Spiro compound and application thereof

Abstract

The invention provides a spiro compound which is shown in a formula I and has a binding effect with E3 ligase protein CRBN, and application of the spiro compound in preparation of a medicine for treating cell abnormal proliferation diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to a novel spirocyclic ligand compound that binds to the cereblon E3 ubiquitin ligase protein. Background Art

[0002] Protein degradation is a highly regulated process essential for maintaining cellular homeostasis. The selective identification and removal of damaged, misfolded, or excessive proteins are achieved through the ubiquitin-proteasome pathway (UPP). The UPP is responsible for clearing defective proteins and is characterized by being ATP-dependent, highly efficient, and highly selective. Its catalytic part is the ubiquitination E3 ligase, but it needs to be first recruited to the protein to be degraded. The PROTACs technology is designed based on the UPP principle, connecting the ligand of the target protein and the ligand of the E3 ligase with a suitable chemical bond, so that it can recognize the target protein and enhance the binding ability of the ligase E3 to the target protein, and then target ubiquitination to force the degradation of the target protein, and has the characteristics of catalytic dosage, high efficiency, and high selectivity.

[0003] Multiple ubiquitin molecules covalently link to the terminal lysine residues through the E3 ubiquitin ligase to label the protein for proteasomal degradation, where the protein is digested into small peptides and ultimately into its constituent amino acids, and the amino acids are used as building blocks for new proteins. Defective proteasomal degradation is associated with a variety of clinical conditions, including Alzheimer's disease, Parkinson's disease, Huntington's disease, muscular dystrophy, cardiovascular diseases, and cancer, etc.

[0004] Cereblon is a binding protein of thalidomide and is part of the E3 ubiquitin ligase protein complex. It acts as a substrate receptor to selectively target ubiquitinated proteins. Cereblon is a protein encoded by the human CRBN gene. Cereblon, together with damaged DNA binding protein 1 (DDBl), Cullin-4A (CUL4A), and Cullin-1 regulator (ROCI), forms an E3 ubiquitin ligase complex, which can ubiquitinate a series of proteins, but the specific mechanism is still unclear. Cereblon is currently a commonly used E3 ligase applied to the PROTACs technology.

[0005] The present invention discloses a class of novel spirocyclic compounds, which can be used as effective CRBN ligands, and further can synthesize corresponding PROTACs bifunctional compounds that can make protein degradation targeted chimeras, and can be used to treat various medical conditions, especially abnormal cell proliferation. Summary of the Invention

[0006] The present invention provides a compound represented by Formula I, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt:

[0007]

[0008] Wherein, X1 is selected from CH or N; X2 is selected from CH or N;

[0009] Ring A is selected from 3- to 6-membered saturated or unsaturated monocyclic heteroalkyl; wherein, Ring A may be further optionally substituted by one, two, three or four independent R A1 substituents;

[0010] Each R A1 is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR A2 R A3 , -C 1～6 alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl, halogen-substituted -C 1～6 alkyl, halogen-substituted -C 2～6 alkenyl, halogen-substituted -C 2～6 alkynyl, -C 0～4 alkylene-OR A2 , -C 0～4 alkylene-OC(O)R A2 , -C 0～4 alkylene-SR A2 , -C 0～4 alkylene-S(O)2R A2 , -C 0～4 alkylene-S(O)R A2 , -C 0～4 alkylene-S(O)2NR A2 R A3 , -C 0～4 alkylene-S(O)NR A2 R A3 , -C 0～4 alkylene-C(O)R A2 , -C 0～4 alkylene-C(O)OR A2 , -C 0～4 alkylene-C(O)NR A2 R A3 , -C 0～4 alkylene-NR A2 R A3 , -C 0～4 alkylene-NR A2 C(O)R A3 , -C 0～4 alkylene-NR A2S(O)2R A3 、 -C 0～4 alkylene - NR A2 S(O)R A3 、 -C 0～4 alkylene - (3 - to 10 - membered cycloalkyl), -C 0～4 alkylene - (4 - to 10 - membered heterocycloalkyl), -C 0～4 alkylene - (6 - to 10 - membered aromatic ring), -C 0～4 alkylene - (5 - to 10 - membered heteroaromatic ring); wherein, the alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, and heteroaromatic ring may further optionally be substituted by one, two, three, or four independent R A4 substituents;

[0011] Each R A4 is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR A2 R A3 、 -C 1～6 alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl, halogen - substituted -C 1～6 alkyl, halogen - substituted -C 2～6 alkenyl, halogen - substituted -C 2～6 alkynyl, -C 0～4 alkylene - OR A2 、 -C 0～4 alkylene - OC(O)R A2 、 -C 0～4 alkylene - SR A2 、 -C 0～4 alkylene - S(O)2R A2 、 -C 0～4 alkylene - S(O)R A2 、 -C 0～4 alkylene - S(O)2NR A2 R A3 、 -C 0～4 alkylene - S(O)NR A2 R A3 、 -C 0～4 alkylene - C(O)R A2 、 -C 0～4 alkylene - C(O)OR A2 、 -C 0～4 alkylene - C(O)NR A2 R A3 、 -C 0～4 alkylene - NR A2 R A3 、 -C 0～4 alkylene - NR A2 C(O)R A3 、 -C0～4 Alkylene-NR A2 S(O)2R A3 、-C 0～4 Alkylene-NR A2 S(O)R A3 ;

[0012] R A2 、R A3 are each independently selected from hydrogen, -C 1-6 alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl, halogen-substituted -C 1～6 alkyl, halogen-substituted -C 2～6 alkenyl, halogen-substituted -C 2～6 alkynyl.

[0013] Preferably: the compound is of the structure shown in formula IIA or formula IIB or formula IIC:

[0014]

[0015] is selected from a single bond or a double bond; the R A1 , X1, X2 are as described above.

[0016] Preferably: X1 is selected from CH, X2 is selected from CH; or X1 is selected from N, X2 is selected from CH; or X1 is selected from CH, X2 is selected from N; or X1 is selected from N, X2 is selected from N.

[0017] Preferably: the A ring is a 5-membered nitrogen-containing heterocycle or a 6-membered nitrogen-containing heterocycle.

[0018] Furthermore: the A ring is selected from wherein the ring selected by the A ring may be further optionally substituted by one, two, three or four independent R A1 substituents.

[0019] Preferably: each R A1 is independently selected from hydrogen, halogen, =O, -C 1～3 alkyl, halogen-substituted -C 1～3 alkyl.

[0020] More specifically: the A ring is selected from

[0021] In some specific embodiments of the invention, the compound shown in formula I of the invention is specifically:

[0022]

[0023]

[0024] The present invention also provides the use of the above-mentioned compound, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt in the treatment of diseases related to abnormal cell proliferation.

[0025] Furthermore, the disease is cancer.

[0026] The present invention also provides the use of the above-mentioned compound, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt in the preparation of a targeted protein degradation drug.

[0027] The present invention also provides the use of the above-mentioned compound, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt as an intermediate in the preparation of a targeted protein degradation drug.

[0028] The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0029] Definition of terms used in the present invention: Unless otherwise specified, the initial definition provided for a group or term herein applies to that group or term throughout the specification; for terms not specifically defined herein, meanings should be given based on the disclosure and context that would be understood by those skilled in the art.

[0030] "Substituted" means that a hydrogen atom in a molecule is replaced by other different atoms or molecules. "Substituted" can also mean that a lone pair of electrons of an atom in a molecule is replaced by "=O", "=S", etc.

[0031] "May be further optionally substituted" means that "substitution" can but does not have to occur, and this description includes the cases of occurrence or non-occurrence.

[0032] The minimum and maximum carbon atom contents in a hydrocarbon group are indicated by a prefix. For example, the prefix C a～b alkyl indicates any alkyl group containing "a" to "b" carbon atoms. Thus, for example, "C 1～4 alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0033] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of member atoms. For example, C1-6 alkyl refers to an alkyl group having 1 to 6 member atoms, such as 1 to 4 member atoms. The alkyl group can be straight-chain or branched-chain. Representative branched-chain alkyl groups have one, two, or three branches. The alkyl group may optionally be substituted with one or more substituents as defined herein. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl. The alkyl group can also be part of other groups, such as C1-6 alkoxy.

[0034] As used in the present invention, "alkylene" refers to a divalent saturated aliphatic hydrocarbon group having a specified number of carbon atoms. "C a - b alkylene" refers to an alkylene group having a to b carbon atoms. The alkylene group includes branched-chain and straight-chain hydrocarbon groups. For example, "C 1～6 alkylene" is intended to include methylene, ethylene, propylene, 2-methylpropylene, dimethylethylene, pentylene, and the like. Thus, the term "propylene" can be exemplified by the following structures: Similarly, the term "dimethylbutylene" can be exemplified by any of the following structures: In addition, the term "(C1-6) alkylene" is intended to include such branched-chain hydrocarbon groups, such as cyclopropylmethylene, which can be exemplified by the following structure: For example, -C0-4 alkylene can be C0 alkylene, C1 alkylene (e.g., -CH2-), C2 alkylene (e.g., -CH2CH2-, etc.), C3 alkylene, or C4 alkylene; C0 alkylene means that the group does not exist here and is connected in the form of a chemical bond, such as A-C0 alkylene-B means A-B, that is, group A and group B are directly connected by a chemical bond.

[0035] "Alkenyl" refers to a straight-chain or branched-chain hydrocarbon group having a specified number of carbon atoms and, in some embodiments, 2 to 6 carbon atoms or 2 to 4 carbon atoms and having at least 1 vinyl unsaturation site (>C=C<). For example, C a-b alkenyl refers to an alkenyl group having a to b carbon atoms and is intended to include, for example, vinyl, propenyl, isopropenyl, 1,3-butadienyl, and the like.

[0036] As used in the present invention, "alkenylene" refers to a hydrocarbon chain having 2 to 10 carbon atoms, at least one double bond, and two unsaturated chemical valences. For example, (C3-C6) alkenylene includes >C=CH-CH2-, -CH-CH=CH-CH2-, and the like.

[0037] "Alkynyl" means a straight-chain or branched-chain monovalent hydrocarbon group containing at least one triple bond. The term "alkynyl" also intends to include those hydrocarbon groups having one triple bond and one double bond. For example, (C2-C6) alkynyl intends to include ethynyl, propynyl, etc.

[0038] "Halogen" is fluorine, chlorine, bromine or iodine.

[0039] "Halogenoalkyl", "halogen-substituted alkyl" means that the hydrogen atoms in the alkyl can be substituted by one or more halogens. For example, C 1～4 Halogenoalkyl means an alkyl containing 1 to 4 carbon atoms in which the hydrogen atoms are substituted by one or more halogens.

[0040] "-OR", "-NRR", etc. as described in the present invention mean that the R group is connected to an oxygen atom or a nitrogen atom by a single bond.

[0041] In "-C(O)R", "-S(O)2R", etc. as described in the present invention, the oxygen atom is connected to a carbon atom or a sulfur atom by a double bond, and the R group is connected to the oxygen atom or the sulfur atom by a single bond.

[0042] "Cycloalkyl", "cycloalkane" as described in the present invention means a saturated or partially saturated cyclic group having multiple carbon atoms, no ring heteroatoms, and having a single ring or multiple rings (including fused, bridged, spiro and adamantane systems). For a polycyclic system having aromatic and non-aromatic rings without ring heteroatoms, when the connection point is at a non-aromatic carbon atom, the term "cycloalkyl" is applicable (for example, 5,6,7,8-tetrahydronaphthalen-5-yl). The term "cycloalkyl" includes cycloalkenyl groups, such as cyclohexenyl. Examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl and cyclohexenyl. Examples of cycloalkyl groups including polybicycloalkyl ring systems are dicyclohexyl, dicyclopentyl, dicyclooctyl, etc. For example Adamantyl includes but is not limited to the following structures:

[0043] As used in the present invention, "heterocycle", "heterocycloalkyl", and "heteroalkane" refer to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom; the heteroatom refers to a nitrogen atom, an oxygen atom, a sulfur atom, etc. It generally represents a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of multiple ring atoms, preferably a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of 3 to 9 ring atoms, which contains 1, 2, or 3 ring heteroatoms selected from N, O, and S, and the remaining ring atoms are carbon. A bicyclic ring means composed of two rings sharing two ring atoms, that is, the bridge separating the two rings is a single bond or a chain of one or two ring atoms. Examples of monocyclic saturated heterocycloalkyls are oxetanyl, azetidinyl, pyrrolidinyl, 2-oxo-pyrrolidin-3-yl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azepanyl, diazepanyl, homopiperazinyl or oxazepanyl. Examples of bicyclic saturated heterocycloalkyls are 8-aza-bicyclo[3.2.1]octyl, quinuclidinyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl, 9-aza-bicyclo[3.3.1]nonyl, Examples of partially unsaturated heterocycloalkyls are dihydrofuranyl, imidazolinyl, tetrahydro-pyridyl or dihydropyranyl.

[0044] "Spiroheterocyclic group" and "spiroheterocycle" are used interchangeably, and refer to a non-aromatic saturated ring or a non-aromatic unsaturated ring system in which two monocyclic rings share a carbon atom, which is composed of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus. For example, "5- to 12-membered spiroheterocycle" refers to a spiroheterocycle having 5 to 12 ring atoms, where 1, 2, or 3 ring atoms are heteroatoms.

[0045] "Bridged ring or bridged ring group" refers to a saturated or unsaturated cyclic group formed by two or more cyclic structures sharing two non-adjacent atoms. Specific examples thereof include, but are not limited to:

[0046] "Bridged heterocyclic group" and "bridged heterocycle" are used interchangeably, and refer to a saturated or unsaturated cyclic group formed by two or more cyclic structures sharing two non-adjacent atoms, which is composed of carbon atoms and heteroatoms selected from nitrogen, oxygen, sulfur, and phosphorus. Specific examples thereof include, but are not limited to:

[0047] As used herein, the terms "aromatic ring" and "aryl" refer to aromatic hydrocarbon groups having multiple carbon atoms. An aryl group is typically a monocyclic, bicyclic or tricyclic aryl group having 5 - 20 carbon atoms. In addition, the term "aryl" as used herein refers to an aromatic substituent that can be a single aromatic ring or multiple aromatic rings fused together. Non-limiting examples include phenyl, naphthyl or tetrahydronaphthyl.

[0048] As used herein, the terms "heteroaromatic ring" and "heteroaryl" refer to an aromatic unsaturated ring containing at least one heteroatom; where the heteroatom refers to a nitrogen atom, an oxygen atom, a sulfur atom, etc. It is generally an aromatic monocyclic or bicyclic hydrocarbon containing multiple ring atoms, one or more of which are heteroatoms selected from O, N, S. Preferably, there are one to three heteroatoms. Heteroaryl groups, for example, include: pyridyl, indolyl, quinoxalinyl, quinolinyl, isoquinolinyl, benzothienyl, benzofuranyl, benzothienyl, benzopyranyl, benzothiopyranyl, furyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, oxadiazolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

[0049] "Stereoisomers" include enantiomers and diastereoisomers;

[0050] The "deuterated compound" of the present invention refers to a compound in which one or more hydrogen atoms in a molecule or group are replaced by deuterium atoms, where the proportion of deuterium atoms is greater than the natural abundance of deuterium.

[0051] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or the salt formed is generally chemically or physically compatible with the other components constituting a pharmaceutical dosage form and is physiologically compatible with the receptor.

[0052] The terms "salt" and "pharmaceutically acceptable salt" refer to acid salts and / or base salts formed from the above compounds or their stereoisomers with inorganic and / or organic acids and bases, and also include zwitterionic salts (inner salts), and also include quaternary ammonium salts, such as alkylammonium salts. These salts can be directly obtained in the final separation and purification of the compound. They can also be obtained by appropriately (e.g., equimolarly) mixing the above compounds, or their stereoisomers, with a certain amount of acid or base. These salts may form precipitates in solution and be collected by filtration, or recovered after evaporation of the solvent, or prepared by freeze-drying after reaction in an aqueous medium. The salts described in the present invention can be hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

[0053] Obviously, based on the above content of the present invention, various other modifications, substitutions or alterations can be made without departing from the above basic technical idea of the present invention according to the common technical knowledge and customary means in the art. Detailed implementation manners

[0054] The following is a further detailed description of the above content of the present invention through specific implementation manners in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention.

[0055] The known starting materials of the present invention can be adopted or synthesized according to methods known in the art, or can be purchased from companies such as Aladdin Chemistry, Chengdu Kelong Chemical Industry, Shaoyuan Chemical Technology, J&K Scientific, etc.

[0056] Unless otherwise specified in the examples, the reaction is carried out under a nitrogen atmosphere. Unless otherwise specified in the examples, the solution refers to an aqueous solution. Unless otherwise specified in the examples, the reaction temperature is room temperature. Room temperature is the most suitable reaction temperature, which is 20°C to 30°C. Unless otherwise specified in the examples, M is molar per liter.

[0057] The structures of the compounds are determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). The NMR shift (δ) is given in units of 10-6 (ppm). The NMR measurements are performed using (Bruker AvanceIII 400 and Bruker Avance600) nuclear magnetic resonance spectrometers, and the solvents for measurement are deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (Methol-d4), and the internal standard is tetramethylsilane (TMS). The LC-MS measurements are performed using a Shimadzu liquid chromatography-mass spectrometry instrument (Shimadzu LC-MS2020 (ESI)). The HPLC measurements are performed using a Shimadzu high-performance liquid chromatography instrument (Shimadzu LC-20A). MPLC (medium-pressure preparative chromatography) uses a Gilson GX-281 reversed-phase preparative chromatography instrument. The thin-layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates, and the specifications for separating and purifying products by thin-layer chromatography are 0.4 mm to 0.5 mm. Column chromatography generally uses Yantai Huanghai silica gel with 200 to 300 mesh as the carrier.

[0058] The abbreviations of the reagents described in the examples are as follows: DIPEA: N,N-diisopropylethylamine; EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; HOBT: 1-hydroxybenzotriazole; TCFH: N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate; NMI: N-methylimidazole; Pd(OAc)2: palladium acetate; P( tBu3)BF4: tri-tert-butylphosphine tetrafluoroborate; AcOH: acetic acid; MeOH: methanol; THF: tetrahydrofuran; DMF: N,N-dimethylformamide; DME: ethylene glycol dimethyl ether; DMSO: dimethyl sulfoxide; LC-MS: liquid chromatography-mass spectrometry; MPLC: medium pressure liquid chromatography; HPLC: high pressure liquid chromatography.

[0059] Example 1. Preparation of Compound A1

[0060]

[0061] Step 1. Synthesis of compound 3

[0062] To a 50 mL reaction vial were added compound 1 (200.00 mg, 840.40 μmol), EDCI (321.03 mg, 1.68 mmol), HOBT (227.11 mg, 1.68 mmol), and DMF (5 mL) in sequence. After stirring in an ice bath for 10 minutes, compound 2 (94.35 mg, 1.01 mmol) was added. The reaction was stirred at room temperature for 16 hours, then quenched (LC-MS monitoring). Extraction was completed with saturated NaCl solution (10.0 mL) and ethyl acetate (3 × 10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated to dryness. The product was purified by MPLC and concentrated under reduced pressure to remove the solvent to afford compound 3 (160.00 mg, 510.33 μmol, 60.72% yield).

[0063] Step 2: Preparation of Compound 4

[0064] Compound 3 (150.00 mg, 478.43 μmol), NaH (38.27 mg, 956.87 μmol, 60% dispersion in mineral oil) and DMF (3 mL) were added to a 50 mL reaction bottle. The reaction was stirred at room temperature for 1 hour and then quenched (LC-MS monitoring). The mixture was extracted with saturated NaCl solution (10.0 mL) and ethyl acetate (3×10 mL). The organic phases were combined and dried over anhydrous sodium sulfate. The solvent was evaporated to give compound 4 (100.00 mg, crude product).

[0065] Step 3. Preparation of Compound 5

[0066] Under nitrogen protection, compound 4 (150.00 mg, 541.40 μmol), DME / H2O (4 mL / 1 mL), Mo(CO)6 (214.39 mg, 812.09 μmol), Pd(OAc)2 (12.13 mg, 54.14 μmol), and P( tBu3)BF4 (31.40 mg, 108.28 μmol). Seal the reaction system. After stirring the reaction at 100 °C for 16 hours, quench the reaction (monitored by LC-MS). Extract with saturated NaCl solution (20 mL) and ethyl acetate (3 × 20 mL). Combine the organic phases. Dry the organic phases with anhydrous sodium sulfate, evaporate the solvent, purify by MPLC, and concentrate under reduced pressure to remove the solvent to obtain compound 5 (100.00 mg, 512.36 μmol, 94.64% yield).

[0067] Step 4: Preparation of compound A1

[0068] Add TCFH (4.43 mg, 15.79 μmol), NMI (25.21 mg, 307.42 μmol) and DMF (2 mL) successively into a 50 mL reaction flask. After the temperature of the reaction system drops to 0 °C, add compound 5 (20.00 mg, 102.47 μmol) and compound 6 (14.36 mg, 102.47 μmol). Stir the reaction at room temperature for 1 hour and then quench the reaction (monitored by LC-MS). Extract with saturated NaCl solution (20.0 mL) and dichloromethane (3 × 15 mL). Combine the organic phases. Dry the organic phases with anhydrous sodium sulfate, evaporate the solvent, purify by HPLC, and concentrate under reduced pressure to remove the solvent to obtain compound A1 (3.4 mg, 10.72 μmol, 10.46% yield, purity 90.0%). LC-MS: C 14 H 15 N5O4, [M + H] + 318.11; found 318.1. 1 1H NMR (400 MHz, DMSO-d6) δ 7.67 (s, 1H), 4.34 (t, J = 6.2 Hz, 2H), 4.12 (dd, J = 29.6, 10.0 Hz, 2H), 4.01 - 3.95 (m, 2H), 3.79 - 3.71 (m, 2H), 2.99 (s, 3H), 2.96 - 2.84 (m, 2H).

[0069] Example 2: Preparation of compound B1

[0070]

[0071] Step 1: Synthesis of compound 9

[0072] In a 100 mL reaction flask, compound 7 (5.90 g, 41.52 mmol), compound 8 (2.50 g, 41.52 mmol), and MeOH (15 mL) were added. The reaction was stirred at 0 °C for 8 hours. After the reaction was complete, the reaction was quenched by adding water (monitored by LC-MS). The mixture was extracted with saturated NaCl solution (50 mL) and ethyl acetate (3 × 50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, the solvent was evaporated, and the residue was purified by MPLC. After concentrating under reduced pressure to remove the solvent, compound 9 (700.00 mg, 4.11 mmol, 9.91% yield) was obtained.

[0073] Step 2: Preparation of compound 11

[0074] In a 50 mL reaction flask, compound 9 (100.00 mg, 587.66 μmol), compound 10 (115.81 mg, 587.66 μmol), a mixed solution of 4M HCl and 1,4-dioxane (5 mL), and AcOH (5 mL) were added successively. The reaction was stirred at room temperature for 16 hours. After the reaction was complete, the reaction was quenched by adding water (monitored by LC-MS). The mixture was extracted with saturated NaCl solution (50 mL) and ethyl acetate (3 × 50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, the solvent was evaporated, and the residue was purified by MPLC. After concentrating under reduced pressure to remove the solvent, compound 11 (100.00 mg, 514.97 μmol, 87.63% yield) was obtained.

[0075] Step 3: Preparation of compound 12

[0076] In a 50 mL reaction flask, compound 11 (100.00 mg, 514.97 μmol), LiOH (61.67 mg, 2.57 mmol), and THF / H2O (4 mL / 1 mL) were added successively. The reaction was stirred at room temperature for 6 hours. After the reaction was complete, the reaction was quenched by adding water (monitored by LC-MS). The solvent was evaporated, and the residue was purified by MPLC. After concentrating under reduced pressure to remove the solvent, compound 12 (80.00 mg, 444.05 μmol, 86.23% yield) was obtained.

[0077] Step 4: Preparation of compound A2

[0078] In a 50 mL reaction flask, TCFH (12.00 mg, 333.04 μmol), NMI (79.65 mg, 971.36 μmol) and DMF (2 mL) were added in sequence. After the temperature of the reaction system dropped to 0 °C, compound 12 (50.00 mg, 277.53 μmol) and compound 6 (38.89 mg, 277.53 μmol) were added. After stirring the reaction at room temperature for 16 hours, the reaction was quenched (monitored by LC-MS), and extraction was completed with saturated NaCl solution (20.0 mL) and dichloromethane (3 × 15 mL). The organic phases were combined, dried with anhydrous sodium sulfate, the solvent was evaporated, and after purification by HPLC and concentration under reduced pressure to remove the solvent, compound A2 (4.0 mg, 13.23 μmol, 4.77% yield, purity 97.3%) was obtained. LC-MS: C 14 H 14 N4O4, [M+H] + 303.10; found 303.3. 1 H NMR (400 MHz, DMSO-d6) δ 7.00 (d, J = 2.4 Hz, 1H), 6.25 (d, J = 2.8 Hz, 1H), 4.16 - 4.03 (m, 4H), 4.01 - 3.94 (m, 2H), 3.47 (t, J = 5.6 Hz, 2H), 3.01 - 2.82 (m, 2H).

[0079] The technical effects of the present invention are illustrated by the following test examples:

[0080] Test Example 1: Detection of the inhibition of CRBN / DDB1 activity by the compound (FRET)

[0081] 1. Experimental materials and reagents

[0082] Microplate reader (BMG PHERAstar FSX), ECHO (LABCYTE Echo 665), microplate thermostatic oscillator (Hangzhou Ruicheng Instrument Co., Ltd.), disodium hydrogen phosphate (Sigma), sodium dihydrogen phosphate (Sigma), bovine serum albumin (Sigma), Anti-6His-Tb crypate Gold (CISBIO), CRBN / DDB1 protein (HitGen), 384-well plate (Grenier Bio-one).

[0083] 2. Experimental method

[0084] Dissolve the compound dry powder with DMSO, perform gradient dilution of the compound using ECHO, and add it to a 384-well reaction plate to make the final concentration of DMSO in the whole reaction system (10.0 μL) 1.0%, and add an equal amount of DMSO as a control.

[0085] Dilute the CRBN / DDB1 protein to twice the required final concentration (5.0 nM) using a buffer containing 20 mM disodium hydrogen phosphate, 20 mM sodium dihydrogen phosphate, 0.08% bovine serum albumin, and pH 7.0. Pipette 5.0 μL of the diluted CRBN / DDB1 protein into a 384-well reaction plate that already contains the compound, centrifuge at 1000 rpm for 1 minute, then place on a microplate thermostatic oscillator at 25 °C and 250 rpm for pre-incubation for 15 minutes. Dilute Anti-6His-Tb crypate Gold and the FITC-labeled thalidomide analogue to twice the required final concentration using a buffer containing 20 mM disodium hydrogen phosphate, 20 mM sodium dihydrogen phosphate, 0.08% bovine serum albumin, and pH 7.0. The final concentration of Anti-6His-Tb crypate Gold is 0.2 nM, and the final concentration of the FITC-labeled thalidomide analogue is 50.0 nM to obtain a mixture of Anti-6His-Tb crypate Gold / FITC-labeled thalidomide analogue. Pipette 5.0 μL of the Anti-6His-Tb crypate Gold / FITC-labeled thalidomide analogue mixture into the 384-well reaction plate, centrifuge at 1000 rpm for 1 minute, then place on a microplate thermostatic oscillator at 25 °C and 250 rpm for incubation for 30 minutes. After the reaction, read the fluorescence signal value (Ex = 337 nm, Em = 520 / 490 nm) in the 384-well reaction plate using a microplate reader.

[0086] 3. Data analysis

[0087] Use the solvent group (containing 5.0 nM CRBN / DDB1, 0.2 nM Anti-6His-Tb crypate Gold, 50.0 nM FITC-labeled thalidomide analogue, and 1.0% DMSO) as the negative control and the reaction buffer group (containing 0.2 nM Anti-6His-Tb crypate Gold, 50.0 nM FITC-labeled thalidomide analogue, and 1.0% DMSO) as the blank control;

[0088] Calculate the percentage of residual activity at each concentration using the following formula:

[0089] Residual activity (%) = 100% × (Flu 化合物组 - Flu 空白对照 ) / (Flu 阴性对照 - Flu 空白对照 )

[0090] Then use GraphPad 6.0 to fit the dose-effect curve and calculate the IC 50 value.

[0091] Table 1: Inhibition Table of Compounds and CRBN / DDB1 Proteins

[0092] Number <![CDATA[IC 50 > A1 0.56 μM A2 0.77 μM

Claims

1. A compound represented by Formula I, or a stereoisomer thereof, or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof: in, Said X1 is selected from CH or N; said X2 is selected from CH or N; Ring A is selected from 3 to 6 membered saturated or unsaturated monocyclic heterocyclic alkyl groups; wherein Ring A may be further optionally replaced by one, two, three or four independent R A1 replace; Each R A1 are independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR A2 R A3 、-C 1～6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, halogen-substituted-C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted-C 2～6 Alkynyl, -C 0～4 Alkylene-OR A2 、-C 0～4 Alkylene-OC(O)R A2 、-C 0～4 Alkylene-SR A2 、-C 0～4 Alkylene-S(O)2R A2 、-C 0～4 Alkylene-S(O)R A2 、-C 0～4 Alkylene-S(O)2NR A2 R A3 、-C 0～4 Alkylene-S(O)NR A2 R A3 、-C 0～4 Alkylene-C(O)R A2 、-C 0～4 Alkylene-C(O)OR A2 、-C 0～4 Alkylene-C(O)NR A2 R A3 、-C 0～4 Alkylene-NR A2 R A3 、-C 0～4 Alkylene-NR A2 C(O)R A3 、-C 0～4 Alkylene-NR A2 S(O)2R A3 、-C 0～4 Alkylene-NR A2 S(O)R A3 、-C 0～4 Alkylene-(3-10 membered cycloalkyl), -C 0～4 Alkylene-(4-10 membered heterocycloalkyl), -C 0～4 Alkylene-(6-10 membered aromatic ring), -C 0～4 Alkylene-(5-10 membered heteroaromatic ring); wherein, alkylene, cycloalkyl, heterocycloalkyl, aromatic ring, heteroaromatic ring may be further optionally replaced by one, two, three or four independent R A4 replace; Each R A4 are independently selected from hydrogen, halogen, cyano, nitro, =O, =S, =CR A2 R A3 、-C 1～6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, halogen-substituted-C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted-C 2～6 Alkynyl, -C 0～4 Alkylene-OR A2 、-C 0～4 Alkylene-OC(O)R A2 、-C 0～4 Alkylene-SR A2 、-C 0～4 Alkylene-S(O)2R A2 、-C 0～4 Alkylene-S(O)R A2 、-C 0～4 Alkylene-S(O)2NR A2 R A3 、-C 0～4 Alkylene-S(O)NR A2 R A3 、-C 0～4 Alkylene-C(O)R A2 、-C 0～4 Alkylene-C(O)OR A2 、-C 0～4 Alkylene-C(O)NR A2 R A3 、-C 0～4 Alkylene-NR A2 R A3 、-C 0～4 Alkylene-NR A2 C(O)R A3 、-C 0～4 Alkylene-NR A2 S(O)2R A3 、-C 0～4 Alkylene-NR A2 S(O)R A3 ; R A2 、R A3 are independently selected from hydrogen, -C 1-6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, halogen-substituted-C 1～6 Alkyl, halogen-substituted -C 2～6 Alkenyl, halogen-substituted-C 2～6 Alkynyl.

2. The compound according to claim 1, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt, characterized in that: The compound has a structure shown in Formula IIA, Formula IIB or Formula IIC: is selected from a single bond or a double bond; said R A1 , X1, X2 as described in claim 1.

3. The compound according to claim 1, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt, characterized in that: Said X1 is selected from CH, X2 is selected from CH; or X1 is selected from N, X2 is selected from CH; or X1 is selected from CH, X2 is selected from N; or X1 is selected from N, X2 is selected from N.

4. The compound according to claim 1, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt, characterized in that: The A ring is a 5-membered nitrogen-containing heterocycle or a 6-membered nitrogen-containing heterocycle.

5. The compound according to claim 4, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt, characterized in that: The A ring is selected from Wherein, the ring selected from the group consisting of A and B may be further optionally replaced by one, two, three or four independent R A1 replace.

6. The compound according to claim 5, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt, characterized in that: Each R A1 are independently selected from hydrogen, halogen, =O, -C 1～3 Alkyl, halogen-substituted -C 1～3 alkyl.

7. The compound according to claim 5, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt, characterized in that: The A ring is selected from 8. The compound according to any one of claims 1 to 7, or a stereoisomer thereof, or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof, characterized in that: The compound is specifically:

9. Use of the compound according to any one of claims 1 to 8, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt in the treatment of diseases related to abnormal cell proliferation.

10. The use according to claim 9, characterized in that: The disease is cancer.

11. Use of the compound according to any one of claims 1 to 8, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt in the preparation of a drug for targeted protein degradation.

12. Use of the compound according to any one of claims 1 to 8, or its stereoisomer, or its deuterated compound, or its pharmaceutically acceptable salt as an intermediate in the preparation of a targeted protein degradation drug.

Images