Compound of a hematopoietic progenitor cell kinase 1 inhibitor, its preparation method and application

Abstract

The present invention discloses a compound of a hematopoietic progenitor kinase 1 (HPK1) inhibitor, a preparation method thereof, and an application thereof. Specifically, the present invention provides a compound represented by Formula I, wherein the definitions of each group are as described in the specification. The compound has excellent HPK1 inhibitory activity, and thus can be used in a pharmaceutical composition for treating cancer and other diseases related to HPK activity.

Description

Technical Field

[0001] The present invention relates to a kinase inhibitor, its preparation method and application, and particularly to a compound used as a hematopoietic progenitor kinase 1 (HPK1) inhibitor, its preparation method and application. Background Art

[0002] Surgical resection, radiotherapy, chemotherapy, and small molecule targeted drugs are the main methods for treating cancer. Unfortunately, for many forms of cancer or tumors, surgical resection is often not a viable option, and radiotherapy and chemotherapy can also damage healthy cells while killing tumor cells. In addition, the genomic instability of tumor cells promotes tumor cell mutations, further leading to rapid changes in the cancer genome and making it resistant to drugs specifically targeting tumors, which makes cancer treatment extremely difficult. In recent years, killing tumor cells through the cancer patient's own immune system and enhancing the body's anti-tumor immunity is a new strategy for cancer treatment. One method is to inhibit the negative regulators of the immune response that maintain peripheral tolerance function, so that tumors can be recognized as non-self antigens, thereby overcoming the immune escape of tumor cells. Hematopoietic progenitor kinase (HPK1) is one of the members of the mitogen-activated protein kinase (MAP4K) family, and other members of this family include GCK / MAP4K2, GLK / MAP4K3, HGK / MAP4K4, KHS / MAP4K5, MINK / MAP4K6. HPK1 is a negative regulator of the activation response of B cells, T cells, and dendritic cells. Inhibiting its expression can specifically enhance the body's anti-tumor immunity. It is mainly expressed in hematopoietic cells, such as T cells, B cells, dendritic cells, macrophages, mast cells, and neutrophils. In T cells, HPK1 regulates the activation of T cells through the TCR signaling pathway. After TCR activation, HPK1 interacts with the T cell receptor protein, is phosphorylated by tyrosine kinases Zap70 and Lck, and at the same time phosphorylates the SLP-76 receptor protein, negatively regulating the TCR signal, thereby inhibiting T cell activation and proliferation. Studies have found that HPK1 can participate in many signal cascades, including the MAKP signaling pathway, the Fas-induced apoptosis pathway, and the NF-κB signaling pathway. Moreover, HPK1 can also inhibit AP-1, which plays a role in promoting cell proliferation, inhibiting differentiation, promoting tumor cell invasion and metastasis, etc. during tumor formation and development. HPK1 kinase is not expressed in the main organs, which implies that HPK1 kinase inhibitors may not cause any serious complications.

[0003] Currently, there are no marketed drugs targeting the hematopoietic progenitor kinase (HPK1) target. Summary of the Invention

[0004] OBJECT OF THE INVENTION: The present invention aims to provide a compound having selectivity and high activity and serving as a hematopoietic progenitor kinase 1 inhibitor; another object of the present invention is to provide a preparation method of a compound serving as a hematopoietic progenitor kinase 1 inhibitor; another object of the present invention is to provide the use of a compound serving as a hematopoietic progenitor kinase 1 inhibitor in the preparation of a pharmaceutical composition for preventing or treating a disease responsive to the inhibition of HPK1 activity in a subject.

[0005] TECHNICAL SOLUTION: A compound of the present invention is shown by the following formula I, or a pharmaceutically acceptable salt, isomer or hydrate thereof:

[0006]

[0007] Wherein:

[0008] X is C or N; and when X is C, it may be substituted by R, that is, R is located on X (as CR);

[0009] M 1 、M 2 、M 3 、M 4 or M 5 are each independently CH or N; and when M 1 、M 2 、M 3 、M 4 or M 5 is CH, and may be located on M 1 、M 2 、M 3 、M 4 or M 5 (that is, M 1 、M 2 、M 3 、M 4 or M 5 is C);

[0010] R is H or halogen;

[0011] R 1 and R 2 are each independently selected from the following group: H, deuterium, halogen, OH, CN, NO 2 、C 1-6 deuterated alkyl, C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 6-10 aryl, P(O)R a Rb 、 S(O) 2 R a 、 S(O) 2 NR a R b 、 NR a R b 、 C(O)NR a R b 、 C(O)NR a S(O) 2 R b 、 NR a S(O) 2 R b 、 C(O)R a 、 NR a C(O)R b 、 A 5- to 12-membered heteroaryl having 1 to 3 heteroatoms selected from the group consisting of N, S, and O, or a 3- to 12-membered heterocyclic group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O, wherein the alkyl, alkenyl, alkynyl, alkoxy, phenyl, heteroaryl, cycloalkyl, or heterocyclic group in the groups represented by R 1 and R 2 may be substituted with 1 to 3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 、 C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, NR a R b 、 C(=O)NR a R b 、 C(=O)NR a S(=O) 2 R b 、 P(=O)R a R b 、 S(=O) 2 R a 、 S(=O) 2 NR a R b 、 NR a S(=O) 2 R b ; when the aromatic ring in which each of R 1 or R 2 is located is substituted with a plurality of R 1 or R 2 , two adjacent R 1 or R 2 may form C 3-8Cycloalkyl, 5-12 membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S, O, 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S, O.

[0012] R a , R b are each independently H or C 1-6 alkyl, C 1-6 deuterated alkyl, C 3-8 cycloalkyl, C having 1-3 heteroatoms selected from the group consisting of N, S and O 3-12 heterocyclic group, wherein the alkyl, cycloalkyl, heterocyclic group represented by R 6 or R 7 can be substituted by 1-3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 , C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, C 3-12 heterocyclic group. R a and R b together with the same nitrogen atom or phosphorus atom to which they are attached can form C 3-12 heterocyclic group, which can be substituted by 1-3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 , C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 deuterated alkyl, C 1-6 haloalkoxy, C 3-8 cycloalkyl, C 3-12 heterocyclic group;

[0013] p and q are each independently 0, 1, 2, 3, 4 or 5;

[0014] Furthermore, the compound of formula I has any of the structures shown by the following formula:

[0015]

[0016] Wherein:

[0017] X is C or N; and when the X is C, it can be substituted by R, that is, the R is located on X (as CR);

[0018] M 4 or M 5 is CH or N; and when the M 4 or M 5 is CH, the and may be located on the said M 4 or M 5 above (i.e., M 4 or M 5 is C);

[0019] R is H or halogen;

[0020] R 1 and R 2 are each independently selected from the group consisting of: H, deuterium, halogen, OH, CN, NO 2 , C 1-6 deuterated alkyl, C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 6-10 aryl, P(O)R a R b , S(O) 2 R a , S(O) 2 , NR a R b , NR a R b , C(O)NR a R b , C(O)NR a , S(O) 2 R b , NR a , S(O) 2 R b , C(O)R a , NR a , C(O)R b , 5-12-membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S, O, 3-12-membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S, O, wherein the alkyl, alkenyl, alkynyl, alkoxy, phenyl, heteroaryl, cycloalkyl or heterocyclic group in the groups represented by R 1 and R 2 may be substituted by 1-3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 , C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, NR a R b , C(=O)NR a R b , C(=O)NR a , S(=O) 2 Rb 、P(=O)R a R b 、S(=O) 2 R a 、S(=O) 2 NR a R b 、NR a S(=O) 2 R b ; When R 1 or R 2 each of the aromatic rings to which they are attached is substituted by a plurality of R 1 or R 2 , two R 1 or R 2 in the ortho position may form a C 3-8 cycloalkyl group, a 5- to 12-membered heteroaryl group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O, or a 3- to 12-membered heterocyclic group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O.

[0021] R a 、R b are each independently H or a C 1-6 alkyl group, a C 1-6 deuterated alkyl group, a C 3-8 cycloalkyl group, a C 3-12 heterocyclic group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O, wherein the alkyl group, cycloalkyl group, or heterocyclic group represented by R 6 or R 7 may be substituted by 1 to 3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 、C 1-6 alkyl group, a C 1-6 haloalkyl group, a C 1-6 alkoxy group, a C 3-8 cycloalkyl group, a C 3-12 heterocyclic group. R a and R b together with the same nitrogen or phosphorus atom to which they are attached may form a C 3-12 heterocyclic group, which may be substituted by 1 to 3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 、C 1-6 alkyl group, a C 1-6 haloalkyl group, a C 1-6 alkoxy group, a C 1-6 deuterated alkyl group, a C 1-6 haloalkoxy group, a C 3-8 cycloalkyl group, a C 3-12 heterocyclic group;

[0022] p and q are each independently 0, 1, 2, 3, 4, or 5;

[0023] Furthermore, the compound of formula I has any of the structures shown in the following formula:

[0024]

[0025]

[0026] The synthesis scheme is as follows:

[0027]

[0028] Step I: The compound reacts with TsCl under the action of NaH to prepare compound A-1;

[0029] Step II: Compound A-1 reacts with under a metal catalyst such as [PdCl 2 (dppf)]CH 2 Cl 2 to introduce through a Suzuki coupling reaction to obtain compound A-2;

[0030] Step III: Compound A-2 reacts with under a metal catalyst such as [PdCl 2 (dppf)]CH 2 Cl 2 to introduce through a Suzuki coupling reaction to obtain compound A-3;

[0031] Step IV: Compound A-3 is deprotected by Ts under the action of a strong base NaOH to obtain A-4 (i.e., the compound of formula I).

[0032] On the other hand, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of one or more of the above-mentioned compounds, their pharmaceutically acceptable salts, isomers or hydrates, and a pharmaceutically acceptable excipient.

[0033] On the other hand, the present invention provides the use of the above-mentioned compound, or its pharmaceutically acceptable salt, isomer or hydrate in the preparation of a pharmaceutical composition for preventing or treating a disease responsive to the inhibition of HPK1 activity in a subject.

[0034] A combination use scheme of the above-mentioned compound, or its pharmaceutically acceptable salt, isomer or hydrate with other tumor immunotherapeutic agents, wherein the other tumor immunomodulators are selected from the group consisting of: small molecule compounds and antibodies (including but not limited to PD-1, PD-L1, CTLA-4, STING agonists, LAG3 antagonists, etc.), tumor targeting drugs, tumor vaccines, radiotherapy regimens.

[0035] Use of the above-mentioned compound and its pharmaceutically acceptable salts, stereoisomers, prodrugs, solvates, esters and deuterated compounds in combination with CAR-T immunotherapy in cancer immunotherapy.

[0036] Furthermore, the disease is cancer.

[0037] On the other hand, the present invention provides use of the above-mentioned compound, or its pharmaceutically acceptable salt, isomer or hydrate in an HPK1 kinase inhibitor.

[0038] Since the compound of the present invention has very good inhibitory activity against HPK1 kinase, the compound of the present invention and its various crystal forms, pharmaceutically acceptable organic or inorganic salts, solvates or hydrates, and pharmaceutical compositions containing the compound of the present invention as the main active ingredient can be used for preventing and / or treating diseases related to HPK1 kinase activity or expression level (such as, cancer).

[0039] The pharmaceutical composition of the present invention comprises the compound of the present invention within a safe and effective amount range and a pharmaceutically acceptable carrier or excipient. The "safe and effective amount" herein refers to: an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1 - 2000 mg of the compound of the present invention per dose, and preferably, contains 10 - 200 mg of the compound of the present invention per dose. Preferably, the "per dose" is a capsule or a tablet.

[0040] "Pharmaceutically acceptable carrier" refers to: one or more compatible liquid or solid fillers or gelling substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" herein means that the components in the composition can be mixed with the compound of the present invention and with each other, and do not significantly reduce the efficacy of the compound. Some examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium ethylcellulose, sodium carboxymethylcellulose, cellulose acetate, etc.), gelatin, talc, calcium sulfate, solid lubricants (such as stearic acid, magnesium stearate), vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), coloring agents, polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween R, wetting agents (such as sodium dodecyl sulfate), flavoring agents, preservatives, stabilizers, antioxidants, pyrogen-free water, etc.

[0041] There is no particular limitation on the administration mode of the compound or pharmaceutical composition of the present invention. Representative administration modes include (but are not limited to): oral administration, parenteral (intravenous, intramuscular or subcutaneous).

[0042] Solid dosage forms for oral administration include capsules, pills, tablets, powders, and granules. In these solid dosage forms, the active compound is admixed with at least one conventional inert excipient (or carrier), such as dibasic calcium phosphate or sodium citrate, or with the following components: (a) fillers or bulking agents, e.g., lactose, sucrose, starch, glucose, mannitol, and silicic acid; (b) binders, e.g., hydroxypropylmethyl cellulose, gelatin, alginates, polyvinylpyrrolidone, acacia, and sucrose; (c) humectants, e.g., glycerin; (d) disintegrants, e.g., calcium carbonate, agar, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, e.g., paraffin wax; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (h) adsorbents, e.g., kaolin; (i) lubricants, e.g., talc, solid polyethylene glycols, calcium stearate, magnesium stearate, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0043] Solid dosage forms such as tablets, capsules, dragees, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed and released in a portion of the digestive tract. Examples of embedding components that can be used are polymeric and wax-like substances. If necessary, the active compound can also be in the form of microcapsules with one or more of the above excipients.

[0044] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, suspensions, solutions, syrups, or tinctures. In addition to the active compound, the liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizing agents, and emulsifying agents, e.g., ethanol, isopropanol, propylene glycol, ethyl acetate, ethyl carbonate, 1,3-butanediol, dimethylformamide, and oils, especially peanut oil, cottonseed oil, corn germ oil, olive oil, sesame oil, and castor oil, or mixtures of these substances.

[0045] In addition to these inert diluents, the compositions may also contain adjuvants such as emulsifying agents, wetting agents, and suspending agents, flavoring agents, sweetening agents, and perfumes.

[0046] In addition to the active compound, the suspension may contain suspending agents, e.g., polyoxyethylene sorbitol and sorbitan esters, ethoxylated isostearyl alcohols, microcrystalline cellulose, agar, and aluminum monostearate, or mixtures of these substances.

[0047] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or dispersion liquids, non-aqueous solutions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and diluents, non-aqueous carriers, solvents or excipients include water, polyols, ethanol and their suitable mixtures.

[0048] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0049] When administered in combination, the pharmaceutical composition further comprises one or more other pharmaceutically acceptable compounds. One or more of the other pharmaceutically acceptable compounds can be administered simultaneously with, separately from, or sequentially to the compound of the present invention.

[0050] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal in need of treatment (such as a human), wherein the dosage during administration is an effective dosage considered pharmaceutically. For a person weighing 60 kg, the daily dosage is usually 1 - 2000 mg, preferably 20 - 500 mg. Of course, the specific dosage should also consider factors such as the patient's health condition and the administration route, which are within the scope of the skills of a skilled physician.

[0051] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0052] Advantageous effects: Compared with the prior art, the present invention has the following remarkable advantages: It has excellent HPK1 inhibitory activity and can be used to prepare pharmaceutical compositions for treating cancer and other diseases related to HPK activity. Detailed Description of the Invention

[0053] The present invention provides a compound represented by the following formula I:

[0054]

[0055] Wherein:

[0056] X is C or N; and when X is C, it can be substituted by R, that is, R is located on X (as CR);

[0057] M 1 、M 2 、M 3 、M 4 or M 5 are each independently CH or N; and when M 1 、M2 , M 3 , M 4 or M 5 When it is CH, the and can be located on the said M 1 , M 2 , M 3 , M 4 or M 5 above (i.e., M 1 , M 2 , M 3 , M 4 or M 5 is C);

[0058] R is H or halogen;

[0059] R 1 and R 2 are each independently selected from the group consisting of: H, deuterium, halogen, OH, CN, NO 2 , C 1-6 deuterated alkyl, C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 6-10 aryl, P(O)R a R b , S(O) 2 R a , S(O) 2 , NR a R b , NR a R b , C(O)NR a R b , C(O)NR a , S(O) 2 R b , NR a , S(O) 2 R b , C(O)R a , NR a C(O)R b , a 5-12-membered heteroaryl having 1-3 heteroatoms selected from the group consisting of N, S, O, a 3-12-membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S, O, wherein the alkyl, alkenyl, alkynyl, alkoxy, phenyl, heteroaryl, cycloalkyl or heterocyclic group in the groups represented by R 1 and R 2 can be substituted by 1-3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 , C1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, NR a R b , C(=O)NR a R b , C(=O)NR a S(=O) 2 R b , P(=O)R a R b , S(=O) 2 R a , S(=O) 2 NR a R b , NR a S(=O) 2 R b ; when R 1 or R 2 each of the aromatic rings in which they are located is substituted by a plurality of R 1 or R 2 , two adjacent R 1 or R 2 can form C 3-8 cycloalkyl, a 5- to 12-membered heteroaryl having 1 to 3 heteroatoms selected from the group consisting of N, S, and O, or a 3- to 12-membered heterocyclic group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O.

[0060] R a , R b are each independently H or C 1-6 alkyl, C 1-6 deuterated alkyl, C 3-8 cycloalkyl, a C 3-12 heterocyclic group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O, wherein the alkyl, cycloalkyl, or heterocyclic group represented by R 6 or R 7 can be substituted by 1 to 3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 , C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 3-8 cycloalkyl, C 3-12 heterocyclic group. R a and R b together with the same nitrogen or phosphorus atom to which they are attached can form a C 3-12 heterocyclic group, which can be substituted by 1 to 3 substituents independently selected from the group consisting of: halogen, OH, CN, NO 2 , C1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Deuterated alkyl, C 1-6 Haloalkoxy, C 3-8 Cycloalkyl, C 3-12 Heterocyclic group;

[0061] p and q are each independently 0, 1, 2, 3, 4 or 5;

[0062] In the compound of formula I, each chiral center is in the R configuration or the S configuration.

[0063] Preferably, the X, M 1 , M 2 , M 3 , M 4 , M 5 , R, R 1 , R 2 , R a , R b , p, q are each independently the corresponding groups in the specific compounds in each embodiment.

[0064] The compounds of the present invention can be used as HPK1 kinase inhibitors, and in a preferred embodiment, they are HPK1 kinase selective inhibitors.

[0065] Preparation of the compound of formula I

[0066] The compound of formula I of the present invention can be prepared by the following exemplary methods:

[0067]

[0068] Step I: Compound React with TsCl under the action of NaH to prepare compound A-1;

[0069] Step II: Compound A-1 reacts with under a metal catalyst such as [PdCl 2 (dppf)]CH 2 Cl 2 to introduce through a Suzuki coupling reaction to obtain compound A-2;

[0070] Step III: Compound A-2 reacts with under a metal catalyst such as [PdCl 2 (dppf)]CH 2 Cl 2 to introduce through a Suzuki coupling reaction to obtain compound A-3;

[0071] Step IV: Under the action of strong base NaOH, compound A-3 removes the Ts protecting group to obtain A-4 (i.e., the compound of formula I).

[0072] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight.

[0073] Preparation of Key Intermediate 1. 5-Bromo-3-iodo-1-(p-toluenesulfonyl)pyrrolo[2,3-b]pyridine

[0074]

[0075] At 25 °C, NaH (1.36 g, 33.91 mmol) was added to a solution of 5-bromo-3-iodo-1H-pyrrolo[2,3-b]pyridine (7.3 g, 22.61 mmol) in tetrahydrofuran (140 mL). The mixture was stirred at 25 °C for 0.5 h. Then, p-toluenesulfonyl chloride (4.74 g, 24.87 mmol) was added, and the mixture was stirred at 25 °C overnight. After the reaction was completed, it was diluted with saturated NaHCO 3 (100 mL), and extracted with ethyl acetate (3 × 250 mL). The combined extracts were washed with brine (3 × 100 mL), dried over Na 2 SO 4 , filtered and concentrated under reduced pressure to obtain 5-bromo-3-iodo-1-(p-toluenesulfonyl)pyrrolo[2,3-b]pyridine (10.3 g, 19.37 mmol, 86%), as a yellow solid. MS (ESI): M / Z = 478 [M+H] + .

[0076] Preparation of Key Intermediate 2. 5-Bromo-4-chloro-3-iodo-1-toluenesulfonyl-1H-pyrrolo[2,3-b]pyridine

[0077]

[0078] The same procedure as in Key Intermediate 1 was followed to prepare 5-bromo-4-chloro-3-iodo-1-toluenesulfonyl-1H-pyrrolo[2,3-b]pyridine. MS (ESI): M / Z = 512 [M+H] + .

[0079] Preparation of Key Intermediate 3. 2-Bromo-7-iodo-5-toluenesulfonyl-5H-pyrrolo[2,3-b]pyrazine

[0080]

[0081] The procedure is the same as that for the key intermediate 1. 2-Bromo-7-iodo-5-tosyl-5H-pyrrolo[2,3-b]pyrazine was prepared. MS(ESI): M / Z = 479 [M+H] + 。

[0082] Preparation of Key Intermediate 4. 6-Amino-2-fluoro-N,N-dimethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

[0083]

[0084] Step 1: Triethylamine (557 mg, 5.5 mmol) and HATU (1.7 g, 5.5 mmol) were successively added to a suspension of 6-amino-3-bromo-2-fluorobenzoic acid (1.17 g, 5 mmol) in anhydrous dichloromethane (20 mL), and the mixture was stirred for 5 minutes. Then, dimethylamine (2 M in tetrahydrofuran, 5.0 mL) was added at 0 °C. The mixture was stirred at room temperature for 3 hours and then washed with 1 M sodium hydroxide and brine. The aqueous phase was extracted twice more with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate. After filtration by suction, the solvent was removed under reduced pressure to obtain 6-amino-3-bromo-2-fluoro-N,N-dimethylbenzamide without further purification. MS(ESI): M / Z = 183 [M+H] + 。

[0085] Step 2: 6-Amino-3-bromo-2-fluoro-N,N-dimethylbenzamide (2.95 g, 11.3 mmol), bis(pinacolato)diboron (5.74 g, 22.6 mmol), Pd(dppf)Cl 2 (922.8 mg, 1.13 mmol), and AcOK (3.33 g, 33.9 mmol) were added to a reaction flask containing dioxane (25 ml). The reaction was carried out at 85 °C for 3 h under nitrogen protection. The cooled mixture was filtered through Celite, washed thoroughly with ethyl acetate, and the filtrate was evaporated to a small volume. The residue was dissolved in ethyl acetate and water, filtered through Celite again, and the phases were separated. The aqueous phase was extracted 3 times with ethyl acetate. The organic phase was concentrated under reduced pressure. The residue was chromatographed on silica gel, eluting with 50 - 100% ethyl acetate / petroleum ether to obtain 1.81 g of 6-amino-2-fluoro-N,N-dimethyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide. MS(ESI): M / Z = 309 [M+H] +Preparation of Key Intermediate 5. 2-Amino-N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

[0086]

[0087] Steps 1 and 2: Following the same operating procedure as for Key Intermediate 4, 2-Amino-N,N-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide was prepared. MS(ESI): M / Z = 291 [M+H] + 。

[0088] Key Intermediate 6: Preparation of N,N-Dimethyl-2-(4-methylpiperazin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide

[0089]

[0090] Step 1: Following the same operating procedure as Step 1 of Key Intermediate 4, 5-Bromo-2-iodo-N,N-dimethylbenzamide was prepared. MS(ESI): M / Z = 354 [M+H] + 。

[0091] Step 2: Dissolve the 5-Bromo-2-iodo-N,N-dimethylbenzamide (353 mg, 1 mmol) obtained in the previous step in dioxane (4 ml), add N-methylpiperazine (101 mmol, 1 mmol), Pd 2 (dba) 3 (91.6 mg, 0.1 mmol), Xantphos (115.7 mg, 0.2 mmol), cesium carbonate (977 mg, 3 mmol), and reflux the reaction mixture at 110 °C for 24 h under nitrogen protection. Cool to room temperature, filter through diatomaceous earth, rotary evaporate the filtrate under reduced pressure, and purify by column chromatography to obtain 5-Bromo-N,N-dimethyl-2-(4-methylpiperazin-1-yl)benzamide (130 mg). MS(ESI): M / Z = 327 [M+H] + 。

[0092] Step 3: Following the same operating procedure as Step 2 of Key Intermediate 4, N,N-Dimethyl-2-(4-methylpiperazin-1-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide was prepared. MS(ESI): M / Z = 374 [M+H] + 。Preparation of Key Intermediate 7. 2-Methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinoline

[0093]

[0094] Following the operation procedure of Step 2 of Key Intermediate 4, 2-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinoline was finally prepared. MS(ESI): M / Z = 274 [M+H]+.

[0095] Preparation of Key Intermediate 8. 5-Methoxy-2-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinoline

[0096]

[0097] Following the operation procedure of Step 2 of Key Intermediate 4, 5-methoxy-2-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,4-tetrahydroisoquinoline was prepared. MS(ESI): M / Z = 304 [M+H] + 。

[0098] Preparation of Key Intermediate 9. 4-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)morpholine

[0099]

[0100] Following the operation procedure of Step 2 of Key Intermediate 4, 4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)morpholine was prepared. MS(ESI): M / Z = 290 [M+H] + 。

[0101] Preparation of Key Intermediate 10. 1-Methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine

[0102]

[0103] Following the operation procedure of Step 2 of Key Intermediate 4, 1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)piperazine was prepared. MS(ESI): M / Z = 303 [M+H] + 。

[0104] Preparation of Key Intermediate 11. 2,6-Difluoro-N,N-dimethyl-3-nitrobenzamide

[0105]

[0106] Step 1: Follow the operation procedure of Step 1 of the key intermediate 4 to prepare 2,6-difluoro-N,N-dimethyl-3-nitrobenzamide. MS(ESI): M / Z = 231 [M+H] + 。

[0107] Step 2: Add 2,6-difluoro-N,N-dimethyl-3-nitrobenzamide (2.31 g, 10 mmol), N-methylpiperazine (900 mg, 9 mmol), and TEA (2.7 g, 27 mmol) obtained in the previous step to DMSO (20 mL), stir at 100 °C for 2 hours, and cool to room temperature. Dilute the reaction mixture with water (50 mL) and extract with ethyl acetate (3 x 50 mL). Wash the combined organic layers with brine (2 x 50 mL), dry over anhydrous sodium sulfate, and concentrate in vacuo. Purify the residue by silica gel column chromatography to obtain the target compound 2-fluoro-N,N-dimethyl-6-(4-methylpiperazin-1-yl)-3-nitrobenzamide (2.2 g, 78.8%). MS(ESI): M / Z = 311 [M+H] + 。

[0108] Step 3: Place 2-fluoro-N,N-dimethyl-6-(4-methylpiperazin-1-yl)-3-nitrobenzamide (2.2 g, 7.1 mmol), Fe (2 g, 35.5 mmol), 10 drops of concentrated hydrochloric acid, and EtOH (20 mL) / H 2 O (4 mL) into a 40-mL round-bottom flask. Stir the resulting solution at 60 °C for 4 hours and cool to room temperature. Filter through diatomaceous earth, dilute with saturated sodium bicarbonate (50 mL), and extract with ethyl acetate (3 x 60 mL). Dry the combined organic layers and concentrate in vacuo. Purify the residue by silica gel column chromatography to obtain the target compound 3-amino-2-fluoro-N,N-dimethyl-6-(4-methylpiperazin-1-yl)benzamide (1.8 g, 90.4%). MS(ESI): M / Z = 281 [M+H] + 。

[0109] Step 4: Place CuBr into a 40-mL round-bottom flask 2(1.31 g, 5.85 mmol), n-butyl nitrite (482 mg, 4.68 mmol) and ACN (20 mL). The resulting solution was stirred at 0 °C for 5 minutes and treated dropwise at 0 °C with a solution of 3-amino-2-fluoro-N,N-dimethyl-6-(4-methylpiperazin-1-yl)benzamide (655 mg, 2.34 mmol) in ACN (5 mL). The resulting solution was stirred at 50 °C for 1 hour. The reaction mixture was cooled to room temperature and diluted with water (50 mL). The resulting solution was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were dried and concentrated in vacuo. The residue was purified by silica gel column chromatography to give the target compound (201 mg, 35.2%). MS (ESI): M / Z = 345 [M+H] + 。

[0110] Step 5: Following the same procedure as in Step 2 of Key Intermediate 4, 2-fluoro-N,N-dimethyl-6-(4-methylpiperazin-1-yl)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzamide was prepared. MS (ESI): M / Z = 392 [M+H] + Example 1. Preparation of 6-amino-2-fluoro-N,N-dimethyl-3-(3-(pyridin-3-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridin-5-yl)benzamide

[0111]

[0112] Step 1: At room temperature, Key Intermediate 1 (954 mg, 2 mmol), pyridin-3-boronic acid (135.2 mg, 1.1 mmol), Pd(dppf)Cl 2 (146.2 mg, 0.2 mmol), and sodium carbonate (318 mg, 3 mmol) were added to 1,4-dioxane (7 mL) and water (1 mL), and then under N 2 protection conditions, the reaction was carried out at 80 °C for 3 h and the reaction was stopped. The reaction solution was filtered through diatomaceous earth, 20 mL of H 2 O and 20 mL of EA were added to the reaction solution, and then extracted three times with EA. The combined organic phases were washed with saturated sodium chloride and separated by column chromatography to obtain the target compound 5-bromo-3-(pyridin-3-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridine as a white solid (390 mg, 45.5%). MS (ESI): M / Z = 429 [M+H] + 。

[0113] Step 2: At room temperature, add the compound 5-bromo-3-(pyridin-3-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridine (214 mg, 0.5 mmol) obtained in the previous step, key intermediate 4 (169 mg, 0.55 mmol), Pd(dppf)Cl 2 (73 mg, 0.1 mmol), and sodium carbonate (159 mg, 1.5 mmol) into 1,4-dioxane (7 mL) and water (1 mL), then under N 2 protection conditions, react at 80 °C for 3 h and stop the reaction. Filter the reaction solution through diatomaceous earth, add 20 mL of H 2 O and 20 mL of DCM to the reaction solution, then extract with DCM three times, combine the organic phases, wash with saturated sodium chloride, and separate by column chromatography to obtain the target compound 6-amino-2-fluoro-N,N-dimethyl-3-(3-(pyridin-3-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridin-5-yl)benzamide (120 mg, 45.5%) as a white solid. MS(ESI): M / Z = 530 [M+H] + .

[0114] Step 3: Dissolve the 6-amino-2-fluoro-N,N-dimethyl-3-(3-(pyridin-3-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridin-5-yl)benzamide (100 mg, 0.189 mmol) obtained in the previous step in a mixed solution of acetone (4 ml) and methanol (4 ml), add an aqueous solution of sodium hydroxide (2 M, 3 ml), react at 60 °C for 5 h, cool to room temperature, add 20 mL of H 2 O and 20 mL of DCM to the reaction solution, then extract with DCM three times, combine the organic phases, wash with saturated sodium chloride, and separate by column chromatography to obtain the target compound 6-amino-2-fluoro-N,N-dimethyl-3-(3-(pyridin-3-yl)-1-tosyl-1H-pyrrolo[2,3-b]pyridin-5-yl)benzamide (36 mg, 50.8%) as a white solid. MS(ESI): M / Z = 376 [M+H] + . 1 HNMR(400MHz, DMSO-d 6)δ 8.87 (t, J = 1.9 Hz, 1H), 8.73 (dd, J = 1.5, 2.0 Hz, 1H), 8.63 (dt, J = 1.8, 4.7 Hz, 1H), 8.21 (dd, J = 1.5, 2.1 Hz, 1H), 7.96 (dt, J = 1.9, 8.3 Hz, 1H), 7.57 (dd, J = 5.0, 8.3 Hz, 1H), 7.44 (dd, J = 4.8, 8.3 Hz, 1H), 7.14 (d, J = 2.6 Hz, 1H), 6.75 (d, J = 8.4 Hz, 1H), 6.05 (s, 2H), 3.01 (s, 6H).

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[0140] Comparative Example

[0141] The compounds described in Patent CN201980038718.0 were used as comparative examples, and the selected compounds are as follows:

[0142] Comparative Example 1: Comparative Example 2: Comparative Example 3: Biological Activity Test Example 1 HPK1 ADP-Glo Enzymatic Activity Test

[0143] Prepare the enzyme activity test buffer containing 40 mM Tris, pH 7.5; 20 mM MgC12; 0.1 mg / ml BSA; 50 μM DTT. The compound was dissolved in pure DMSO, and the stock solution concentration was 10 mM. The DMSO solution of the compound was serially diluted three-fold starting from 100 μM, with a total of 11 concentrations. The diluted compound was diluted 1:20 with the enzyme activity test buffer, and 1 μl was taken and added to the working wells, with two replicates for each concentration. The negative control well and the positive control well were both added with 1 μl of DMSO solution diluted 1:20. Prepare the 2.5× substrate / ATP working solution as the enzyme activity test buffer containing 45 μM ATP and 0.25 μg / μl MBP protein, and 2 μl of the 2.5× substrate / ATP working solution was added to each working well. Prepare the 2.5× enzyme reaction working solution as the enzyme activity test buffer containing 0.5 ng / μl HPK1 recombinant protein (Signalchem, catalog number M23-11G-10), and 2 μl of the 2.5× enzyme reaction working solution was added to each working well. The negative control well was only added with 2 μl of the enzyme activity test buffer. Stick on the sealing film and centrifuge, and place it at room temperature for reaction for 30 minutes. After the reaction, 5 μl of ADP-Glo reagent (Promega, catalog number V1901) was added to each well and continued to react at room temperature for 40 minutes. 10 μl of kinase detection reagent (Promega, catalog number V1901) was added and reacted at room temperature for 20 minutes, and then the final luminescence signal was measured.

[0144] Calculate the average values of the positive wells and negative wells respectively as the positive control value (Signalpos) and the negative control value (Signalneg). Calculate the inhibition rate of the working well signal value (Signaltest) according to the formula Inhibition rate = (Signalpos - Signaltest) / (Signalpos - Signalneg) × 100%. Plot the concentration-inhibition rate curve by non-linear fitting in GraphPad Prism software for the calculated inhibition rate and calculate the IC50 value.

[0145] Biological Activity Test Example 2 GLK ADP-Glo Enzymatic Activity Test

[0146] Prepare the enzyme activity test buffer containing 40 mM Tris, pH 7.5; 20 mM MgC12; 0.1 mg / ml BSA; 50 μM DTT. Dissolve the compound in pure DMSO with a stock solution concentration of 10 mM. Continuously perform three-fold serial dilutions of the compound DMSO solution starting from 100 μM, with a total of 11 concentrations. Dilute the diluted compound 1:20 with the enzyme activity test buffer, take 1 μl and add it to the working wells, with two replicates for each concentration. Add 1 μl of the 1:20 diluted DMSO solution to both the negative control well and the positive control well. Prepare the 2.5× substrate / ATP working solution as the enzyme activity test buffer containing 105 μM ATP and 0.5 μg / μl PKA substrate polypeptide, and add 2 μl of the 2.5× substrate / ATP working solution to each working well. Prepare the 2.5× enzyme reaction working solution as the enzyme activity test buffer containing 2.5 ng / μl GLK recombinant protein (Signalchem, catalog number M25-11G-10), and add 2 μl of the 2.5× enzyme reaction working solution to each working well. Only add 2 μl of the enzyme activity test buffer to the negative control well. Stick on the sealing film and centrifuge, place it at room temperature for reaction for 1 hour. After the reaction, add 5 μl of ADP-Glo reagent (Promega, catalog number V1901) to each well and react at room temperature for 40 minutes, then add 10 μl of kinase detection reagent (Promega, catalog number V1901) and react at room temperature for 20 minutes, and measure the final luminescence signal.

[0147] Calculate the average values of the positive wells and negative wells respectively as the positive control value (Signalpos) and the negative control value (Signalneg). Calculate the inhibition rate of the working well signal value (Signaltest) according to the formula Inhibition rate = (Signalpos - Signaltest) / (Signalpos - Signalneg) × 100%. Plot the concentration-inhibition rate curve by non-linear fitting in GraphPad Prism software for the calculated inhibition rate and calculate the IC50 value. The experimental results are shown in the following table:

[0148] Table 1 Experimental Results of HPK1 and GLK Enzyme Activity Tests

[0149]

[0150]

[0151] Among them, A represents IC 50 value ≤ 50 nM; B represents 50 nM ≤ IC 50 value ≤ 500 nM; C represents 500 nM ≤ IC 50 value ≤ 10 μM.

Claims

1. A compound represented by the following formula I, or a pharmaceutically acceptable salt thereof, The compound of formula I has any of the structures shown by the following formula:

2. A method for preparing the compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: The method comprises the following steps: Step I: Compound Compound A-1 was prepared by reacting with TsCl under the action of NaH; Step II: Compound A-1 reacts with in the presence of a metal catalyst [PdCl 2 (dppf)]CH 2 Cl 2 to introduce through Suzuki coupling reaction to obtain Compound A-2; Step III: Compound A-2 reacts with in the presence of a metal catalyst [PdCl 2 (dppf)]CH 2 Cl 2 to introduce through a Suzuki coupling reaction to obtain Compound A-3; Step IV: Compound A-3 is deprotected from the Ts protecting group under the action of strong base NaOH to obtain A-4, namely the compound of formula I.

3. A pharmaceutical composition comprising a therapeutically effective amount of one or more of the compounds according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

4. Use of the compound according to claim 1 and a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating a disease responsive to inhibition of HPK1 activity in a subject.

5. The use according to claim 4, characterized in that the disease is cancer.

6. Use of the compound according to claim 1 and a pharmaceutically acceptable salt thereof in the preparation of an HPK1 activity inhibitor.

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