Patent Description:
Receptor Tyrosine Kinase (RTK) is a class of transmembrane enzyme-linked receptor whose overexpression or overactivation is closely associated with the occurrence and development of tumors. Fibroblast Growth Factor Receptors (FGFRs) and the RET protein encoded by oncogene RET (Rearranged during Transfection) are important members of the RTK superfamily and important targets for tumor therapy.

FGFRs mainly include four subtypes, FGFR1, FGFR2, FGFR3 and FGFR4 (<NPL>; <NPL>. ) Overexpression or overactivation of FGFRs by means of gene amplification, mutation, fusion or ligand induction, plays an important role in promoting tumor cell proliferation, invasion, migration and tumor angiogenesis. It is found in the study that FGFRs are overexpressed or overactivated in various tumors such as non-small cell lung cancer, breast cancer, stomach cancer, bladder cancer, endometrial cancer, prostate cancer, cervical cancer, colon cancer, esophageal cancer, keratinoma, myeloma, rhabdomyosarcoma, etc. (<NPL>; <NPL>. ) For example, overactivation of FGFR1 signaling pathway in squamous cell carcinoma of non-small cell lung cancer is up to <NUM>%; (<NPL>),<NPL>);<NPL>. ) The overactivation of FGFR2 signaling pathway in gastric cancer accounts for <NUM>-<NUM>% (<NPL>. FGFR3 mutation in bladder cancer accounts for <NUM>%-<NUM>% (non-invasive) and <NUM>% - <NUM>% (invasive). Various subtypes of FGFR are overexpressed and overactivated in liver cancer, such as FGFR2, FGFR3, FGFR4, etc. (<NPL>.

RET is also a member of RTK family and its normal physiological functions include renal development, development of the nervous system, maintenance and renewal of sperm stem cells, differentiation of myelomonocytic cells, formation of lymphoid tissues, etc.. RET is expressed in human intestinal ganglion cells, neuroblastoma, pheochromocytoma, medullary thyroid carcinoma, thyroid C cells, and melanocytes, etc. In recent years, based on intensive study on RET, it has been found that overactivation of RET in tumors significantly promotes proliferation, survival, invasion, metastasis, and tumor inflammation of various tumors (<NPL>). For example, RET point mutation is up to <NUM>% in patients with medullary thyroid carcinoma; RET gene rearrangement accounts for <NUM>% to <NUM>% in patients with papillary thyroid cancer ; and RET is also overexpressed in adenocarcinoma, colon cancer, pancreatic cancer, breast cancer, acute leukemia.

Currently, the marketed drugs as a multi-targeted inhibitor having FGFR and RET inhibitory activity mainly target vascular endothelial growth factor receptor <NUM> (VEGFR2, also known as KDR), such as Regorafenib. Studies have shown that the strong inhibition of KDR causes cancer patients to have strong cardiovascular side effects, such as thrombotic microangiopathy, hypertension, congestive heart failure, coagulopathy, pancreatitis and so on. Based on current studies, there is few report on RET inhibitors with strong selectivity. Meanwhile, the inevitable drug resistance problem in other kinase inhibitors also exists in RET inhibitors. For example, the classic gatekeeper site mutations - RET V804M and V804L have been discovered. Currently, preclinical studies have shown that few inhibitors have the potential to overcome resistance.

The compounds disclosed in the Boral Sougato's patent must have sulfone imine (<CIT>, <CIT>, <CIT>, <CIT>) or tetrazolium (<CIT>) as advantageous structure at meta-position of the pyridine ring, and focus on VEGFR. The compounds have low druggability, low exposure in vivo, and fail to achieve anti-tumor effect in vivo.

Kassoum Nacro's patent discloses a series of amino-substituted nitrogen-containing heteroaromatic ring (<CIT>) whose target is tyrosine kinase MNK. However, as for A ring, it doesn't specifically disclose o-amino substituted heterocycle.

<CIT> disclosed a class of alkynyl heterocyclic compounds and uses thereof. However, o-amino substituted heteroaryl ring is not disclosed either.

The present invention provides a novel o-aminoheteroaryl alkynyl-containing compound, preparation method therefor and use thereof.

The present invention is implemented by the following technical solutions:
A compound of formula (I), or a deuterated compound, or a pharmaceutically acceptable salt thereof:
<CHM>
as defined in claim <NUM>.

Preferably, in the above compound of formula (I) or the deuterated compound, or pharmaceutically acceptable salt thereof, the preferred compound, or the deuterated compound, or pharmaceutically acceptable salt thereof is selected from the following compounds:
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>
<CHM>.

The present invention also provides a method for preparing the compound of formula (I), or the deuterated compound, or pharmaceutically acceptable salt thereof, which comprises a step of reacting a compound of formula (<NUM>) with a compound of formula (<NUM>)
<CHM>
wherein, each of R and R<NUM>-R<NUM> is independently defined as above.

Preferably, it comprises: in the presence of a transition metal palladium and copper catalyst and in alkaline condition, coupling the compound of formula (<NUM>) with the compound of formula (<NUM>). Preferably, the palladium catalyst comprises Pd(PPh<NUM>)<NUM>Cl<NUM>, Pd(OAc)<NUM>, and/or Pd(PPh<NUM>)<NUM> Preferably, the copper catalyst comprises CuI and/or CuCl. Preferably, the base used for the alkaline condition comprises one or more bases selected from CsF, Cs<NUM>CO<NUM>, K<NUM>CO<NUM>, triethylamine, diisopropylethylamine, and DMAP. Preferably, the solvent for coupling reaction comprises one or more solvents selected from acetonitrile, <NUM>,<NUM>-dioxane, and DMF.

More preferably, the method comprises a step of reacting the compound of formula (<NUM>) with the compound of formula (<NUM>) in the presence of cesium fluoride, Pd(PPh<NUM>)<NUM>Cl<NUM>, CuI and triethylamine and in acetonitrile as a solvent.

More preferably, the method comprises any of the following Schemes I or II:
<CHM>.

The present invention also provides a pharmaceutical composition comprising one or more of the above compounds of formula (I) or the deuterated compound, or the pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

The present invention also provides the use of the above formula (I) compound or the deuterated compound, or the pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in preparation of FGFR kinase inhibitor, RET kinase inhibitor and/or inhibitor for mutant of FGFR or RET kinases.

The present invention also provides the use of the above compound of the formula (I) or the deuterated compound, or pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in preparing a medicament for treating tumor; optionally, the tumor comprises non-small cell lung cancer, breast cancer, thyroid cancer (medullary thyroid carcinoma, papillary thyroid cancer), gastric cancer, bladder cancer, endometrial cancer, prostate cancer, cervical cancer, colon cancer, esophageal cancer, keratinoma, myeloma, rhabdomyosarcoma, acute leukemia, liver cancer, adenocarcinoma, and pancreatic cancer.

The present invention also provides the use of the above compound of formula (I) or the deuterated compound thereof, or pharmaceutically acceptable salt thereof, or the above pharmaceutical composition in treating tumor. Optionally, the tumor comprises non-small cell lung cancer, breast cancer, thyroid cancer (medullary thyroid carcinoma, papillary thyroid cancer), gastric cancer, bladder cancer, endometrial cancer, prostate cancer, cervical cancer, colon cancer, esophageal cancer, keratinoma, myeloma, rhabdomyosarcoma, acute leukemia, liver cancer, adenocarcinoma, and pancreatic cancer.

According to an embodiment of the present invention, the o-aminoheteroarylalkynyl-containing compound has an advantage of high dual-targeting inhibitory activity on FGFR and RET.

According to another embodiment of the present invention, the o-aminoheteroarylalkynyl-containing compound has an advantage of low KDR activity.

According to another embodiment of the present invention, the o-aminoheteroarylalkynyl-containing compound exhibits strong inhibition of cell proliferation activity in human lung cancer NCI-H1581, gastric cancer cell line SNU16 and RET-dependent sensitive cell line BaF3-CCDC6-Ret and mutants thereof.

According to another embodiment of the present invention, the pharmacokinetic data indicate that the o-aminoheteroarylalkynyl-containing compound has good druggability and exhibits significant inhibitory activity on tumor growth in a long-term animal pharmacodynamic model.

According to another embodiment of the present invention, the animal is in good condition (including no significant decrease in body weight) at the effective dose, and no significant toxicity of other RTK multi-target inhibitors is observed (no animal death and molting).

Specific embodiments of the present invention will be described in detail below. It should be understood that the specific embodiments described herein are illustrative of the invention and are not intended to limit the invention.

<NUM>-methyl-<NUM>-nitrobenzotrifluoride (<NUM>, <NUM> mmol), NBS(<NUM> ,<NUM> mmol) , AIBN(<NUM>, <NUM> mmol ) and CCl<NUM> (<NUM>) were added into a round-bottom flask and the reaction was carried out by heating in an oil bath under <NUM> for <NUM> hours until completion. The mixture was cooled to room temperature, the solvent was removed under reduced pressure. After column chromatography, the product <NUM>-trifluoromethyl-<NUM>-nitrobenzyl bromide was obtained (<NUM>, yield: <NUM>%).

<NUM>-trifluoromethyl-<NUM>-nitrobenzyl bromide (<NUM> , <NUM> mmol), N-methylpiperazine (<NUM> , <NUM> mmol ), Et<NUM>N (<NUM>, <NUM> mmol ) and CH<NUM>Cl<NUM> (<NUM>) were added into a round-bottom flask, and the reaction was carried out at room temperature for <NUM> hours until completion. The solvent was removed under vacuo. After column chromatography, the product <NUM>-methyl-<NUM>-(<NUM>-nitro-<NUM>-(trifluoromethyl)benzyl)piperazine is obtained (<NUM>, yield: <NUM>%).

<NUM>-methyl-<NUM>-(<NUM>-nitro-<NUM>-(trifluoromethyl)benzyl)piperazine (<NUM> , about <NUM> mmol), reducing Fe powder (<NUM>, <NUM> mmol), NH<NUM>Cl (<NUM>, <NUM> mmol), EtOH(<NUM>) were added into a round bottom flask; and the reaction was carried out at <NUM> in an oil bath for <NUM> hours until completion. After filtration via a pad of celite, the filtrate was concentrated under reduced pressure. The product <NUM>-((<NUM>-methyl piperazin-<NUM>-yl)methylene)-<NUM>-(trifluoromethyl)aniline (<NUM> , yield: <NUM>%) was obtained by column chromatography.

<NUM>-iodo-<NUM>-chlorobenzoic acid (<NUM>, <NUM> mmol ), Et<NUM>N (<NUM>, <NUM> mmol), HATU (<NUM> , <NUM> mmol), DMF (<NUM> ) were added into a round-bottom flask. After stirring at room temperature for <NUM> hour, <NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoromethyl)aniline (<NUM>, <NUM>) was added. The reaction was carried out at room temperature for <NUM> hours until completion, and the solvent was removed under reduced pressure. <NUM>-iodo-<NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoromethyl)phenyl)benzami de (<NUM>, yield: <NUM>%) was obtained by column chromatography.

<NUM>-iodo-<NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoromethyl)phenyl)ben zamide (<NUM>, <NUM> mmol), trimethylsilylacetylene (<NUM>, <NUM> mmol), Pd (PPh<NUM>)<NUM>Cl<NUM> (<NUM>, <NUM> mmol), CuI (<NUM>, <NUM> mmol), Et<NUM>N (<NUM>, <NUM> mmol)) and MeCN (<NUM>) were added into a round-bottom flask, and the mixture was heated to <NUM> in oil bath and reacted overnight until the reaction was completed. After column chromatography, the product <NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoromethyl)phenyl)-<NUM>-((trimethylsil yl)ethynyl)benzamide was obtained (<NUM>, yield: <NUM>%).

<NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoromethyl)phenyl)-<NUM>-((Trimet hylsilyl)ethynyl)benzamide (<NUM>, <NUM> mmol), <NUM>-amino-<NUM>-iodopyridine (<NUM>, <NUM> mmol), Pd (PPh<NUM>)<NUM>Cl<NUM> (<NUM>, <NUM> mmol), CuI (<NUM>, <NUM> mmol), CsF (<NUM>, <NUM> mmol), Et<NUM>N (<NUM>, <NUM> mmol) and MeCN (<NUM>) were added into a round-bottom flask, and mixture was heated to <NUM> in oil bath and reacted overnight until the reaction was completed. After column chromatography, the product <NUM>-(<NUM>-aminopyridine-<NUM>-ethynyl)-<NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoro methyl)phenyl)benzamide (huFGFR267) was obtained (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM> ), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-iodo-<NUM>-fluorobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-iodo-<NUM>-methylbenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M +<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-iodo-<NUM>-methoxybenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (td , J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-iodo-<NUM>-cyanobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-iodobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-fluoro-<NUM>-iodobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM> , <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>) , <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-iodo-<NUM>-chlorobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, "J =<NUM>, <NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>) ), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-iodo-<NUM>-trifluoromethylbenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> ( Dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>) , <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m , <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, "J =<NUM>, <NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>) , <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-methyl-<NUM>-iodobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>) ). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-methoxy-<NUM>-iodobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-fluoropyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (ddt, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (m, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-chloropyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-methylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-cyclopropylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-trifluoromethylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-cyanopyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

NMR <NUM> (d, J=<NUM>, <NUM>), , <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-methoxypyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

NMR <NUM> (s, <NUM>), <NUM> (s, <NUM>) (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-trifluoromethoxypyridine was used in place of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM> , <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-cyclopropyloxypyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM> , <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m , <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - -<NUM> (m, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-(<NUM>-oxetanyl)oxypyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-(<NUM>-hydroxyethyl)oxypyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>) , <NUM> (td, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-(<NUM>-methoxyethyl)oxypyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

NMR <NUM> (d, J=<NUM>, <NUM>), <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (td, J=<NUM>, <NUM>, <NUM>), <NUM> (td, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-(<NUM>-methylaminoethyl)oxypyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m , <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-(<NUM>-dimethylaminoethyl)oxypyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+ <NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-hydroxymethylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>) , <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM> ).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-methoxymethylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s , <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-trifluoromethoxymethylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-methylaminomethylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s , <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-dimethylaminomethylpyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-methylaminopyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) ) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-dimethylaminopyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>) , <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-acetylaminopyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>) , <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>)), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-(<NUM>-cyclopropylacetyl)aminopyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>). LR -MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-acrylamidopyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM> NMR (<NUM>, CDCl3 ) δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>) ), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-(<NUM>-dimethylamino-<NUM>-alkenylbutanoyl)aminopyridine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM> Hz, <NUM>), <NUM> (d, J=<NUM> Hz, <NUM>) , <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM> Hz, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

Compound <NUM>-methyl-<NUM>-piperidinol (<NUM>, <NUM> mmol) and NaH (<NUM>, <NUM> mmol), were added into a round-bottom flask using DMF as a solvent. The mixture was stirred in ice water bath for <NUM>, and <NUM>-fluoro-<NUM>-nitrotrifluorotoluene (<NUM>, <NUM> mmol) was added, and the reaction was carried out at room temperature for <NUM> hours. The product <NUM>-methyl-<NUM>-(<NUM>-nitro-<NUM>-(trifluoromethyl)phenylhydroxy)piperidine (<NUM>, yield: <NUM>%) was obtained by purification.

Compound <NUM>-methyl-<NUM>-(<NUM>-nitro-<NUM>-(trifluoromethyl)phenylhydroxy)piperidine (<NUM> , <NUM> mmol), Fe powder (<NUM>, <NUM> mmol), AcOH (<NUM>, <NUM> mmol), and ethanol (solvent) were added into a round-bottom flask, the reaction was carried out at <NUM> for <NUM> hours until completion, and the product <NUM>-((<NUM>-methylpiperidinyl-<NUM>-yl)hydroxy)-<NUM>-(trifluoromethyl)aniline (<NUM>. , Yield: <NUM>%) was obtained by purification.

<NUM>-iodo-<NUM>-fluorobenzoic acid (<NUM>, <NUM> mmol), Et<NUM>N (<NUM>, <NUM> mmol), and HATU (<NUM>, <NUM> mmol) were added into a round bottom flask, and DMF ( <NUM>) was added successively. After stirring at room temperature for <NUM> hr, <NUM>-((<NUM>-methylpiperidinyl-<NUM>-yl)hydroxy)-<NUM>-(trifluoromethyl)aniline (<NUM>, <NUM>) was added, and the reaction was carried out at room temperature for <NUM> hours until completion. The solvent was evaporated to dryness under reduced pressure. After column chromatography, <NUM>-chloro-<NUM>-iodo-N-(<NUM>-((<NUM>-methylpiperidinyl-<NUM>-yl)hydroxy)-<NUM>-(trifluoromethyl)phenyl)benzamid e (<NUM>, yield: <NUM>%) was obtained.

<NUM>-chloro-<NUM>-iodo-N-(<NUM>-((<NUM>-methylpiperidin-<NUM>-yl)hydroxy)-<NUM>-(trifluoromethyl)phenyl)benza mide (<NUM>, <NUM> mmol)), trimethylsilylacetylene (<NUM>, <NUM> mmol), Pd (PPh<NUM>) <NUM> Cl<NUM>(<NUM>, <NUM> mmol), CuI(<NUM>, <NUM> mmol), Et<NUM>N (<NUM>, <NUM> mmol)) and MeCN (<NUM>) were added into a round-bottom flask, and the reaction was carried out overnight at <NUM> in oil bath until completion. After column chromatography, the product <NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperidin-<NUM>-yl)hydroxy)-<NUM>-(trifluoromethyl)phenyl-<NUM>-((trimethylsilyl)e thynyl)benzamide was obtained (<NUM>, yield: <NUM>%).

<NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoromethyl)phenyl)-<NUM>-((trimeth ylsilyl)ethynyl)benzamide (<NUM>, <NUM> mmol), <NUM>-amino-<NUM>-iodopyridine (<NUM>, <NUM> mmol), Pd (PPh<NUM>)<NUM>Cl<NUM> (<NUM>, <NUM> mmol), CuI (<NUM>, <NUM> mmol), CsF (<NUM>, <NUM> mmol), Et<NUM>N (<NUM>, <NUM> mmol) and MeCN (<NUM>)were added into a round-bottom flask, and the reaction was carried out overnight at <NUM> in oil bath until completion. After column chromatography, the product <NUM>-(<NUM>-aminopyridine-<NUM>-ethynyl)-<NUM>-chloro-N-(<NUM>-((<NUM>-methylpiperazin-<NUM>-yl)methylene)-<NUM>-(trifluoro methyl)phenyl)-<NUM>-((trimethylsilyl)ethynyl)benzamide was obtained (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (ddd, J=<NUM>, <NUM>, <NUM>, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> ( Dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, "J =<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, "J =<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (p, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-iodo-<NUM>-methoxybenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>). LR -MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodopyrazine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CD<NUM>OD) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>) , <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-fluoropyrazine was used instead of <NUM>-amino-<NUM>-iodopyridine and <NUM>-iodobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J =<NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (dt, J=<NUM>, <NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>)), <NUM> (d, J"=<NUM>, <NUM>), <NUM> (s, <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-hydroxymethylpyrazine was used instead of <NUM>-amino-<NUM>-iodopyridine and <NUM>-iodobenzoic acid was used instead of <NUM>-iodo-<NUM>-chlorobenzoic acid.

NMR <NUM> (s, <NUM>), <NUM> (s, <NUM>) (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>). LR- MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis method was carried out as Example <NUM>, except that <NUM>-amino-<NUM>-iodo-<NUM>-fluoropyrazine was used instead of <NUM>-amino-<NUM>-iodopyridine.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (ddd, J =<NUM>, <NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>) , δ-MS (ESI) m/z <NUM> (M+<NUM>).

The synthesis was carried out as Example <NUM>, except that <NUM>-tert-butoxycarbonylpiperazine was used instead of N-methylpiperazine.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>). (ESI) m/z <NUM> (M+<NUM>).

HuFGFR459 (<NUM>, <NUM> mmol) was dissolved in anhydrous dichloromethane (<NUM>), and trifluoroacetic acid (<NUM>) was added dropwise into the solution under ice bath condition. The reaction was carried out in ice bath for <NUM>. After purification, the product HuFGFR472 was obtained (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>). (ESI) m/z <NUM> (M+<NUM>).

HuFGFR267 (<NUM> , <NUM> mmol) was dissolved in anhydrous methanol, and <NUM> hydrogen chloride in methanol (<NUM>) was added dropwise into the solution under ice bath condition. The reaction was carried out for <NUM> at room temperature, and the solvent was evaporated to obtain HuFGFR459 (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m , <NUM>). LR-MS (ESI) m/z <NUM> (M+<NUM>).

HuFGFR472 (<NUM> , <NUM> mmol) was dissolved in anhydrous DMF, potassium carbonate (<NUM>, <NUM> mmol) was successively added, and then deuterated iodomethane (<NUM>, <NUM> mmol) was added under ice bath condition. The reaction was carried out for <NUM> in ice bath to obtain HuFGFR474 (<NUM>. g, Yield: <NUM>%).

<NUM>H NMR (<NUM>, DMSO) δ <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> (dd, J=<NUM>, <NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>). LR-MS (ESI) m/z <NUM> (M +<NUM>).

The structures of compound A34, HuFGFR143, HuFGFR148, HuFGFR150, HuFGFR151, Ponatinib (or AP24534) and LY2874455 are as follows:
<CHM>.

Enzyme reaction substrate Poly(Glu, Tyr)<NUM>:<NUM> was diluted with potassium-free PBS (<NUM> sodium phosphate buffer, <NUM> NaCl, pH <NUM>-<NUM>) to <NUM>µg/mL. The enzyme label plate was coated with <NUM>µL/well. The reaction was conducted under <NUM> for <NUM>-<NUM> hours. After the liquid was removed from the wells, the plate was washed three times with <NUM>µL / well of T-PBS (PBS containing <NUM>% Tween-<NUM>), each for <NUM> minutes. The enzyme plate was dried in <NUM> dryer for <NUM>-<NUM> hours.

50µL ATP solution diluted with buffer (<NUM> HEPES pH <NUM>, <NUM> MgCl<NUM>, <NUM> MnCl<NUM>, <NUM> Na<NUM>VO<NUM>, <NUM> DTT) to a final concentration of <NUM> was added into each well. The compound was diluted with DMSO to a suitable concentration (<NUM>µL/well) or the well contained the corresponding concentration of DMSO (negative control well) Then various kinase recombinant proteins diluted with <NUM>µL of reaction buffer was added to initiate the reaction. Each experiment required duplicate enzyme-free control well. The reaction was carried out for <NUM> hour on a <NUM> Shaker (<NUM> rpm). The plate was washed three times with T-PBS. A dilution of the primary antibody PY99 was added (<NUM>µL/well), and the reaction was conducted in a shaker at <NUM> for <NUM> hr. The plate was washed three times with T-PBS. A dilution of the horseradish peroxidase-labeled goat anti-mouse IgG secondary antibody was added (<NUM>µL/well), and the reaction was conducted in a shaker at <NUM> for <NUM> hour. Plate was washed with T-PBS for three times. <NUM>/mL OPD coloration solution (diluted with <NUM> citric acid-sodium citrate buffer containing <NUM>% H<NUM>O<NUM> (pH=<NUM>)) was added (<NUM>µL / well), reacted for <NUM>-<NUM> minutes at <NUM> in dark. The reaction was quenched with <NUM> H<NUM>SO<NUM> (<NUM>µL/well), and read at <NUM> using a tunable microplate reader SPECTRA MAX <NUM>.

The inhibition ratio of the sample was determined by the following formula:<MAT>.

The IC<NUM> values were obtained via four-parameter regression analysis using the software supplied with the microplate reader.

The enzyme activity data of the compounds prepared in the present invention, compound HuFGFR151, compound HuFGFR117, the positive control Ponatinib and the positive control LY2874455 against the three enzymes of FGFR1, RET and KDR are listed in Table <NUM>:.

Experimental results: As seen from Table <NUM>, in the evaluation of biological activity, the o-aminoheteroaryl alkynyl-containing compounds of the present invention have a high activity of inhibiting FGFR1 and RET kinase at a concentration of <NUM>, while the compounds prepared in the examples of the present invention have low KDR activity. The activity of these compounds on KDR is significantly weaker than that on FGFR1 or RET, thus indicating these compounds have clear selectivity which is beneficial to solve the technical problems of hepatic toxicity and cardiotoxicity of Panatinib. Compared with the compounds of the present invention, compound HuFGFR151 has a different amino position, which results in the significant decrease of its inhibitory activity on FGFR1 and RET kinases. Compared with compound A34, HuFGFR143, HuFGFR148, and HuFGFR150, the introduction of o-amino in the compounds of the present invention results a significant increase in the activity against FGFR1 and RET kinases and selectivity. However, the KDR activity of the positive control drugs (Ponatinib and LY2874455) is high.

The inhibitory activity data of the compounds HuFGFR267 and HuFGFR293 against the RET-related mutant enzyme are shown in Table <NUM>, wherein, Ret (V804M) is a commercially available recombinant protein. The results showed that HuFGFR267 and HuFGFR293 had significant inhibitory activities against Ret and Ret (V804M), especially for Ret and its V804M mutant kinase.

The cells were seeded into a <NUM>-well plate (<NUM>,<NUM>/well). After incubation for <NUM>-<NUM> hours, the compounds were added to react for <NUM> hours, and then the cells were collected and firstly washed once with cold PBS (containing <NUM> mmol sodium vanadate); then <NUM>×SDS gel loading buffer (<NUM> mmol Tris-HCl (pH <NUM>), <NUM> mmol DTT, <NUM>% SDS, <NUM>% glycerol, <NUM> mmol sodium vanadate , <NUM>% bromophenol blue) was added to lyse cells. The cell lysate was heated in a boiling water bath for <NUM> minutes, and then centrifuged at <NUM>,<NUM> rpm for <NUM> minultes at <NUM>.

The supernatant was taken for SDS-PAGE electrophoresis (Mini-PROTEAN <NUM> Cell, Bio-Rad, Hercules, CA, US A). After electrophoresis, the proteins were transferred to a nitrocellulose membrane using a semi-dry electrotransfer system (Amersham Life Sciences, Arlington Heights, IL, USA). The nitrocellulose membrane was placed in a blocking solution (<NUM>% skim milk powder diluted in TBS containing <NUM> mmol sodium vanadate) for <NUM> hours at room temperature, and then the membrane was placed and reacted with a primary antibody at <NUM> ° C overnight. The membrane was washed three times with TBS containing <NUM> mmol sodium vanadate for <NUM> each time. The membrane was placed in a secondary antibody solution and reacted for <NUM>-<NUM> hours at room temperature. After the membrane was washed three times as above, the membrane was stained using ECL (Picece, Rockford, IL) reagent was used for staining and then developed.

The results that Compound HuFGFR267 (<NUM>), HuFGFR293 (<NUM>) and positive control Ponatinib inhibited RET phosphorylation and the downstream signaling pathway in tumor cells and tool cell lines were shown in <FIG>. As seen from <FIG>, the o-aminoheteroaryl alkynyl-containing compound of the present invention targeted to and significantly inhibited the activation of the RET signaling pathway at the cellular level.

The AZD4547 structure is as follows:
<CHM>.

The tumor tissue in the vigorous growth period was cut into about <NUM><NUM>, and inoculated subcutaneously in the right axilla of nude mice under aseptic conditions. The diameter of the subcutaneously implanted tumor in nude mice was measured by a vernier caliper. The animals were randomly divided into groups when the average volume was about <NUM><NUM>. In the compound HuFGFR267 <NUM>/kg group, the compound was formulated with <NUM>% methylcellulose (MC) to the required concentration before use. The formulation of compound was prepared once a week, and orally administered once a day for <NUM> days. The positive control drug AZD4547 was diluted to the required concentration with water for injection containing <NUM>% Tween <NUM> before use. The formulation of AZD4547 was prepared once a week, and orally administered once a day for <NUM> days. In the solvent control group, an equal amount of water for injection was administrated. The diameter of the transplanted tumor was measured twice a week during the entire experiment, and the body weight of the mice was weighed. The tumor volume (TV) was calculated as: TV=<NUM>/<NUM> × a × b<NUM>, where a and b represented length and width, respectively. The relative tumor volume (RTV) was calculated based on the measured results, and the formula was: RTV=V t / V<NUM>, wherein V<NUM> represented the tumor volume obtained when the mice was divided and administered (i.e., d<NUM>), and Vt represented the tumor volume at each measurement. The evaluation index of antitumor activity was: relative tumor proliferation rate T/C (%), and the formula was as follows: T/C (%)=(T RTV / C RTV) × <NUM>%, wherein T RTV was TRV in treatment group; and C RTV was RTV in negative control group.

The results of the inhibitory effect of the compound HuFGFR267 on growth of xenografts of human lung cancer NCI-H1581 in nude mice are shown in Table <NUM> and <FIG>, wherein the data in Table <NUM> correspond to numerical points in the curve of <FIG>. As seen from <FIG>, in HuFGFR267 <NUM>/kg group, after orally administered once a day for <NUM> days, the growth of subcutaneously xenografts of human lung cancer NCI-H1581 in nude mice was significantly inhibited, and T/C obtained on the 14th day was <NUM>%. In the positive control AZD4547 <NUM>/kg group, it was administrated in the same way as above, and the growth of subcutaneously xenografts of human lung cancer NCI-H1581 in nude mice was significantly inhibited and the T/C obtained on the 14th day was <NUM>%. During the experiment, no mice died, and the mice in each group were in good condition. It can be seen from <FIG> and Table <NUM> (wherein the data in Table <NUM> correspond to numerical points in the curve of <FIG>), the body weight of mice bearing human lung cancer NCI-H1581 tumor in the compound HuFGFR267 group had no significant change. Thus, it indicated that the o-aminoheteroaryl alkynyl-containing compound of the present invention had a significant inhibitory effect on the growth of subcutaneous xenografts of human lung cancer NCI-H1581 in nude mice, and advantage of low toxicity.

The results of the inhibitory effect of compound HuFGFR267 on growth of human gastric cancer SNU-<NUM> xenografts in nude mice are shown in <FIG>, wherein the data of Table <NUM> correspond to numerical points in the curve of <FIG>. In HuFGFR267 <NUM>/kg and <NUM>/kg groups, compounds were administered orally once a day for <NUM> days, and the growth of human gastric cancer SNU-<NUM> xenografts in nude mice was which significantly inhibited. The T/C values obtained on day <NUM> were <NUM>% and <NUM>%, respectively. In the positive control AZD4547 <NUM>/kg group, AZD4547 was administrated in the same way as above and the growth of subcutaneous xenografts of human gastric cancer SNU-<NUM> in nude mice was significantly inhibited. T/C obtained on day <NUM> was <NUM>%. During the experiment, no mice died, and the mice in each group were in good condition. As seen from <FIG> (the data in Table <NUM> correspond to each numerical points in the curve of <FIG>), body weight of mice bearing human gastric cancer SNU-<NUM> tumor in the compound HuFGFR267 group had no significant change. Thus, it indicated that the o-aminoheteroaryl alkynyl-containing compound of the present invention had a significant inhibitory effect on the growth of subcutaneous xenografts of human gastric cancer SNU-<NUM> in nude mice, and advantage of low toxicity.

The tumor tissue in the vigorous growth period was cut into <NUM><NUM>, and inoculated subcutaneously in the right axilla of nude mice under aseptic conditions. The diameter of the xenograft in nude mice was measured with a vernier caliper. The animals were randomly divided into groups when the average tumor volume was grown to about <NUM> <NUM>. In HuFGFR1-<NUM> and HuFGFR1-<NUM> groups ( <NUM>/kg and <NUM>/kg) the compounds were orally administered once a day for <NUM> consecutive days. In the positive control drug Ponatinib <NUM>/kg group, Ponatinib was orally administered once a day for <NUM> days. In the solvent control group, mice were gave an equal amount of solvent. The diameter of the xenograft was measured twice a week during the entire experiment, and the body weight of the mice was weighed. The tumor volume (TV) was calculated as: TV=<NUM>/<NUM> × a × b<NUM>, where a and b represented length and width, respectively. The relative tumor volume (RTV) was calculated based on the measured results, and the formula was: RTV=Vt / V<NUM>, wherein V<NUM> represented the tumor volume obtained when the mice was divided and administered (i.e., d<NUM>), and Vt represented the tumor volume at each measurement. The anti-tumor activity evaluation index was the relative tumor proliferation rate T/C (%), and the calculation formula was as follows: T/C (%)=(T RTV / C RTV) × <NUM>%, wherein T RTV was TRV in treatment group; and C RTV was RTV in negative control group.

The experimental results are shown in <FIG> (where the data in Tables <NUM> and <NUM> correspond to the numerical values in the curves of <FIG>, respectively). HuFGFR1-<NUM><NUM>/kg group showed significant toxicity, and administration was stopped due to the mice were found to have a poor state with low temperature on the second day. One mouse died on the third day. Two mice died on the fifth day. The compound was administrated again because the mouse was recovered on day <NUM>. In HuFGFR1-<NUM><NUM>/kg group, the compound was administered orally once a day for <NUM> days, and the growth of human gastric cancer SNU-<NUM> in nude mice was significantly inhibited. The T/C obtained on day <NUM> was <NUM>%, but one mouse died on day <NUM>. In the compound HuFGFR1-<NUM><NUM>/kg group, the growth of subcutaneous xenograft of human gastric cancer SNU-<NUM> in nude mice was significantly inhibited. The T/C obtained on day <NUM> was <NUM>%, but one mouse died on the 9th and 10th day respectively, and other mice showed dry skin, molting, and poor condition, so the administration was stopped. The administration was started again because the mice was recovered and the molting disappeared and the skin returned to normal on day <NUM>. In HuFGFR1-<NUM><NUM>/kg group, it was orally administered once a day, and the growth of subcutaneous xenografts of human gastric cancer SNU-<NUM> in nude mice was weakly inhibited. The T/C obtained on day <NUM> was <NUM>%, and the tumor inhibition rate was low. In the Ponatinib <NUM>/kg group, it was orally administered once a day for <NUM> days, and the growth of subcutaneous xenografts of human gastric cancer SNU-<NUM> in nude mice was significantly inhibited. The T/C obtained on day <NUM> was <NUM>%.

Experimental conclusion: The HuFGFR267 of the present invention significantly inhibited the growth of subcutaneous xenografts of human gastric cancer SNU-<NUM> in nude mice, and the T/C obtained on day <NUM> was <NUM>% and <NUM>%, respectively. The weight of the tumor-bearing mice did not change significantly and the mice were in good condition. Compound HuFGFR1-<NUM> (morpholine substituted in meta-position of pyridine) which was structurally similar to compound HuFGFR267 of the present invention, had a weaker inhibition on growth of subcutaneous xenografts of human gastric cancer SNU-<NUM> in nude mice at the dosage of <NUM>/kg than that of HuFGFR267, and the T/C obtained on day <NUM> was <NUM>%, while the weight of the tumor-bearing mice decreased significantly. One mouse died on day <NUM>. The dosage of <NUM>/kg showed great toxicity. The mice were found to have poor state with low temperature on the second day. One mouse died on the third day. Two mice died on the fifth day. Another compound HuFGFR1-<NUM> (without amino substitution in the o-position of pyridine) which was structurally similar to HuFGFR267 of the present invention, has a weaker inhibitory effect on the growth of subcutaneous xenografts of human gastric cancer SNU-<NUM> in nude mice than that of HuFGFR267, and T/C obtained on day <NUM> were <NUM>% and <NUM>%, while the weight of the tumor-bearing mice was significantly decreased. One mouse died on the 9th and 10th day respectively, and other mice appeared dry skin, molting and poor state. It indicates that the introduction of the o-amino of the pyridine in the present invention can effectively increase the tumor inhibitory activity of compounds and exhibit a significant advantage of low toxicity.

Ponatinib (AP24534) was administered intravenously (IV) and orally (PO) to SD rats. Blood samples were taken at different time points. LC-MS/MS was used to determine the concentration of compound in the plasma of rats after administration of the test compound, and the relevant pharmacokinetic parameters were calculated to examine oral bioavailability and pharmacokinetic properties of the compound in rats. The results are shown in Table <NUM>.

Compound HuFGFR267 was administered intravenously and orally to SD rats. Blood samples were taken at different time points. LC-MS/MS was used to determine the concentration of compound in the plasma of rats after administration of the test compound, and the relevant pharmacokinetic parameters were calculated to examine oral bioavailability and pharmacokinetic properties of the compound in rats. The results are shown in Table <NUM>.

It can be seen from Table <NUM> and Table <NUM> that compound HuFGFR267 is superior to the marketed drug AP24534 in the aspect of exposure, thus having a better potential for developing into a medicine.

In summary, the o-aminoheteroaryl alkynyl-containing compounds in the examples of the present invention have advantages of a high FGFR and RET dual-targeting inhibitory activity and a relatively low KDR activity. The compound of formula (I) exhibits a strong inhibitory activity on human lung cancer cell line NCI-H1581 and gastric cancer cell line SNU16 as well as an RET-dependent sensitive cell line BaF3-CCDC6-Ret and mutants thereof. Pharmacokinetic data has shown that the o-aminoheteroaryl alkynyl-containing compound has good druggability, and exhibits significant inhibition on the growth of related tumors in a long-term animal model while in the efficacy dosage, the animal has a good condition (including no significant decrease in body weight), and no significant toxicity is observed (no animal death or molting).

Claim 1:
A compound of formula (I), or a deuterated compound, or a pharmaceutically acceptable salt thereof:
<CHM>
wherein:
R is amino;
M is C or N, and when M is N, R<NUM> is none;
R<NUM> is selected from the group consisting of -H, halogen, hydroxyl, cyano, C<NUM>-<NUM> alkyl (optionally substituted by halogen, hydroxyl, C<NUM>-<NUM> alkoxy, trifluoromethoxyl, mono or di C<NUM>-<NUM> alkylamino), C<NUM>-<NUM> alkoxy (optionally substituted by halogen, hydroxyl, C<NUM>-<NUM> alkoxyl, amino, mono or di C<NUM>-<NUM> alkylamino), amino, mono or di C<NUM>-<NUM> alkylamino, C<NUM>-<NUM> alkylamido, C<NUM>-<NUM> cycloalkylamido, and C<NUM>-<NUM>alkenylamido optionally substituted by mono or di C<NUM>-<NUM> alkylamino;
R<NUM> is selected from -H, or halogen;
R<NUM> is selected from -H, halogen, cyano, an optionally halogenated C<NUM>-<NUM> alkyl, or C<NUM>-<NUM> alkoxy;
R<NUM> is a C<NUM>-<NUM> alkyl or oxyl substituted by a <NUM>- or <NUM>-membered aliphatic heterocyclyl having <NUM>-<NUM> N atoms on the ring, wherein the <NUM>- or <NUM>-membered aliphatic heterocyclyl is optionally substituted by C<NUM>-<NUM> alkyl;
each of theheteroaryl, heteroaryl ring is independently and optionally substituted by one or more substituents selected from the group consisting of halogen, oxo, and alkyl;
the alkyl is a saturated aliphatic straight or branched alkyl group having <NUM>-<NUM> carbon atoms;
the halogen is each independently selected from the group consisting of F, Cl, Br, and I;the heteroaryl or heteroaryl ring is a <NUM>-<NUM> membered aromatic monocyclic or fused bicyclic ring having one or more heteroatoms selected from N, O, and S;
the cycloalkyl is a saturated or unsaturated <NUM>-<NUM> membered monocyclic or polycyclic alicyclic ring;
the cycloalkylene is a saturated or unsaturated <NUM>-<NUM> membered monocyclic or polycyclic aliphatic cycloalkylene;
the heterocyclyl is a saturated or unsaturated <NUM>-<NUM> membered monocyclic or polycyclic aliphatic heterocycle containing one or more heteroatoms selected from N, O, and S;
the heterocyclylene is a saturated or unsaturated <NUM>-<NUM> membered monocyclic or polycyclic aliphatic heterocyclylene containing one or more heteroatoms selected from N, O, and S;
preferably, the pharmaceutically acceptable salt comprises hydrochloride, methanesulfonate, maleate or the like.