using the PIM kinase inhibitor of claim 1 or 8 or 9, comprising administering an effective amount of the PIM kinase inhibitor of claim 9 to a subject in need of treatment, wherein:

    For the treatment or prevention of cancer, reversing the resistance of anticancer and other drugs, inflammatory bowel disease, autoimmune diseases, allergic diseases, atherosclerosis, anti-organ transplant rejection, multi-directional drug resistance, T Cell-mediated inflammation; "cancer" refers to a cancerous disease that can be beneficially treated by inhibiting PIM kinase, including, for example, solid tumors such as cancers such as lung cancer, pancreatic cancer, thyroid cancer, ovarian cancer, bladder cancer, breast cancer, prostate Cancer or colon cancer, melanoma, bone marrow disorders such as myeloid leukemia, multiple myeloma and erythroleukemia, adenomas, for example, villous colon adenomas and sarcomas such as osteosarcoma.
18. A pharmaceutical composition according to any one of claims 1-9, characterized in that it comprises a pharmaceutical composition according to any one of claims 1 to 9 as an active ingredient and a pharmaceutically acceptable carrier.
19. A pharmaceutical composition comprising a PIM kinase inhibitor of the compound of claim 9 comprising a PIM kinase inhibitor of the compound of claim 1 or 8 or 9 as a pharmaceutical composition comprising a pharmaceutical carrier.
20. Use of a pharmaceutical composition according to any one of claims 1-9 for the preparation of a medicament for the treatment or prevention of cancer, reversal of anticancer and other drug resistance, inflammatory bowel disease, autoimmunity Disease, allergic reaction disease, atherosclerosis, anti-organ transplant rejection, multi-directional resistance, T cell-mediated inflammation; "cancer" means can pass A cancerous disease that inhibits PIM kinase from beneficial treatment, including, for example, solid tumors such as lung cancer, pancreatic cancer, thyroid cancer, ovarian cancer, bladder cancer, breast cancer, prostate cancer or colon cancer, melanoma, bone marrow disorders such as myeloid Leukemia, multiple myeloma and erythroleukemia, adenomas, for example, villous colon adenomas and sarcomas such as osteosarcoma; liquid tumors such as chronic lymphocytic leukemia, acute lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, non- Hodgkin's lymphoma, and the application of drugs for multiple myeloma.