CN113135938B - Substituted macrocyclic tyrosine kinase inhibitors and uses thereof - Google Patents

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a substituted macrocyclic tyrosine kinase inhibitor compound, pharmaceutically acceptable salt and stereoisomer thereof. More specifically, the tyrosine kinase is one or more of TRK, ALK and/or ROS1, a pharmaceutical composition and a preparation containing the compound, pharmaceutically acceptable salts and stereoisomers thereof, a method for preparing the compound, the pharmaceutically acceptable salts and the stereoisomers thereof, and application of the compound, the pharmaceutically acceptable salts, the esters and the stereoisomers thereof.

Description

Substituted macrocyclic tyrosine kinase inhibitors and uses thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a substituted macrocyclic tyrosine kinase inhibitor compound, and pharmaceutically acceptable salt or stereoisomer thereof. The tyrosine kinase is one or more of TRK, ALK and/or ROS 1. The invention further relates to pharmaceutical compositions or formulations containing the compounds, pharmaceutically acceptable salts thereof, or stereoisomers thereof. The compounds can be used for preparing medicines for treating pain, inflammation, cancer, neurodegenerative diseases and autoimmune diseases mediated by one or more tyrosine kinase receptors in TRK, ALK and/or ROS 1.

Background

Cancer, also known as malignant tumor, is a serious threat to human health and life. In 2004, 740 million people died of cancer worldwide. In 2008, the third national cause of death survey in china shows that the cancer death rate in china has increased by about eight years in the past 30 years, and nearly 200 million people dying from cancer every year have very severe situation.

Molecular targeted therapy is a major breakthrough in cancer therapy in recent years. Compared with traditional treatment means such as surgery, radiotherapy and chemotherapy, the molecular targeted therapy opens up a new place for the treatment of cancers by using high specificity and relatively low toxic and side effects, and gradually becomes a standard treatment scheme for patients with advanced cancers. Protein kinases, a key regulator of cell growth, proliferation and survival, are a large area of targeted therapy, and both genetic and epigenetic changes may contribute to the development of cancer.

ALK, an anaplastic lymphoma kinase (anaplastic lymphoma kinase), was named for the first time as found in anaplastic large cell lymphoma AMS3 cell line. EML4 belongs to echinoderm microtubule-associated protein family, and is composed of an N-terminal base region, a hydrophobic echinoderm microtubule-associated protein region and a WD repetitive region. The EML4-ALK fusion gene is reported to be related to the formation of tumors, and the function of an N-terminal base region is the most important. Soda, equal to 2007, first reported that the EML4-ALK gene fused in non-small cell lung cancer (NSCLC) as a result of inversion of the short arm of chromosome 2, with EML4 fused to the intracellular ALK kinase region at the N-terminus. The EML4-ALK fusion sites are diverse, and at least 8 EML4-ALK mutants are formed. ALK mutations have been found in a variety of cancers, including Anaplastic Large Cell Lymphoma (ALCL), non-small cell lung cancer, inflammatory myofibroblast tumors, colorectal cancer, breast cancer, and several others.

ROS1 is also a tyrosine kinase receptor currently of great interest. ROS1 is located in the 6q21 region, and the full-length cDNA contains 44 exons, which encode 2347 amino acids with a molecular weight of 259 kDa. The basic structure of ROS1 consists of the extramembranous region (amino acids 1-1861), the transmembrane region (amino acids 1862-2342), and the intramembranous tyrosine kinase-active region (amino acids 1883-2347). The first proto-oncogene fusion site of ROS1 (FIG-ROS1) is found in glioblastoma, and an intermediate deletion of 240 bases on 6q21 results in the expression of FIG-ROS1 fusion protein, which activates tyrosine kinase activity. ROS1 mutations have also been found in a variety of cancers, including glioblastoma, non-small cell lung cancer, colorectal cancer, breast cancer, and the like.

Trk is a nerve growth factor activated tyrosine kinase family, comprises three subtypes of TrkA, TrkB and TrkC, and is respectively encoded by NTRK1 (neurophic receptor tyrosine kinase 1), NTRK2 and NTRK3 genes. After the Trk kinase is phosphorylated, the Trk kinase can activate downstream signal molecules, thereby playing roles in regulating cell proliferation, differentiation, metabolism, apoptosis and the like. The NTRK gene can be fused with other genes, so that the high expression of Trk kinase or the continuous increase of the Trk kinase activity is caused, and finally, the cancer can be caused. NTRK gene fusion occurs in a variety of adult and childhood solid tumors, including breast cancer, colorectal cancer, non-small cell lung cancer, and various sarcomas.

The three tyrosine kinases have strong homology. The homology of the ROS1 gene and the ALK gene in a tyrosine kinase region sequence is 49 percent, the homology of the two genes in an ATP binding site of a kinase catalytic region is up to 77 percent, the homology of the kinase region sequence of TRK A/B/C is more than 80 percent, and the homology of the TRK A gene, the ROS1 gene and the ALK gene in the tyrosine kinase region sequence is about 40 percent. The marketed ALK inhibitor Crizotinib (Crizotinib) has the inhibitory activity of ROS1 and TRK at the same time, and the TRK inhibitor enrotritinib (Entretinib) also has the inhibitory activity of ALK and ROS 1.

At present, the marketed ALK/ROS1 inhibitor and the marketed NTRK inhibitor declared in 2017 show drug resistance in the process of long-term medication, mainly because the gene mutation causes the change of amino acid sequence in kinase protein, such as mutation sites common in ALK kinase as L1196M, L1152R, G1202R, G1269A, 1151Tins, S1206Y, C1156Y, F1174L and the like, mutation sites common in ROS1 kinase as G2032R, D2033N, S1986F, L2026M, L1951R and the like, and mutation sites common in NTRK kinase as G595R, G623R, G667C, G623E, L686M and the like. Therefore, the development of an anti-tumor drug which has strong drug effect and low toxicity and can solve the problem of drug resistance has very important clinical value and significance.

Poor absorption, distribution, metabolism and/or excretion properties (ADME) are the major cause of failure of clinical trials of candidate drugs in clinical trials. The rapid metabolism of drugs can result in the difficulty of obtaining many drugs that are otherwise effective in treating disease due to their rapid metabolic clearance from the body. Although frequent or high dose administration may solve the problem of rapid clearance of the drug, this method may cause side effects and treatment costs associated with high dose administration. Rapidly metabolized drugs may also expose patients to undesirable toxic or reactive metabolites. Thus, it is important to develop a compound with selective inhibitory activity and better pharmacodynamics/pharmacokinetics.

Disclosure of Invention

The technical scheme of the invention provides a substituted macrocyclic tyrosine kinase inhibitor compound, pharmaceutically acceptable salt or stereoisomer thereof, and a composition and application thereof. The compound has good inhibition activity on one or more tyrosine kinases in TRK, ALK and/or ROS1, and further has good inhibition activity on Trk A and/or Trk B and/or Trk C kinase, high exposure and bioavailability in organisms, and good pharmacodynamics and pharmacokinetic properties.

The technical scheme of the invention is as follows:

in one aspect, the present application provides a compound represented by the following general formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

wherein the content of the first and second substances,

M¹、M²、M³、M⁴and M⁵Each independently selected from C (R)¹) Or N, and at least one of which is N;

X¹、X²each independently selected from-C (R)^a)(R^b)-、-N(R^c) -or-O-;

R^a、R^b、R^c、R^d、R^e、R^feach independently selected from hydrogen, deuterium, halogen, nitro, cyano or optionally substituted by 1-3Q³Substituted with the following groups: c optionally substituted by deuterium_1-4Alkyl, -OR^a’、-NR^a’R^b’、-C(O)R^a’、-C(O)OR^a’、-OC(O)R^a’、-OC(O)OR^a’、-OC(O)NR^a’R^b’、-C(O)NR^a’R^b’、-NR^a’C(O)R^b’、-NR^a’C(O)OR^b’and-NR^a’C(O)NR^a’R^b’；

Each Q³Each independently selected from deuterium, halogen, nitro, cyano or the following group optionally deuterated: hydroxy, amino, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_1-4Alkylamino radical, di (C)_1-4Alkyl) amino, halo C_1-4Alkyl, hydroxy C_1-4Alkyl, amino C_1-4Alkyl, halo C_1-4An alkoxy group;

R^a’and R^b’Each independently selected from hydrogen, deuterium, halogen, nitro, cyano or the following optionally deuterated: hydroxy, amino, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_1-4Alkylamino radical, di (C)_1-4Alkyl) amino, halo C_1-4Alkyl, hydroxy C_1-4Alkyl, amino C_1-4Alkyl, halo C_1-4An alkoxy group;

each R¹Each Q¹Each Q²Each independently selected from hydrogen or deuterium;

x is selected from hydrogen, deuterium, halogen, nitro, cyano and optionally deuterated C_1-4An alkyl group;

m is an integer from 0 to 7;

n is an integer from 0 to 2;

and M¹、M²、M³、M⁴、M⁵、X¹、X²Each Q¹Each Q²、X、R^d、R^eAnd R^fAt least one of which is deuterated or is deuterium.

In certain embodiments, the present application provides a compound of the foregoing general formula (I), a pharmaceutically acceptable salt, or a stereoisomer thereof, wherein,

M¹、M²、M³、M⁴and M⁵Each independently selected from C (R)¹) Or N, and at least one of which is N;

X¹is selected from-C (R)^a)(R^b) -or-O-;

X²is selected from-N (R)^c) -or-O-;

R^a、R^b、R^ceach independently selected from hydrogen, deuterium, halogen, nitro, cyano or optionally substituted by 1-3Q³Substituted with the following groups: c optionally substituted by deuterium_1-4Alkyl, -OR^a’、-NR^a’R^b’、-C(O)R^a’、C(O)OR^a’and-C (O) NR^a’R^b’；

Each Q³Each independently selected from deuterium, halogen, nitro, cyano or the following optionally deuterated: hydroxy, amino, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_1-4Alkylamino radical, di (C)_1-4Alkyl) amino, halo C_1-4Alkyl and halo C_1-4An alkoxy group;

R^a’and R^b’Each independently selected from hydrogen, deuterium, halogen, nitro, cyano or the following optionally deuterated: hydroxy, amino, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_1-4Alkylamino radical, di (C)_1-4Alkyl) amino, halo C_1-4Alkyl, hydroxy C_1-4Alkyl, amino C_1-4Alkyl and halo C_1-4An alkoxy group;

R^d、R^e、R^feach independently selected from hydrogen, deuterium or the following optionally deuterated group: c_1-4Alkyl and C_1-4An alkoxy group;

each R¹Each Q¹Each Q²Each independently selected from hydrogen or deuterium;

x is selected from optionally deuterated C_1-4An alkyl group;

m is an integer from 0 to 7;

n is an integer from 0 to 2;

and M¹、M²、M³、M⁴、M⁵、X¹、X²Each Q¹Each Q²、X、R^d、R^eAnd R^fAt least one of which is deuterated or is deuterium.

In certain embodiments, R^a、R^b、R^cEach independently selectsFrom hydrogen, deuterium, fluoro, chloro, bromo, iodo, nitro, cyano, or optionally substituted by 1-3Q³Substituted groups as follows: optionally deuterated C_1-4Alkyl, -OR^a’、-NR^a’R^b’、-C(O)R^a’、-C(O)OR^a’and-C (O) NR^a’R^b’；

Each Q³Each independently selected from deuterium, -OH, -OD, fluoro, chloro, bromo, iodo, nitro, cyano or the following optionally deuterated: amino, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, methoxy, ethoxy, propoxy, isopropoxy, methylamino, dimethylamino, ethylamino, diethylamino, trifluoromethyl and trifluoromethoxy;

R^a’and R^b’Each independently selected from hydrogen, deuterium, -OH, -OD or the following group optionally deuterated: amino, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, methoxy, ethoxy, propoxy, and isopropoxy;

in certain embodiments, R^d、R^e、R^fEach independently selected from hydrogen, deuterium, or the following groups optionally deuterated: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl. In certain embodiments, R^d、R^e、R^fEach independently selected from hydrogen or deuterium.

In certain embodiments, X is selected from the following groups optionally deuterated: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl.

In certain embodiments, X is selected from CH₃、CHD₂、CH₂D and CD₃。

In certain embodiments, m is selected from 0, 1,2, 3,4, 5,6, and 7.

In certain embodiments, n is selected from 0, 1 and 2.

In certain embodiments, M⁵Is N.

In certain embodiments, M²Is C (R)¹)，R¹Selected from hydrogen or deuterium.

In certain embodiments, M⁴Is C (R)¹)，R¹Selected from hydrogen or deuterium.

In certain embodiments, the compounds of formula (I) according to the present invention comprise at least 1 deuterium atom, e.g. comprise 2 deuterium atoms, e.g. comprise 3 deuterium atoms, e.g. comprise 4 deuterium atoms, e.g. comprise 5 deuterium atoms, e.g. comprise 6 deuterium atoms, e.g. comprise 7 deuterium atoms, e.g. comprise 8 deuterium atoms, e.g. comprise 9 deuterium atoms, e.g. comprise 10 deuterium atoms, e.g. comprise 11 deuterium atoms, e.g. comprise 12 deuterium atoms, e.g. comprise 13 deuterium atoms, e.g. comprise 14 deuterium atoms, e.g. comprise 15 deuterium atoms, e.g. comprise 16 deuterium atoms, e.g. comprise 17 deuterium atoms, e.g. comprise 18 deuterium atoms, e.g. comprise 19 deuterium atoms, e.g. comprise 20 deuterium atoms, e.g. comprise 21 deuterium atoms.

In certain preferred embodiments, the deuterium isotope content of deuterium at the deuterium substitution position is at least greater than the natural deuterium isotope content (0.015%), such as greater than 30%, such as greater than 50%, such as greater than 75%, such as greater than 95%, such as greater than 99%.

In certain embodiments, the compound of formula (I), a pharmaceutically acceptable salt or a stereoisomer thereof, described previously herein, is selected from the structures shown below:

wherein X is selected from CH₃、CHD₂、CH₂D or CD₃；

R^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^f、R^15’Each independently selected from hydrogen or deuterium; and when X is selected from CH₃When R is^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^f、R^15’At least one of which is selected from deuterium.

In certain embodiments, X is selected from CH₃、CHD₂、CH₂D or CD₃；

R^1’、R^2’、R^3’、R^11’、R^12’、R^15’Are all hydrogen, R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fEach independently selected from hydrogen or deuterium, and when X is selected from CH₃When R is^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fAt least one of which is selected from deuterium.

In certain embodiments, X is selected from CH₃、CHD₂、CH₂D or CD₃；

R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fAre all hydrogen; r^1’、R^2’、R^3’、R^11’、R^12’、R^15‘Each independently selected from hydrogen or deuterium, and when X is selected from CH₃When R is^1’、R^2’、R^3’、R^11’、R^12’、R^15‘At least one of which is selected from deuterium.

In certain embodiments, the deuterated compound of formula (II), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof described previously herein has a structure according to formula (II-1), formula (II-2), formula (II-3), or formula (II-4) as follows:

wherein, X, R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^d、R^e、R^fAs defined above.

In certain embodiments, X is selected from CH₃、CHD₂、CH₂D or CD₃；

R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^d、R^e、R^fEach independently selected from hydrogen or deuterium, and when X is selected from CH₃When R is^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^d、R^e、R^fAt least one of which is selected from deuterium.

In certain embodiments, X is selected from CH₃；

R^4’、R^5’Are identical and are each selected from a hydrogen atom or a deuterium atom; r^6’、R^7’Are identical and are each selected from a hydrogen atom or a deuterium atom;

R^8’、R^9’are identical and are each selected from a hydrogen atom or a deuterium atom; r^d、R^eAre identical and are each selected from a hydrogen atom or a deuterium atom;

and R is^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^d、R^e、R^fAt least one of which is chosen from deuterium atoms.

In certain embodiments, X is selected from CH₃Or CD₃。

In certain embodiments, X is selected from CH₃，R^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^f、R^15‘Each independently selected from a hydrogen atom or a deuterium atom, and R^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^f、R^15’At least one of which is chosen from deuterium atoms.

In certain embodiments, X is selected from CH₃，R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fAre each a hydrogen atom, R^1’、R^2’、R^3’、R^11’、R^12’、R^15’Each independently selected from a hydrogen atom or a deuterium atom, and R^1’、R^2’、R^3’、R^11’、R^12’、R^15’At least one of which is chosen from deuterium atoms.

In certain embodiments, X is selected from CH₃，R^1’、R^2’、R^3’、R^11’、R^12’、R^15’Are each a hydrogen atom, R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fEach independently selected from a hydrogen atom or a deuterium atom, and R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fAt least one of which is chosen from deuterium atoms.

In certain embodiments, X is selected from CH₃，R^1’、R^2’、R^3’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^f、R^15’Are each a hydrogen atom, R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’Each independently selected from a hydrogen atom or a deuterium atom, and R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’At least one of which is chosen from deuterium atoms.

In certain embodiments, X is selected from CH₃，R^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^15’Are each a hydrogen atom, R^13’、R^14’、R^d、R^e、R^fEach independently selected from a hydrogen atom or a deuterium atom, and R^13’、R^14’、R^d、R^e、R^fAt least one of which is chosen from deuterium atoms.

In certain embodiments, X is selected from CD₃，R^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^f、R^15’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, X is selected from CD₃，R^1’、R^2’、R^3’、R^11’、R^12’、R^15’Are each a hydrogen atom, R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fEach independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, X is selected from CD₃，R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fAre each a hydrogen atom, R^1’、R^2’、R^3’、R^11’、R^12’、R^15’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, X is selected from CD₃，R^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^15’Are each a hydrogen atom, R^13’、R^14’、R^d、R^e、R^fEach independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, X is selected from CD₃，R^1’、R^2’、R^3’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^f、R^15’Are each a hydrogen atom, R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^1’、R^2’、R^3’、R^15’Are respectively provided withIndependently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^1’、R^2’、R^3’、R^15’Are all hydrogen atoms.

In certain embodiments, R^1’、R^2’、R^3’、R^15’Are all deuterium atoms.

In certain embodiments, R^11’、R^12’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^11’、R^12’Are all hydrogen atoms.

In certain embodiments, R^11’、R^12’Are all deuterium atoms.

In certain embodiments, R^4’、R^5’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^4’、R^5’Are the same.

In certain embodiments, R^4’、R^5’Are all hydrogen atoms.

In certain embodiments, R^4’、R^5’Are all deuterium atoms.

In certain embodiments, R^6’、R^7’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^6’、R^7’Are the same.

In certain embodiments, R^6’、R^7’Are all hydrogen atoms.

In certain embodiments, R^6’、R^7’Are all deuterium atoms.

In certain embodiments, R^8’、R^9’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^8’、R^9’Are the same.

In certain embodiments, R^8’、R^9’Are all hydrogen atoms.

In certain embodiments, R^8’、R^9’Are all deuterium atoms.

In certain embodiments, R^13’、R^14’Each independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^13’、R^14’Are the same.

In certain embodiments, R^13’、R^14’Are all hydrogen atoms.

In certain embodiments, R^13’、R^14’Are all deuterium atoms.

In certain embodiments, R^d、R^eEach independently selected from a hydrogen atom or a deuterium atom.

In certain embodiments, R^d、R^eAre the same.

In certain embodiments, R^d、R^eAre all hydrogen atoms.

In certain embodiments, R^d、R^eAre all deuterium atoms.

In certain embodiments, R^10’Is a hydrogen atom.

In certain embodiments, R^10’Is a deuterium atom.

In certain embodiments, R^fIs a hydrogen atom.

In certain embodiments, R^fIs a deuterium atom.

The technical solutions of the present invention can be combined with each other to form a new technical solution, and the formed new technical solution is also included in the scope of the present invention.

In certain embodiments of the present invention, the compounds of the present application, pharmaceutically acceptable salts thereof, and stereoisomers thereof, are selected from the structures shown below:

in an embodiment of the invention, none of the compounds include non-deuterated compounds.

In another aspect, the present application provides a pharmaceutical formulation comprising a compound of the aforementioned general formula ((I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), or formula (VIII), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and one or more pharmaceutically acceptable excipients, which may be in any pharmaceutically acceptable dosage form Emulsifiers, binders, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, ion exchangers, mold release agents, coating agents, flavors, antioxidants, and the like. If necessary, a flavor, a preservative, a sweetener and the like may be further added to the pharmaceutical composition.

In certain embodiments, the pharmaceutical formulations described above may be administered to a patient or subject in need of such treatment by oral, parenteral, rectal, or pulmonary administration, among others. For oral administration, the pharmaceutical composition can be prepared into oral preparations, for example, conventional oral solid preparations such as tablets, capsules, pills, granules and the like; can also be made into oral liquid preparation, such as oral solution, oral suspension, syrup, etc. When the composition is formulated into oral preparations, appropriate filler, binder, disintegrating agent, lubricant, etc. can be added. For parenteral administration, the above pharmaceutical preparations may also be prepared into injections, including injections, sterile powders for injection and concentrated solutions for injection. The injection can be prepared by conventional method in the existing pharmaceutical field, and can be prepared without adding additives or adding suitable additives according to the properties of the medicine. For rectal administration, the pharmaceutical composition may be formulated as a suppository or the like. For pulmonary administration, the pharmaceutical composition may be formulated as an inhalation formulation, aerosol, powder spray, or the like.

In another aspect, the present application also provides a pharmaceutical composition comprising a compound of the aforementioned general formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII) or formula (VIII), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and one or more second therapeutically active agents for use in combination with the tyrosine kinase inhibitor compounds of the present application in the treatment and/or prevention of tyrosine kinase mediated diseases and related conditions, such as pain, cancer, inflammation, neurodegenerative diseases, autoimmune diseases, infectious diseases, and the like.

The pain may be of any origin or etiology, including but not limited to one or more of inflammatory pain, visceral pain, cancer-induced pain, chemotherapy pain, trauma pain, surgical and post-surgical pain, labor pain, acute pain, chronic pain, intractable pain, somatic pain, nociceptive pain, neuropathic pain, blood-borne pain, immunoborne pain, endocrinologically-derived pain, metabolic-induced pain, cardiogenic pain, headache, phantom limb pain, and dental pain. Second therapeutically active agents suitable for use in the combination treatment of pain include, but are not limited to, Nav1.7 channel modulators, opioid analgesics, non-steroidal anti-inflammatory drugs, sedatives, selective/non-selective cyclooxygenase inhibitors, antiepileptics, antidepressants, local anesthetics, 5-HT receptor blockers, 5-HT receptor agonists, ergot alkaloids, beta-receptor blockers, M receptor blockers, nitrates, vitamin K, and the like.

The cancer includes, but is not limited to, one or more of lung cancer, colon cancer, prostate cancer, breast cancer, liver cancer, lymphatic cancer, thyroid cancer, multiple myeloma, soft tissue sarcoma, ovarian cancer, cervical cancer, fallopian tube cancer, renal cell carcinoma, gastric cancer, gastrointestinal stromal tumor, bone cancer, basal cell carcinoma, peritoneal cancer, skin fibroma, pancreatic cancer, esophageal cancer, glioblastoma, head and neck cancer, inflammatory myofibroblastoma, and anaplastic large cell lymphoma. Second therapeutically active agents suitable for use in the combination treatment of cancer include, but are not limited to: mitotic inhibitors, alkylating agents, antimetabolites, antisense DNA or RNA, antitumor antibiotics, growth factor inhibitors, signaling inhibitors, cell cycle inhibitors, enzyme inhibitors, retinoid receptor modulators, proteasome inhibitors, topoisomerase inhibitors, biological response modifiers, hormonal drugs, angiogenesis inhibitors, cytostatic agents, targeting antibodies, HMG-CoA reductase inhibitors, prenyl protein transferase inhibitors, and the like.

The inflammation includes, but is not limited to, atherosclerosis, allergy, and inflammation due to infection or injury. Second therapeutically active agents suitable for use in the combination treatment of inflammation include, but are not limited to: steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs.

The neurodegenerative disease includes, but is not limited to, one or more of alzheimer's disease, parkinson's disease, amyotrophic lateral sclerosis, and huntington's disease. Therapeutically active agents suitable for use in combination therapy for neurodegenerative diseases include, but are not limited to: dopamine-mimetic agents, dopamine receptor agonists, agents affecting dopamine metabolism, NMDA receptor antagonists, adenosine A_2AReceptor inhibitors, drugs that affect DA release and reuptake, central anticholinergics, cholinesterase inhibitors, 5-HT agonists, alpha 2 adrenergic receptor antagonists, antidepressants, cholinergic receptor agonists, beta/gamma secretase inhibitors, H3 receptor antagonists, or anti-oxidant drugs, and the like.

The autoimmune disease includes, but is not limited to, one or more of rheumatoid arthritis, sjogren's syndrome, type I diabetes, and lupus erythematosus. Second therapeutically active agents suitable for use in combination therapy for autoimmune diseases include, but are not limited to: antirheumatic, nonsteroidal anti-inflammatory drug, glucocorticoid drug, TNF antagonist, cyclophosphamide, mycophenolate mofetil, cyclosporine, etc. for improving the disease condition.

Such as trypanosomiasis and the like.

In certain embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients, as described above.

In certain embodiments, the compound of the aforementioned general formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), or formula (VIII), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and the second therapeutically active agent may be present in the same formulation, i.e., in a combination formulation, or may be present in separate formulations for simultaneous or sequential administration to a subject.

In another aspect, the present application also relates to the use of a compound of the aforementioned general formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), or formula (VIII), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the manufacture of a medicament for the treatment and/or prevention of diseases and related conditions mediated by one or more tyrosine kinases of TRK, ALK, and/or ROS 1.

In certain embodiments, the tyrosine kinase-mediated disease and related conditions of one or more of TRK, ALK, and/or ROS1 include, but are not limited to, pain, cancer, inflammation, neurodegenerative disease, autoimmune disease, infectious disease, and the like. Such cancers include, but are not limited to: lung cancer, colon cancer, rectal cancer, prostate cancer, breast cancer, liver cancer, gallbladder cancer, bile duct cancer, leukemia, melanoma, lymphoma, thyroid cancer, multiple myeloma, soft tissue sarcoma, ovarian cancer, cervical cancer, fallopian tube cancer, renal cell carcinoma, stomach cancer, gastrointestinal stromal tumor, bone cancer, basal cell carcinoma, peritoneal cancer, skin fibroma, pancreatic cancer, esophageal cancer, glioblastoma, head and neck cancer, inflammatory myofibroblastoma, anaplastic large cell lymphoma, or neuroblastoma. The pain, inflammation, neurodegenerative disease, autoimmune disease and infectious disease are as defined above.

In certain embodiments, the lung cancer comprises small cell lung cancer and non-small cell lung cancer. In certain embodiments, the non-small cell lung cancer comprises lung adenocarcinoma, squamous cell carcinoma, and large cell carcinoma.

In certain embodiments, the tyrosine kinase-mediated cancer of TRK, ALK, and/or ROS1 comprises a cancer that is at least partially resistant to one or more existing target therapeutic agents.

In certain embodiments, the cancer resistance is caused by one or more mutations in the gene encoding the TRK, ALK, and/or ROS1 kinase receptors.

In certain embodiments, the mutation site of the ALK target is located in L1196M, L1152R, G1202R, G1269A, 1151Tins, S1206A/A6856, F1174A, R1050A, F1245A/L/A1275/A, M1166A, I6851170A, I1171A, V1180A, I1183A, L1196A, a 1200A, L1240A, D1270A, Y1278A, R1192 1124, G A, G1286, T1343A, D1203A, F A/A, F A/A, F A, etc. of the polypeptide. In certain embodiments, the mutation sites of the ROS1 target are located at G2032R, D2033N, S1986F, L2026M, L1951R, L2155S, G2101A, K2003I, etc. of the polypeptide. In certain embodiments, the mutation site of the TRK target is located at G517R, a542V, V573M, F589L, F589C, G595R, G595S, D596V, F600L, F646V, C656Y, C656F, L657V, G667S, G667C, Y676S, G623R, G667C, G623E, L686 685 686M, G545M, a 570M, Q596 685M, V601M, F617M, G623M, D685624 624 4, R630M, C685682, L683 685M, G6853M, G713, C685 4, C685 4, 685 4, G685 4, 685 4, G685 4, 685 4, and G685 4, and G685 4.

In certain embodiments, the compounds of the present application are also useful for treating a condition associated with a disease mediated by a tyrosine kinase selected from JAK2, SRC, FYN, LYN, YES, FGR, FAK, ARK5, or any combination thereof, e.g., cancer.

In another aspect, the present application also provides a method of treating diseases and related conditions mediated by one or more tyrosine kinases of TRK, ALK and/or ROS1, comprising administering to a patient in need thereof an effective amount of a compound of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), or formula (VIII), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, a pharmaceutical formulation thereof, or a pharmaceutical composition thereof; wherein the tyrosine kinase mediated disease and related conditions of one or more of TRK, ALK and/or ROS1 are as described herein before.

By "effective amount" is meant a dosage of a drug that reduces, delays, inhibits or cures a condition in a subject. The size of the administered dose is determined by the administration mode of the drug, the pharmacokinetics of the medicament, the severity of the disease, the individual physical signs of the subject (sex, weight, height, age), and the like.

Detailed Description

In the specification and claims of the present invention, compounds are named according to chemical structure, and if the name and chemical structure of the compound do not match when the same compound is represented, the chemical structure is the standard.

In the present invention, unless otherwise defined, scientific and technical terms used herein have meanings commonly understood by those skilled in the art, however, definitions of some terms are provided below for better understanding of the present invention. Where a definition of a term provided herein does not correspond to a meaning commonly understood by one of ordinary skill in the art, the definition and interpretation of the term provided herein shall control.

The term "halogen" as used herein refers to fluorine, chlorine, bromine or iodine.

"C" according to the invention_1-4Alkyl "denotes straight or branched alkyl having 1 to 4 carbon atoms, including for example" C_1-3Alkyl group "," C_1-2Alkyl group and C_2-3Alkyl group "," C_3-4Alkyl groups "and the like. Examples include, but are not limited to, methyl, ethylAnd (3) a phenyl group, a n-propyl group, an isopropyl group, a n-butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, etc.

"C" according to the invention_1-4Alkoxy "means" C_1-4alkyl-O- ", said" C_1-4Alkyl "is as defined above.

"C" according to the invention_1-4Alkylamino and di (C)_1-4Alkyl) amino "means independently C_1-4alkyl-NH-and

"halo C" as described in this invention_1-4Alkyl group and hydroxyl group C_1-4Alkyl group and amino group C_1-4Alkyl "is independently C_1-4A radical obtained after substitution of the hydrogen of the alkyl group by one or more halogens, hydroxyl groups or amino groups, wherein C is_1-4Alkyl is as defined above.

The "halo C" of the present invention_1-4Alkoxy "means" C_1-4The hydrogen in alkoxy "is substituted with one or more halogens.

The term "optionally substituted with … …" as used herein includes both "substituted with … …" and "unsubstituted with … …".

The term "optionally deuterated" as used herein includes three instances of "unsubstituted", "partially substituted" and "fully substituted" by deuterium.

The "pharmaceutically acceptable salt" of the present invention includes inorganic acid salts, organic acid salts, amino acid salts, organic base salts, or inorganic base salts. Wherein the inorganic acid comprises hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc.; organic acids include acetic acid, trifluoroacetic acid, methanesulfonic acid, toluenesulfonic acid, maleic acid, succinic acid, tartaric acid, citric acid, fumaric acid, and the like; amino acids include arginine, aspartic acid, glutamic acid, and the like; organic bases include meglumine, glucosamine, trimethylamine, triethylamine, dicyclohexylamine, N-dibenzyl-1, 2-ethylenediamine, etc.; the inorganic base includes basic compounds of sodium, potassium, barium, calcium, magnesium, zinc, lithium, etc. These acid or base addition salts can be prepared according to any of the general methods.

"stereoisomers" as used herein refers to compounds of the invention which, when they contain one or more asymmetric centers, may exist as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The compounds of the present invention may have asymmetric centers and thus result in the presence of two optical isomers. The scope of the present invention includes all possible optical isomers and mixtures thereof. The compound of the present invention contains an olefinic double bond, and unless otherwise specified, the scope of the present invention includes cis-isomers and trans-isomers. The compounds of the invention may exist in tautomeric (one of the functional group isomers) forms having different points of attachment of hydrogen through one or more double bond shifts, e.g., a ketone and its enol form are keto-enol tautomers. Each tautomer and mixtures thereof are included within the scope of the present invention. All enantiomers, diastereomers, racemates, meso-isomers, cis-trans-isomers, tautomers, geometric isomers, epimers, mixtures thereof and the like of the compounds are included within the scope of the present invention.

The "dosage form" of the present invention refers to a form prepared from the drug suitable for clinical use, including, but not limited to, powders, tablets, granules, capsules, solutions, emulsions, suspensions, injections (including injections, sterile powders for injections and concentrated solutions for injections), sprays, aerosols, powders, lotions, liniments, ointments, plasters, pastes, patches, gargles or suppositories, more preferably powders, tablets, granules, capsules, solutions, injections, ointments, gargles or suppositories.

The technical solutions cited in the references are included in the disclosure of the present invention, and can be used to explain the contents of the present invention.

Advantageous effects of the invention

1) The compound, the pharmaceutically acceptable salt thereof or the stereoisomer thereof has more excellent tyrosine kinase inhibition activity of one or more of TRK, ALK and/or ROS1, and can treat and/or prevent diseases and related symptoms mediated by one or more tyrosine kinases of TRK, ALK and/or ROS 1.

2) The compound, the pharmaceutically acceptable salt thereof or the stereoisomer thereof improves the metabolism of the compound in organisms, so that the compound has better pharmacokinetic parameter characteristics.

3) The compound, the pharmaceutically acceptable salt thereof or the stereoisomer thereof improves the drug concentration of the compound in animal bodies and improves the drug curative effect.

4) The compound, the pharmaceutically acceptable salt thereof or the stereoisomer thereof improves the safety of the compound.

Detailed description of the preferred embodiments

The technical solutions of the present invention will be described below with reference to examples, and the above-mentioned contents of the present invention will be further described in detail, but it should not be understood that the scope of the present invention is limited only to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Example 1 (2)²R,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1 (5,3) -imidazo [4,5-b]Pyridin-3 (3,2) -pyridin-2 (1,2) -pyrrolidinylcyclooctan-8-one-2²Preparation of (compound 1)

Preparation of 2- (4- (5-fluoro-2-methoxypyridin-3-yl) but-3-yn-1-yl) isoindoline-1, 3-dione

Dissolving 2- (3-yn-1-yl) isoindoline-1, 3-dione (10.0g,50.2mmol), 3-bromo-5-fluoro-2-methoxypyridine (10.3g,50.0mmol) in N, N-dimethylformamide (200mL), adding cuprous iodide (0.95g,5.0mmol), palladium tetratriphenylphosphine (2.8g,2.4mmol), triethylamine (20.2g,200mmol), heating to 90 deg.C, cooling to 25 deg.C, adding methanol (100mL), filtering to obtain a solid, vacuum drying to 12.5g, yield: 77.2 percent.

LC-MS(M/e):325.1(M+H⁺)

Preparation of 4- (5-fluoro-2-methoxypyridin-3-yl) but-3-ynyl-1-amine

2- (4- (5-fluoro-2-methoxypyridin-3-yl) but-3-yn-1-yl) isoindoline-1, 3-dione (12.5g,38.5mmol), dichloromethane/methanol 5:1(150ml) mixed solvent, hydrazine hydrate (13ml) added dropwise, 25 ℃,12 h, filtration, cake washing with dichloromethane (50ml), organic phase washing with sodium chloride solution (50ml), organic phase drying with anhydrous sodium sulfate, filtration, spin drying to give 7g crude product which was used directly in the next reaction.

LC-MS(M/e):195.1(M+H⁺)

Preparation of 3- (3, 4-dihydro-2H-pyrrol-5-yl) -5-fluoro-2-methoxypyridine

4- (5-fluoro-2-methoxypyridin-3-yl) but-3-ynyl-1-amine (7.0g,36.0mmol) was dissolved in acetonitrile (125ml) and water (38ml), palladium chloride (330mg, 1.9mmol) was added, the mixture was heated to 80 ℃ for 16 hours, the solvent was dried, water (100ml) was added, extraction was performed with dichloromethane (3 × 100ml), the organic phases were combined, dried, and the residue was subjected to column chromatography (petroleum ether: ethyl acetate ═ 4:1) to give 4.5g of the compound with a yield of 64.3%.

LC-MS(M/e):195.1(M+H⁺)

Preparation of 5-fluoro-2-methoxy-3- (pyrrolidin-2-yl-2-d) pyridine

3- (3, 4-dihydro-2H-pyrrol-5-yl) -5-fluoro-2-methoxypyridine (3.4g,17.5mmol), D-methanol (10ml), deuterium oxide (2ml), sodium borodeuteride (1.5g, 35.8mmol), 25 ℃,16H, LC-MS detection reaction completion, spin-drying, adding water (50ml), dichloromethane (3X 50ml) extraction, combining organic phases, drying with anhydrous sodium sulfate, filtration, spin-drying to obtain 3.4g crude product, which is used directly in the next step.

LC-MS(M/e):198.1(M+H⁺)

Preparation of 6- (2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl-2-d) -3-nitropyridin-2-amine

5-fluoro-2-methoxy-3- (pyrrolidin-2-yl-2-d) pyridine (3.4g,17.5mmol), 6-chloro-3-nitropyridin-2-amine (2.9g,16.7mmol), acetonitrile (50ml), N-diisopropylethylamine (6.7g,51.9mmol) was added, the mixture was heated to 90 ℃ for 3h, spun dry, water (50ml) was added, filtered and the solid dried under vacuum to give 5.4g of crude product.

LC-MS(M/e):335.1(M+H⁺)

6.6- (2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl-2-d) pyridine-2, 3-diamine preparation

6- (2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl-2-d) -3-nitropyridin-2-amine (3.0g,9.0mmol), methanol (100ml), Raney's nickel (300mg) added, 25 ℃ C., 16h, filtered, spun dry to give 2.5g of crude which was used directly in the next step.

LC-MS(M/e):305.2(M+H⁺)。

Preparation of 5- (2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl-2-d) -3H-imidazo [4,5-b ] pyridine

6- (2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl-2-d) pyridine-2, 3-diamine (2.5g,8.2mmol), trimethyl orthoformate (8.7g,82mmol), toluene (50ml), p-toluenesulfonic acid (140mg,0.81mmol) was added and heated to 110 ℃ for 5h, spun dry, dichloromethane (100ml) was added, sodium bicarbonate solution (30ml) was washed, the organic phase was dried over anhydrous sodium sulfate, filtered, spun dry to give 2.5g crude which was used directly in the next step.

LC-MS(M/e):315.2(M+H⁺)

Preparation of 3- (1- (3H-imidazo [4,5-b ] pyridin-5-yl) pyrrolidin-2-yl-2-d) -5-fluoropyridin-2-ol

5- (2- (5-fluoro-2-methoxypyridin-3-yl) pyrrolidin-1-yl-2-d) -3H-imidazo [4,5-b ] pyridine (2.5g,8.0mmol), hydrogen chloride dioxane solution (40ml), heated to 90 ℃, for 16H, spun dry, triethylamine added, spun dry and preparative purification by medium pressure reverse phase (methanol: water ═ 3:7) afforded 500mg of the compound in yield: 20.8 percent.

LC-MS(M/e):301.1(M+H⁺)

((2R) -1- ((3- (1- (3H-imidazo [4,5-b ] pyridinyl-5-yl) pyrrolidinyl-2-yl-2-d) pyridinyl-2-yl-5-d) oxy) propane

Preparation of (2-yl) carbamic acid tert-butyl ester

3- (1- (3H-imidazo [4,5-b ] pyridin-5-yl) pyrrolidin-2-yl-2-d) -5-fluoropyridin-2-ol (390mg,1.3mmol), (R) - (1-hydroxypropan-2-yl) carbamic acid tert-butyl ester (275mg,1.6mmol), tetrahydrofuran (5ml), azobisformyldipiperidine (656mg,2.6mmol), 0 ℃, addition of tri-n-butylphosphine (526mg,2.6mmol), 25 ℃,16H, spin-drying, addition of water (30ml), dichloromethane (3X 30ml) extraction, combination of the organic phases, spin-drying, column chromatography of the residue (dichloromethane: methanol 30:1) to give 400mg of compound in 67.3% yield.

LC-MS(M/e):458.2(M+H⁺)

Preparation of (2R) -1- ((3- (1- (3H-imidazo [4,5-b ] pyridin-5-yl) pyrrolidin-2-yl-2-d) -5-fluoropyridin-2-yl) oxy) propan-2-amine

Tert-butyl ((2R) -1- ((3- (1- (3H-imidazo [4,5-b ] pyridinyl-5-yl) pyrrolidinyl-2-yl-2-d) pyridinyl-2-yl-5-d) oxy) propan-2-yl) carbamate (0.9g,2.0mmol), hydrogen chloride dioxane solution (50ml), 25 ℃,4 hours, spin-dried and the residue was used directly in the next step.

LC-MS(M/e):358.2(M+H⁺)

11.(2²R,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1 (5,3) -imidazo [4,5-b]Pyridin-3 (3,2) -pyridin-2 (1,2) -pyrrolidinylcyclooctan-8-one-2²Preparation of (d)

(2R) -1- ((3- (1- (3H-imidazo [4,5-b ] pyridin-5-yl) pyrrolidin-2-yl-2-d) -5-fluoropyridin-2-yl) oxy) propan-2-amine (crude from above, 2.0mmol), dichloromethane (10ml), carbonyldiimidazole (650mg,4.0mmol), triethylamine (405mg,4.0mmol), 25 ℃,16H, dichloromethane (20ml), water (10ml) washed, the organic phase dried and the residue purified by column chromatography (petroleum ether: ethyl acetate ═ 1:1) to give 320mg of mixture, purified by reverse phase medium pressure preparation to give 30mg of the compound of the single configuration: 4.0 percent.

Molecular formula C₁₉H₁₈DFN₆O₂Molecular weight 383.4 LC-MS (M/e):384.2(M + H)⁺)

¹H-NMR(400MHz,CDCl₃)δ:10.53(d,J＝7.2Hz,1H),8.38(s,1H),7.80-7.91(m,2H),7.34(d,J＝7.2Hz,1H),6.52(d,J＝9.2Hz,1H),5.15-5.18(m,1H),4.39-4.45(m,1H),4.17-4.23(m,1H),3.93-3.99(m,1H),3.60-3.65(m,1H),2.43-2.58(m,2H),2.23-2.28(m,1H),1.97-2.02(m,1H),1.58(d,J＝6.8Hz,3H)

Example 2 (2)²R,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1 (5,3) -imidazo [4,5-b]Pyridin-3 (3,2) -pyridin-2 (1,2) -pyrrolidinylcyclooctan-8-one-5, 5-d₂Preparation of (Compound 2)

Preparation of methyl ((benzyloxy) carbonyl) -D-alanine

((benzyloxy) carbonyl) -D-alanine (6.0g,26.9mmol), methanol (50ml), thionyl chloride (6.4g,53.8mmol) was added, heated to 90 ℃ for 16h, spun dry, and the residue was subjected to column chromatography (petroleum ether: ethyl acetate 4:1) to give 3.0g of compound (R) - (1-hydroxypropan-2-yl-1, 1-D) in 46.9% yield 2₂) Preparation of benzyl carbamate

Deuterated lithium aluminum hydride (1.05g,25.0mmol) is suspended in tetrahydrofuran (130ml), ((benzyloxy) carbonyl) -D-alanine methyl ester (3.0g,12.6mmol) is added in portions at 0 ℃, stirred for 4 hours at 25 ℃, quenched with sodium sulfate decahydrate solution at 0 ℃, filtered, water (100ml) is added to the filtrate, ethyl acetate (2X 150ml) is extracted, the organic phases are combined, dried over anhydrous sodium sulfate, filtered, spun-dried, and the residue is subjected to column chromatography (petroleum ether: ethyl acetate 1:1) to obtain 2.2g of the compound with the yield: 81.5%.

LC-MS(M/e):212.1(M+H⁺)

Preparation of (R) -3- (1- (6-amino-5-nitropyridin-2-yl) pyrrolidin-2-yl) -5-fluoropyridin-2-ol

(R) -5-fluoro-3- (pyrrolidin-2-yl) pyridin-2-ol hydrochloride (5.0g,21.5mmol), 6-chloro-3-nitropyridin-2-amine (3.9g,22.4mmol), acetonitrile (50ml), N-diisopropylethylamine (8.3g,64.3mmol) was added, the mixture was heated to 90 ℃ for 3 hours, spun dry, water (50ml) was added, the solid was filtered and dried in vacuo to give 6.8g of crude.

LC-MS(M/e):320.1(M+H⁺)

Preparation of (R) -3- (1- (5, 6-diaminopyridin-2-yl) pyrrolidin-2-yl) -5-fluoropyridin-2-ol

(R) -3- (1- (6-amino-5-nitropyridin-2-yl) pyrrolidin-2-yl) -5-fluoropyridin-2-ol (3.0g,9.4mmol), methanol (50ml), palladium on carbon (500mg) added, hydrogen purged at 25 ℃ for 16h, filtered and spun dried to give 2.0g of crude product which was used directly in the next step.

LC-MS(M/e):290.1(M+H⁺)

Preparation of (R) -3- (1- (3H-imidazo [4,5-b ] pyridin-5-yl) pyrrolidin-2-yl) -5-fluoropyridin-2-ol

(R) -3- (1- (5, 6-diaminopyridin-2-yl) pyrrolidin-2-yl) -5-fluoropyridin-2-ol (2.0g,6.9mmol), trimethyl orthoformate (7.3g,69mmol), toluene (50ml), p-toluenesulfonic acid (120mg,0.7mmol) added, heated to 110 ℃,16h, spun dry, sodium bicarbonate solution (30ml) added, dichloromethane (3 × 50ml) extracted, the organic phases combined, spun dry and the residue was purified by medium pressure reverse phase prep (methanol: water ═ 1:1) to give 880mg of compound, yield: 42.5 percent.

LC-MS(M/e):300.1(M+H⁺)

((R) -1- ((3- ((R) -1- (3H-imidazo [4, 5-b))]Pyridyl-5-yl) pyrrolidinyl-2-yl) -5-fluoropyridin-2-ylOxy) prop-2-yl-1, 1-d₂) Preparation of benzyl carbamates

(R) -3- (1- (3H-imidazo [4, 5-b)]Pyridin-5-yl) pyrrolidin-2-yl) -5-fluoropyridin-2-ol (400mg,1.34mmol), (R) - (1-hydroxypropan-2-yl-1, 1-d₂) Benzyl carbamate (340mg,1.61mmol), tetrahydrofuran (10ml), azobisformyldipiperidine (676mg,2.68mmol), 0 ℃, tri-n-butylphosphine (540mg,2.67mmol) was added, 25 ℃,16h, spin-dried, water (30ml) was added, dichloromethane (3 × 30ml) was extracted, the organic phases were combined, spin-dried, and the residue was purified by column chromatography (dichloromethane: methanol 25:1) to yield 250mg of compound, 38.0% yield.

LC-MS(M/e):493.3(M+H⁺)

(R) -1- ((3- ((R) - (1- (3H-imidazo [4, 5-b))]Pyridin-5-yl) pyrrolidinyl-2-yl) -5-fluoropyridin-2-yl) oxy) propane-1, 1-d₂Preparation of (E) -2-amines

((R) -1- ((3- ((R) -1- (3H-imidazo [4, 5-b)]Pyridyl-5-yl) pyrrolidinyl-2-yl) -5-fluoropyridin-2-yl) oxy) prop-2-yl-1, 1-d₂) Benzyl carbamate (250mg,0.5mmol), methanol (10ml), palladium on carbon (100mg) added, hydrogen bubbled at 25 deg.C for 6 hours, filtered, and dried by spin-drying to give 150mg of compound, yield: 82.4 percent.

LC-MS(M/e):359.2(M+H⁺)

8.(2²R,6R)-3⁵-fluoro-6-methyl-1³H-4-oxa-7-aza-1 (5,3) -imidazo [4,5-b]Pyridin-3 (3,2) -pyridin-2 (1,2) -pyrrolidinylcyclooctan-8-one-5, 5-d₂Preparation of

((R)-1- ((3- ((R) -1- (3H-imidazo [4, 5-b)]Pyridyl-5-yl) pyrrolidinyl-2-yl) -5-fluoropyridin-2-yl) oxy) prop-2-yl-1, 1-d₂) Benzyl carbamate (150mg,0.42mmol), dichloromethane (5ml), carbonyldiimidazole (136mg,0.84mmol), triethylamine (85mg,0.84mmol), 25 ℃,16h, dichloromethane (20ml), water (10ml) were added, the organic phase was spun dry, and the residue was purified by column chromatography (petroleum ether: ethyl acetate 1:1) to give 30mg of compound, yield: 18.6 percent.

Molecular formula C₁₉H₁₇D₂FN₆O₂Molecular weight 384.4LC-MS (M/e):385.2(M + H)⁺)

¹H-NMR(400MHz,CDCl₃)δ:10.52(d,J＝7.6Hz,1H),8.38(s,1H),7.87-7.91(m,2H),7.34(d,J＝8.4Hz,1H),6.52(d,J＝8.4Hz,1H),5.63-5.66(m,1H),4.40-4.42(m,1H),3.93-3.98(m,1H),3.60-3.66(m,1H),2.51-2.58(m,2H),2.22-2.28(m,1H),1.97-2.02(m,1H),1.58(d,J＝9.2Hz,3H)

Other table compounds can be prepared by the methods of preparation described in reference to examples 1 and 2, using appropriate starting materials and intermediates.

Activity assay

Exemplary activity tests of some of the compounds of the invention are provided below to show the advantageous activity and beneficial technical effects of the compounds of the invention. It should be understood, however, that the following experimental protocols are only illustrative of the present disclosure and are not intended to limit the scope of the present disclosure.

Experimental example 1 in vitro cytological inhibitory Activity of Compounds of the invention

Test substances the chemical names and structures of some of the compounds of the present invention are shown in the preparation examples.

The cell lines used in the following experiments represent the following:

Ba/F3 SLC34A2/ROS1 cell line:

Ba/F3 cells were transfected with a stably expressing cell line of SLC34A2/ROS 1;

Ba/F3 SLA34A2-ROS1-G2032R cell line:

Ba/F3 cells were transfected with a stably expressing cell line of SLA34A2-ROS 1-G2032R.

Experimental method (CelltiterGlo assay)

1. Preparation of cells

1.1 cell culture:

all cells were suspension cells, the medium was RPMI-1640+ 10% FBS, and the cells were tested in logarithmic growth phase.

1.2 cell suspension preparation:

cells in the logarithmic growth phase were harvested and counted using a platelet counter. Cell viability was checked by trypan blue exclusion to ensure cell viability above 90%. Adjusted to the appropriate concentration, 90 μ L of cell suspension was added to 96-well plates, respectively.

TABLE 1 cell seeding number

2. Formulating test compounds

2.1 test compound DMSO stock solutions were prepared at concentrations given in Table 2.

TABLE 2 stock solution concentration (mM) of test compound

2.2 preparation of working stock solutions of test Compounds

Test compound stock was diluted 10mM to 1mM and then diluted with DMSO in 3-fold serial gradients for a total of 9 concentrations. Then, 2. mu.L of each DMSO-diluted compound was added to 198. mu.L of the culture solution as a working stock solution of the test compound (10-fold concentration of the compound, 1% concentration of DMSO, and 10. mu.M concentration of the highest concentration)

The solvent control wells are DMSO solvent controls, and the blank control wells are filled with medium only, without seeding the cells.

2.3 treatment of Compounds

mu.L of compound working stock (10-fold dilution, final DMSO concentration of 0.1%) was added to each well of a 96-well plate seeded with cells.

The final concentrations of test compounds were: 1000nM, 333.33nM, 111.11nM, 37.04nM, 12.35nM, 4.12nM, 1.37nM, 0.46nM, 0.15 nM.

5％CO₂The cells were cultured in a cell incubator for 72 hours.

3. Detection of

The CTG reagent was thawed and the cell plates were equilibrated to room temperature for 30 minutes, 60. mu.L of reagent (Celltiter Glo assay kit) was added to each well, shaken for 2min with a shaker (protected from light), and incubated at room temperature for 18 minutes (protected from light). And reading the light signal value by a multifunctional microplate reader.

4. Data processing

1) Inhibition (%) × (DMSO solvent control well reading-test well reading)/(DMSO solvent control well reading-blank control well reading) × 100%;

2) inputting GraphPad Prism 5.0 to plot to obtain curves and IC₅₀。

Results and conclusions of the experiment

TABLE 3 in vitro cytological Activity (IC) of Compounds of the invention₅₀,nM)

As can be seen from Table 3, the compound of the invention can effectively inhibit the proliferation of Ba/F3 SLC34A2/ROS1 and Ba/F3 SLA34A2-ROS1-G2032R cells, and shows that the compound of the invention has the clinical application potential of treating ROS1 kinase-mediated cancerous diseases, including drug-resistant cancerous diseases caused by ROS1 gene mutation.

Experimental example 2 in vitro cytological inhibitory Activity of Compounds of the invention

Test substances the chemical names and structures of some of the compounds of the present invention are shown in the preparation examples.

The cell lines used in the following experiments represent the following:

Ba/F3 LMNA-NTRK1-G595R cell line:

Ba/F3 cells were transfected with a stably expressing cell line of LMNA-NTRK 1-G595R;

Ba/F3 ETV6-NTRK3-G623R cell line:

Ba/F3 cells were transfected with a stably expressing cell line of ETV6-NTRK 3-G623R.

Experimental method (Celltiterglo assay)

1. Preparation of cells

1.1 cell culture:

all cells were suspension cells and the medium was RPMI-1640+ 10% FBS, cells were tested in log phase growth.

1.2 cell suspension preparation:

cells in the logarithmic growth phase were harvested and counted using a platelet counter. Cell viability was checked by trypan blue exclusion to ensure cell viability above 90%. Adjusted to the appropriate concentration, 90 μ L of cell suspension was added to 96-well plates, respectively.

TABLE 4 cell seeding number

2. Formulating test compounds

2.1 test compound DMSO stock solutions were prepared at concentrations shown in Table 5.

TABLE 5 stock solution concentration (mM) of test compound

2.2 preparation of working stock solutions of test Compounds

Test compound stock was diluted to 1mM at 10mM and then diluted with DMSO in 3-fold serial gradients for a total of 9 concentrations. Then, 2. mu.L of each DMSO-diluted compound was added to 198. mu.L of the culture solution as a working stock solution of the test compound (10-fold concentration of the compound, 1% concentration of DMSO, and 10. mu.M concentration of the highest concentration)

The solvent control wells are DMSO solvent controls, and the blank control wells are filled with medium only, without seeding the cells.

2.3 treatment of Compounds

mu.L of compound working stock (10-fold dilution, final DMSO concentration of 0.1%) was added to each well of a 96-well plate seeded with cells.

The final concentrations of test compounds were: 1000nM, 333.33nM, 111.11nM, 37.04nM, 12.35nM, 4.12nM, 1.37nM, 0.46nM, 0.15 nM.

5％CO₂The cells were cultured in a cell incubator for 72 hours.

3. Detection

The CTG reagent was thawed and the cell plates were equilibrated to room temperature for 30 minutes, 60. mu.L of reagent (Celltiter Glo assay kit) was added to each well, shaken for 2min with a shaker (protected from light), and incubated at room temperature for 10 minutes (protected from light). And reading the light signal value by a multifunctional microplate reader.

4. Data processing

1) Inhibition (%) × (DMSO solvent control well reading-test well reading)/(DMSO solvent control well reading-blank control well reading) × 100%;

2) inputting GraphPad Prism 5.0 to plot to obtain curves and IC₅₀。

Results and conclusions of the experiment

TABLE 6 in vitro cytological Activity (IC) of Compounds of the invention₅₀,nM)

As shown in Table 6, the compound can effectively inhibit the proliferation of cells such as Ba/F3 LMNA-NTRK1-G595R, Ba/F3 ETV6-NTRK3-G623R and the like, and the compound has the clinical application potential of treating drug-resistant cancerous diseases caused by NTRK gene mutation.

Claims (10)

1. A compound of formula (II), formula (III) or formula (IV), a pharmaceutically acceptable salt thereof or a stereoisomer thereof,

，

wherein X is selected fromCH₃、CHD₂、CH₂D or CD₃；

R^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^fEach independently selected from hydrogen or deuterium; and when X is selected from CH₃When R is^1’、R^2’、R^3’、R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^11’、R^12’、R^13’、R^14’、R^d、R^e、R^fAt least one of which is deuterium.

2. The compound of claim 1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

x is selected from CH₃、CHD₂、CH₂D or CD₃；

R^1’、R^2’、R^3’、R^11’、R^12’Are all hydrogen; r^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fEach independently selected from hydrogen or deuterium; and when X is selected from CH₃When R is^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^13’、R^14’、R^d、R^e、R^fAt least one of which is deuterium.

3. The compound of claim 2, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, having a structure represented by formula (II-1), formula (II-2), formula (II-3), or formula (II-4) below:

wherein the content of the first and second substances,

x is selected from CH₃、CHD₂、CH₂D or CD₃；

R^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^d、R^e、R^fEach independently selected from hydrogen or deuterium, and when X is selected from CH₃When R is^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^d、R^e、R^fAt least one of which is deuterium.

4. The compound of claim 3, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

x is selected from CH₃；

R^4’、R^5’Are identical and are each selected from a hydrogen atom or a deuterium atom; r^6’、R^7’Are identical and are each selected from a hydrogen atom or a deuterium atom; r^8’、R^9’Are identical and are each selected from a hydrogen atom or a deuterium atom; r^d、R^eAre identical and are each selected from a hydrogen atom or a deuterium atom; and R is^4’、R^5’、R^6’、R^7’、R^8’、R^9’、R^10’、R^d、R^e、R^fAt least one of which is a deuterium atom.

5. The compound of claim 1, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, selected from the following compounds:

。

6. a pharmaceutical formulation comprising a compound according to any one of claims 1 to 5, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, and one or more pharmaceutically acceptable carriers and/or excipients.

7. Use of a compound of any one of claims 1-5, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the manufacture of a medicament for the treatment and/or prevention of a disease and related condition mediated by one or more tyrosine kinases of TRK and/or ROS1, said disease and related condition selected from one or more of pain, cancer, inflammation, neurodegenerative disease, autoimmune disease.

8. Use of a compound of claim 7, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, for the manufacture of a medicament for the treatment and/or prevention of diseases and related conditions mediated by one or more tyrosine kinases of TRK and/or ROS1, said diseases being cancers that have developed at least partial resistance to TRK and/or ROS1 therapeutic agents.

9. The use of claim 8, wherein the disease mediated by one or more tyrosine kinases of TRK and/or ROS1 is a cancer selected from the group consisting of lung cancer, colon cancer, rectal cancer, prostate cancer, breast cancer, liver cancer, gall bladder cancer, bile duct cancer, leukemia, melanoma, lymphoma, multiple myeloma, soft tissue sarcoma, ovarian cancer, cervical cancer, fallopian tube cancer, renal cell carcinoma, gastric cancer, gastrointestinal stromal tumors, bone cancer, basal cell carcinoma, peritoneal cancer, pancreatic cancer, esophageal cancer, glioblastoma, head and neck cancer, inflammatory myofibroblastoma, and neuroblastoma.

10. The use of claim 9, wherein the lung cancer is selected from small cell lung cancer and non-small cell lung cancer.