CN115772156B - Compound used as HPK1 kinase inhibitor and preparation method and application thereof - Google Patents

Abstract

The present invention discloses a compound used as an HPK1 kinase inhibitor, a preparation method and an application thereof. Specifically, the present invention provides a compound as shown in 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 as a pharmaceutical composition for treating cancer and other HPK activity-related diseases.

Description

Compound used as HPK1 kinase inhibitor and preparation method and application thereof

Technical Field

The present invention relates to a kinase inhibitor and a preparation method and application thereof, and in particular to a compound used as an HPK1 kinase inhibitor and a preparation method and application thereof.

Background technique

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 feasible option, and radiotherapy and chemotherapy can damage healthy cells while killing tumor cells. In addition, the instability of the tumor cell genome promotes the mutation of tumor cells, further leading to rapid changes in the cancer genome, making it resistant to drugs that specifically target tumors, which makes cancer treatment difficult. In recent years, killing tumor cells through the cancer patient's own immune system and improving the body's anti-tumor immunity is a new strategy for cancer treatment. One of the methods is to inhibit the negative regulators of the immune response that maintains peripheral tolerance, so that the tumor is recognized as a non-self antigen, thereby overcoming the immune escape of tumor cells. Hematopoietic progenitor kinase (HPK1) is a member of the mitogen-activated protein kinase (MAP4K) family, which also includes GCK/MAP4K2, GLK/MAP4K3, HGK/MAP4K4, KHS/MAP4K5, and MINK/MAP4K6. HPK1 is a negative regulator of B cell, T cell, and dendritic cell activation reactions. Inhibiting its expression can specifically improve 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 T cell activation through the TCR signaling pathway. After TCR activation, HPK1 interacts with T cell receptor proteins and is phosphorylated by tyrosine kinases Zap70 and Lck. It also phosphorylates SLP-76 receptor proteins, negatively regulating TCR signals, 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. In addition, HPK1 can also inhibit AP-1, which plays a role in promoting cell proliferation, inhibiting differentiation, and promoting tumor cell invasion and metastasis in tumor formation and development. HPK1 kinase is not expressed in major organs, suggesting that HPK1 kinase inhibitors may not cause any serious complications.

Currently, there is no marketed drug targeting the hematopoietic progenitor cell kinase (HPK1) target.

Summary of the invention

Purpose of the invention: The present invention aims to provide a compound used as a HPK1 kinase inhibitor with selectivity and high activity; another purpose of the present invention is to provide a method for preparing a compound used as a HPK1 kinase inhibitor; another purpose of the present invention is to provide the use of a compound used as a HPK1 kinase inhibitor in the preparation of a pharmaceutical composition for preventing or treating a disease responsive to inhibition of HPK1 activity in a subject.

Technical solution: A compound of the present invention is represented by the following formula (I) or a pharmaceutically acceptable salt, isomer or hydrate thereof:

in:

X ₁ is selected from the group consisting of none, NR, O, S(O) _t , CHR, NRC(O), C(O)NR, NRC(O)C(O)NR, NRC(O)NR, NRC(S)NR, NRC(O)NRCH ₂ , NRC(S)NRCH ₂ , wherein R is selected from the group consisting of H, C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, or a 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O;

_X2 or _X3 are each independently selected from none, NR _a , O, S(O) _t , C(O), CR ₄ R ₅ ;

Y is selected from NR _a , O, S(O) _t or CR ₄ R ₅ ;

M ¹ , M ² , or M ³ are each independently CH or N; and when M ¹ , M ² , or M ³ is CH, may be located on the M ¹ , M ² , or M ³ (ie, M ¹ , M ² , or M ³ is C);

The A ring is selected from the following group: H, C _3-8 cycloalkyl, 3-12 membered heterocyclyl, C _6-10 aromatic ring, 5-10 membered aromatic heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered aromatic ring and a 5-7 membered heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered aromatic ring and a 5-7 membered aromatic heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O, and a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O;

R ₁ , R ₂ , R ₃ and R ₅ 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 ₆ R ₇ , S(O) ₂ R ₆ , S(O) ₂ NR ₆ R ₇ , NR ₆ R ₇ , C(O)NR ₆ R ₇ , C(O)NR ₆ S(O) ₂ R ₇ , NR ₆ S(O) ₂ R ₇ , C(O)R ₆ , NR ₆ C(O)R ₇ , a 5-12 membered heteroaryl having 1-3 heteroatoms selected from the following group consisting of N, S, and O, a 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the following 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 , R ₂ , R ₃ , and R ₅ may be substituted by 1-3 substituents independently selected from the following group: halogen, OH, CN, NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 haloalkoxy, NR ₆ R ₇ , C(＝O)NR ₆ R ₇ , C(＝O)NR ₆ S(＝O) ₂ R ₇ , P(＝O)R ₆ R ₇ , S(＝O) ₂ R ₆ , S(＝O) ₂ NR ₆ R ₇ , NR ₆ S(＝O) ₂ R ₇ ;

R ₄ is selected from the group consisting of H, deuterium, C(O)C _1-6 alkyl, C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, or C _3-12 heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S, and O, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclyl in the group represented by R ₄ 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, C _3-8 cycloalkyl, C _5-10 membered heterocyclyl, and NR ₆ R ₇ ;

R _a is selected from the following group: H, deuterium, C (O) C _1-6 alkyl, C _1-6 deuterated alkyl, C _1-6 alkyl, C _3-8 cycloalkyl, C _2-6 alkenyl, C _2-6 alkynyl or 3-12 membered heterocyclyl having 1-3 heteroatoms selected from the following group: N, S, O, wherein the alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl in the group represented by R _b may be substituted by 1-3 substituents independently selected from the following group: halogen, OH, CN, NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C 1-6 alkoxy, C _1-6 haloalkoxy, C _3-8 cycloalkyl, _5-10 membered heterocyclyl and NR ₆ R ₇ ;

R ₆ and R ₇ are each independently H or C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, C _3-12 heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S and O, wherein the alkyl, cycloalkyl, heterocyclyl represented by R ₆ or R ₇ 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 _3-8 cycloalkyl, C _3-12 heterocyclyl.

R ₆ and R ₇ together with the nitrogen atom or phosphorus atom to which they are attached may form a C _3-12 heterocyclic group, which may be substituted by 1-3 substituents independently selected from the following groups: 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;

n is 1, 2 or 3;

m is 1, 2 or 3; and n+m≤4;

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

t is 0, 1, or 2;

Furthermore, the compound of formula (I) has any structure shown below:

in:

X ₁ is selected from the group consisting of none, NR, O, S(O) _t , CHR, NRC(O), C(O)NR, NRC(O)C(O)NR, NRC(O)NR, NRC(S)NR, NRC(O)NRCH ₂ , NRC(S)NRCH ₂ , wherein R is selected from the group consisting of H, C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, or a 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O;

_X2 or _X3 are each independently selected from none, NR _a , O, S(O) _t , C(O), CR ₄ R ₅ ;

^M2 is CH or N; and when said ^M2 is CH, said May be located on said M ² (i.e. M ² is C);

The A ring is selected from the following group: H, C _3-8 cycloalkyl, 3-12 membered heterocyclyl, C _6-10 aromatic ring, 5-10 membered aromatic heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered aromatic ring and a 5-7 membered heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered aromatic ring and a 5-7 membered aromatic heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O, and a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O;

R ₁ , R ₂ , R ₃ and R ₅ 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 ₆ R ₇ , S(O) ₂ R ₆ , S(O) ₂ NR ₆ R ₇ , NR ₆ R ₇ , C(O)NR ₆ R ₇ , C(O)NR ₆ S(O) ₂ R ₇ , NR ₆ S(O) ₂ R ₇ , C(O)R ₆ , NR ₆ C(O)R ₇ , a 5-12 membered heteroaryl having 1-3 heteroatoms selected from the following group consisting of N, S, and O, a 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the following 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 , R ₂ , R ₃ , and R ₅ may be substituted by 1-3 substituents independently selected from the following group: halogen, OH, CN, NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 haloalkoxy, NR ₆ R ₇ , C(＝O)NR ₆ R ₇ , C(＝O)NR ₆ S(＝O) ₂ R ₇ , P(＝O)R ₆ R ₇ , S(＝O) ₂ R ₆ , S(＝O) ₂ NR ₆ R ₇ , NR ₆ S(＝O) ₂ R ₇ ;

R ₄ is selected from the group consisting of H, deuterium, C(O)C _1-6 alkyl, C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, or C _3-12 heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S, and O, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclyl in the group represented by R ₄ 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, C _3-8 cycloalkyl, C _5-10 membered heterocyclyl, and NR ₆ R ₇ ;

R _a is selected from the following group: H, deuterium, C (O) C _1-6 alkyl, C _1-6 deuterated alkyl, C _1-6 alkyl, C _3-8 cycloalkyl, C _2-6 alkenyl, C _2-6 alkynyl or 3-12 membered heterocyclyl having 1-3 heteroatoms selected from the following group: N, S, O, wherein the alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl in the group represented by R _b may be substituted by 1-3 substituents independently selected from the following group: halogen, OH, CN, NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C 1-6 alkoxy, C _1-6 haloalkoxy, C _3-8 cycloalkyl, _5-10 membered heterocyclyl and NR ₆ R ₇ ;

R ₆ and R ₇ are each independently H or C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, C _3-12 heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S and O, wherein the alkyl, cycloalkyl, heterocyclyl represented by R ₆ or R ₇ 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 _3-8 cycloalkyl, C _3-12 heterocyclyl.

R ₆ and R ₇ together with the nitrogen atom or phosphorus atom to which they are attached may form a C _3-12 heterocyclic group, which may be substituted by 1-3 substituents independently selected from the following groups: 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;

n is 1, 2 or 3;

m is 1, 2 or 3; and n+m≤4;

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

t is 0, 1, or 2;

Furthermore, the compound of formula (I) has any structure shown below:

In another aspect, the present invention provides a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt, isomer or hydrate thereof, the method comprising:

The compound of formula I is obtained by using a benzo 1,2,4-triazine compound to carry out Suzuki coupling, copper catalytic coupling, deprotection and other reactions.

In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of one or more of the above-mentioned compounds, or a pharmaceutically acceptable salt, isomer or hydrate thereof, and a pharmaceutically acceptable excipient.

In another aspect, the present invention provides a use of the above compound, or a pharmaceutically acceptable salt, isomer or hydrate thereof, in the preparation of a pharmaceutical composition for preventing or treating a disease responsive to inhibition of HPK1 activity in a subject.

Furthermore, the application includes a combination use of the above-mentioned compound or its pharmaceutically acceptable salt, isomer or hydrate with other tumor immunotherapeutic agents, and the other tumor immunosuppressants are selected from the following groups: small molecule compounds and antibodies (including but not limited to PD-1, PD-L1, CTLA-4, STING agonists, LAG3 antagonists, etc.), tumor targeted drugs, tumor vaccines, and radiotherapy regimens.

On the other hand, the present invention provides a use of the above-mentioned compound or its pharmaceutically acceptable salt, stereoisomer, prodrug, solvate, ester and deuterated compound in combination with CAR-T immunotherapy in cancer immunotherapy.

Furthermore, the disease is cancer.

In another aspect, the present invention provides a use of the above compound, or a pharmaceutically acceptable salt, isomer or hydrate thereof in an HPK1 kinase inhibitor.

Since the compounds of the present invention have very good HPK1 kinase inhibitory activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable organic or inorganic salts, solvates or hydrates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to prevent and/or treat diseases related to HPK1 kinase activity or expression (e.g., cancer).

The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable carrier or excipient. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

"Pharmaceutically acceptable carrier" refers to: one or more compatible liquid or solid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium ethyl cellulose, sodium carboxymethyl cellulose, 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.), colorants, polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween R, wetting agents (such as sodium lauryl sulfate), flavoring agents, preservatives, stabilizers, antioxidants, pyrogen-free water, etc.

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

Solid dosage forms for oral administration include capsules, pills, tablets, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as dicalcium phosphate or sodium citrate, or with the following ingredients: (a) fillers or extenders, for example, lactose, sucrose, starch, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, gelatin, alginates, polyvinyl pyrrolidone, gum arabic and sucrose; (c) humectants, for example, glycerol; (d) disintegrants, for example, calcium carbonate, agar, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; (i) lubricants, for example, talc, solid polyethylene glycols, calcium stearate, magnesium stearate, sodium lauryl sulfate or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, capsules, pills, granules and granules can be prepared using shell materials and coatings, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions may be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, suspensions, solutions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example: ethanol, isopropanol, propylene glycol, ethyl acetate, ethyl carbonate, 1,3-butylene glycol, dimethylformamide and oil, in particular peanut oil, cottonseed oil, corn germ oil, olive oil, sesame oil and castor oil or mixtures of these substances.

Besides such inert diluents, the composition may also include adjuvants such as emulsifying agents, wetting agents and suspending agents, flavoring agents, sweetening agents, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, polyoxyethylene sorbitol and sorbitan esters, ethoxylated isostearyl alcohol, microcrystalline cellulose, agar and aluminum methoxide, or mixtures of these substances and the like.

Compositions for parenteral injection may include physiologically acceptable sterile aqueous or dispersion solutions, anhydrous 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 suitable mixtures thereof.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When administered in combination, the pharmaceutical composition may further comprise one or more other pharmaceutically acceptable compounds. One or more other pharmaceutically acceptable compounds may be administered simultaneously, separately or sequentially with the compound of the present invention.

When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 20 to 500 mg. Of course, the specific dosage should also take into account factors such as the patient's health status and the route of administration, which are all within the skill of a skilled physician.

Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: it has excellent HPK1 inhibitory activity and can be used to prepare a pharmaceutical composition for treating cancer and other HPK activity-related diseases.

Detailed ways

The present invention provides a compound shown in the following formula I:

in:

X ₁ is selected from the group consisting of none, NR, O, S(O) _t , CHR, NRC(O), C(O)NR, NRC(O)C(O)NR, NRC(O)NR, NRC(S)NR, NRC(O)NRCH ₂ , NRC(S)NRCH ₂ , wherein R is selected from the group consisting of H, C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, or a 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the group consisting of N, S and O;

_X2 or _X3 are each independently selected from none, NR _a , O, S(O) _t , C(O), CR ₄ R ₅ ;

Y is selected from NR _a , O, S(O) _t or CR ₄ R ₅ ;

M ¹ , M ² , or M ³ are each independently CH or N; and when M ¹ , M ² , or M ³ is CH, may be located on the M ¹ , M ² , or M ³ (ie, M ¹ , M ² , or M ³ is C);

The A ring is selected from the following group: H, C _3-8 cycloalkyl, 3-12 membered heterocyclyl, C _6-10 aromatic ring, 5-10 membered aromatic heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered aromatic ring and a 5-7 membered heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered aromatic ring and a 5-7 membered aromatic heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heterocyclic ring having 1-3 heteroatoms selected from the following group N, S, O, a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O, and a 6-membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O and a 5-7 membered heteroaromatic ring having 1-3 heteroatoms selected from the following group N, S, O;

R ₁ , R ₂ , R ₃ and R ₅ 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 ₆ R ₇ , S(O) ₂ R ₆ , S(O) ₂ NR ₆ R ₇ , NR ₆ R ₇ , C(O)NR ₆ R ₇ , C(O)NR ₆ S(O) ₂ R ₇ , NR ₆ S(O) ₂ R ₇ , C(O)R ₆ , NR ₆ C(O)R ₇ , a 5-12 membered heteroaryl having 1-3 heteroatoms selected from the following group consisting of N, S, and O, a 3-12 membered heterocyclic group having 1-3 heteroatoms selected from the following 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 , R ₂ , R ₃ , and R ₅ may be substituted by 1-3 substituents independently selected from the following group: halogen, OH, CN, NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 haloalkoxy, NR ₆ R ₇ , C(＝O)NR ₆ R ₇ , C(＝O)NR ₆ S(＝O) ₂ R ₇ , P(＝O)R ₆ R ₇ , S(＝O) ₂ R ₆ , S(＝O) ₂ NR ₆ R ₇ , NR ₆ S(＝O) ₂ R ₇ ;

R ₄ is selected from the group consisting of H, deuterium, C(O)C _1-6 alkyl, C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, C _2-6 alkenyl, C _2-6 alkynyl, or C _3-12 heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S, and O, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, or heterocyclyl in the group represented by R ₄ 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, C _3-8 cycloalkyl, C _5-10 membered heterocyclyl, and NR ₆ R ₇ ;

R _a is selected from the following group: H, deuterium, C (O) C _1-6 alkyl, C _1-6 deuterated alkyl, C _1-6 alkyl, C _3-8 cycloalkyl, C _2-6 alkenyl, C _2-6 alkynyl or 3-12 membered heterocyclyl having 1-3 heteroatoms selected from the following group: N, S, O, wherein the alkyl, alkenyl, alkynyl, cycloalkyl or heterocyclyl in the group represented by R _b may be substituted by 1-3 substituents independently selected from the following group: halogen, OH, CN, NO ₂ , C _1-6 alkyl, C _1-6 haloalkyl, C 1-6 alkoxy, C _1-6 haloalkoxy, C _3-8 cycloalkyl, _5-10 membered heterocyclyl and NR ₆ R ₇ ;

R ₆ and R ₇ are each independently H or C _1-6 alkyl, C _1-6 deuterated alkyl, C _3-8 cycloalkyl, C _3-12 heterocyclyl having 1-3 heteroatoms selected from the group consisting of N, S and O, wherein the alkyl, cycloalkyl, heterocyclyl represented by R ₆ or R ₇ 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 _3-8 cycloalkyl, C _3-12 heterocyclyl.

R ₆ and R ₇ together with the nitrogen atom or phosphorus atom to which they are attached may form a C _3-12 heterocyclic group, which may be substituted by 1-3 substituents independently selected from the following groups: 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;

n is 1, 2 or 3;

m is 1, 2 or 3; and n+m≤4;

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

t is 0, 1, or 2;

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

Preferably, _X1 , _X2 , _X3 , Y, ^M1 , ^M2 , M3, M4, ^M5 , ^R , _R1 , ^R2 , _R3 , _R4 , _R5 , _R6 , _R7 , _{Ra ,} n, m, p, q, and t are each independently a corresponding group in the specific compounds in each embodiment.

The compounds of the embodiments of the present invention can be used as HPK1 kinase inhibitors, and in preferred embodiments, are HPK1 kinase selective inhibitors.

Preparation of compounds of formula I

The compound of formula I of the present invention can be prepared by the following exemplary method:

The compound of formula I is obtained by using a benzo 1,2,4-triazine 1,2,4-triazine compound to carry out Suzuki coupling, copper catalyzed coupling, deprotection and other reactions.

The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

Key intermediate 1. Preparation of tert-butyl (3-amino-5-bromobenzo[e][1,2,4]triazine-8-yl)carbamate

Step 1: At room temperature, slowly add Boc ₂ O (53.8 g, 247 mmol) to a THF solution (150 ml) of 4-bromo-2-nitroaniline (12.6 g, 57.95 mmol) and DMAP (10.6 g, 86.76 mmol), and heat to 70°C for 3 h. Cool to room temperature, concentrate under reduced pressure, and purify by column chromatography to obtain a light yellow compound (4-bromo-2-nitrophenyl)carbamic acid di-tert-butyl ester. MS (ESI): M/Z=418[M+H] ⁺ .

Step 2: At room temperature, a solution of potassium tert-butoxide (5.2 g, 46.4 mmol) in tetrahydrofuran (20 ml) was slowly added dropwise to a solution of (4-bromo-2-nitrophenyl) di-tert-butyl carbamate (2.42 g, 5.8 mmol) and guanidine hydrochloride (1.11 g, 11.6 mmol) in tetrahydrofuran (40 ml), and the temperature was raised to 65°C for overnight reaction. After the reaction was completed, the mixture was cooled to room temperature, saturated ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The target compound (3-amino-5-bromobenzo[e][1,2,4]triazine-8-yl) carbamate was purified by column chromatography to obtain a bright yellow target compound. MS (ESI): M/Z = 341 [M+H] ⁺ . Preparation of key intermediate 2.7-iodo-6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline

Step 1: At 0°C, add 7-methoxy-1,2,3,4-tetrahydroisoquinoline (6 g, 33.85 mmol) and concentrated nitric acid (3.46 g) in precooled trifluoroacetic acid (46 ml), stir at 0°C for 1 h, concentrate under reduced pressure, adjust pH to 10-11 with 4N NaOH aqueous solution, extract with DCM, dry the organic phase with anhydrous sodium sulfate, filter, spin dry, and purify by column chromatography to obtain 6-methoxy-7-nitro-1,2,3,4-tetrahydroisoquinoline (1.85 g, 24.6%). MS (ESI): M/Z=209[M+H] ⁺ .

Step 2: Add 37% formaldehyde solution (23 ml) to a solution of 7-methoxy-6-nitro-1,2,3,4-tetrahydroisoquinoline (7.58 g, 46.44 mmol) in methanol (82 mL) at 0°C, stir at 0°C for 15 minutes, then add NaBH ₄ (6.15 g) in batches, stir the resulting mixture at room temperature for 3 h, quench with ice water (20 mL), extract with DCM, dry the organic phase with anhydrous sodium sulfate, filter, spin dry, and purify by column chromatography to obtain 6-methoxy-2-methyl-7-nitro-1,2,3,4-tetrahydroisoquinoline. MS (ESI): M/Z=223[M+H] ⁺ .

Step 3: Add Fe powder (13.2 g, 237.60 mmol) and hydrochloric acid (1 ml) to a solution of water (10 ml) and EtOH (90 ml) containing 7-methoxy-2-methyl-6-nitro-1,2,3,4-tetrahydroisoquinoline (8.8 g, 39.6 mmol), stir at 60°C for 3 hours, cool to room temperature, filter, wash the filter cake with methanol, wash the filtrate with saturated sodium bicarbonate, concentrate the organic phase under reduced pressure, and purify by column chromatography to obtain 6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolin-7-amine. MS (ESI): M/Z = 193 [M+H] ⁺ .

Step 4: At room temperature, add p-toluenesulfonic acid monohydrate (1.71 g, 9 mmol) to a solution of 7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolin-6-amine (576.8 mg, 3 mmol) in acetonitrile (14 ml), cool to 0°C, slowly dropwise add KI (1.25 g, 7.5 mmmol) and NaNO ₂ (414 mg, 6 mmol) in water (6 ml), react at this temperature for 2 h and then slowly warm to room temperature. Add saturated sodium bicarbonate aqueous solution, extract with ethyl acetate, concentrate the organic phase under reduced pressure, and purify by column chromatography to obtain the target product 7-iodo-6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline. MS (ESI): M/Z=304[M+H] ⁺ .

Preparation of key intermediate 3.6-iodo-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline

Steps 1, 2, 3, and 4: The same operation process as that of key intermediate 2 was used to prepare 6-iodo-7-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinoline. MS (ESI): M/Z = 193 [M+H] ⁺ .

Preparation of key intermediate 4.8-iodo-9-methoxy-1,3,4,6,11,11a-hexahydro-2H-pyrido[1,2-b]isoquinoline

The steps are the same as the operation process of step 4 of key intermediate 2 to prepare 8-iodo-9-methoxy-1,3,4,6,11,11a-hexahydro-2H-pyrido[1,2-b]isoquinoline. MS (ESI): M/Z=344[M+H] ⁺ .

Preparation of key intermediate 5.6-fluoro-7-iodo-2-methyl-1,2,3,4-tetrahydroisoquinoline

Step 1: Potassium nitrate (578.47 mg, 5.72 mmol) was added in portions to a solution of 6-fluoro-3,4-dihydro-2H-isoquinolin-1-one (900 mg, 5.45 mmol) in H2SO4 (9 mL) at 0°C, reacted at 0°C for 2 h, poured into ice water, the precipitated solid was collected by filtration, and dried in vacuo to provide 6-fluoro-7-nitro-3,4-dihydro-2H-isoquinolin-1-one (1.04 g, 86%). MS (ESI): M/Z=221[M+H] ⁺ .

Step 2: Add 1M BH3/THF (45.46mmol, 45.5mL) to a THF (200mL) solution containing 6-fluoro-7-nitro-3,4-dihydro-2H-isoquinolin-1-one (2.02g, 9.09mmol), stir under reflux for 20 hours, carefully quench with methanol (30mL) under ice bath, concentrate the resulting solution in vacuo, add 2N HCl (50mL), heat at 80°C for 3 hours, add ammonia water to adjust the pH to 7-8, extract with DCM, dry the organic phase with anhydrous sodium sulfate, filter, and spin dry to obtain 6-fluoro-7-nitro-1,2,3,4-tetrahydroisoquinoline (1.89g, 100.00%). MS (ESI): M/Z=197[M+H] ⁺ .

Steps 3, 4, and 5: The same operation process as that of key intermediate 2 was used to prepare 6-fluoro-7-iodo-2-methyl-1,2,3,4-tetrahydroisoquinoline. MS (ESI): M/Z = 292 [M+H] ⁺ .

Key intermediate 6: Preparation of 6-methoxy-2-methylisoindol-5-amine

The operation process is the same as that of the key intermediate 5, except that 5-methoxyisoindol-1-one is used instead of 6-fluoro-3,4-dihydro-2H-isoquinolin-1-one. MS (ESI): M/Z=179 [M+H] ⁺ .

Preparation of key intermediate 7.8-methoxy-3-methyl-2,3,4,5-tetrahydro-1H-benzo[d]azepine-7-amine

The steps are the same as the operation process of key intermediate 6, and finally 8-methoxy-3-methyl-2,3,4,5-tetrahydro-1H-benzo[d]azepine-7-amine is prepared. MS (ESI): M/Z=207[M+H]+.

Preparation of key intermediate 8.3-iodo-2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthyridine

The steps are the same as the operation process of step 4 of key intermediate 2 to prepare 3-iodo-2-methoxy-6-methyl-5,6,7,8-tetrahydro-1,6-naphthyridine. MS (ESI): M/Z=305[M+H] ⁺ .

Preparation of key intermediate 9.5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinolin-1(2H)-one

5-Chloro-3,4-dihydroisoquinolin-1(2H)-one (3.4 g, 18.83 mmol), diboron dipinacol (23.93 g, 94.22 mmol), Pd(dppf)Cl2(II) (2.76 g, 3.77 mmol) and potassium acetate (5.55 g, 56.62 mmol) were added to 1,4-dioxane (2 mL) and stirred at 100° C. overnight under nitrogen protection. The solvent was concentrated under vacuum. The residue was purified by flash chromatography on silica gel to give 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinolin-1(2H)-one. MS (ESI): M/Z=274[M+H] ⁺ .

Preparation of key intermediate 10.5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester

The steps are the same as the operation process of key intermediate 9 to prepare tert-butyl 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-3,4-dihydroisoquinoline-2(1H)-carboxylate. MS (ESI): M/Z=360[M+H]+.

Example 1. 5-(2-bromophenyl)-N ³ -(6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)benzo[e][1,2,4]triazine-3,8-diamine

Step 1: Dissolve the key intermediate 1 (459 mg, 1.35 mmol) in a mixed solution of dioxane (21 ml) and water (3 ml), add 2-bromophenylboronic acid (542 mg, 2.7 mmol), Pd(dppf)Cl ₂ (197 mg, 0.27 mmol), sodium carbonate (429 mg, 4.05 mmol), reflux at 90°C for 3 h under nitrogen protection, cool to room temperature, filter with celite, concentrate the filtrate under reduced pressure, and purify by column chromatography to obtain tert-butyl (3-amino-5-(2-bromophenyl)benzo[e][1,2,4]triazine-8-yl)carbamate. MS (ESI): M/Z=417[M+H] ⁺ .

Step 2: Dissolve the (3-amino-5-(2-bromophenyl)benzo[e][1,2,4]triazine-8-yl)carbamic acid tert-butyl ester (291.4 mg, 0.7 mmol) obtained in the previous step in a mixed solution of toluene (4 ml) and tetrahydrofuran (0.8 ml), add key intermediate 2 (212.1 mmol, 0.7 mmol), cuprous iodide (136.5 mg, 0.7 mmol), N,N'-dimethylethylenediamine (61.6 mg, 0.7 mmol), potassium phosphate (446 mg, 2.1 mmol), reflux at 110 ° C for 24 h under nitrogen protection, cool to room temperature, filter with diatomaceous earth, dry the filtrate under reduced pressure, and purify by column chromatography to obtain 5-(2-bromophenyl)-N ³ -(6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)benzo[e][1,2,4]triazine-3,8-diamine. MS (ESI): M/Z=592[M+H] ⁺ .

Step 3: To a solution of tert-butyl (5-(2-bromophenyl)-3-((6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)amino)benzo[e][1,2,4]triazine-8-yl)carboxylate (59.1 mg, 0.1 mmol) in dichloromethane (5 ml), trifluoroacetic acid (1 ml) was added and stirred at room temperature for 2 h. Saturated sodium bicarbonate was added to adjust the pH to neutral, and the mixture was extracted with dichloromethane three times. The organic phases were combined, dried under reduced pressure, and purified by column chromatography to obtain the orange-red target compound 5-(2-bromophenyl)-N ³ -(6-methoxy-2-methyl-1,2,3,4-tetrahydroisoquinolin-7-yl)benzo[e][1,2,4]triazine-3,8-diamine. MS (ESI): M/Z=492[M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δ8.56 (s, 1H), 7.84 (dd, J = 1.4, 7.7 Hz, 1H), 7.75 ( d, J = 8.3 Hz, 1H), 7.66 ( dd, J = 1.2, 7.8 Hz, 1H), 7.51 ( td, J = 1.3, 7.6 Hz, 1H), 7.41–7 .31(m,2H),7.01(d,J＝8.5 Hz,1H),6.61(t,J＝0.9 Hz,1H),5.71(s,2H),3.86(s,3H),3.68(dd,J＝1.0,13.6 Hz,1H),3.36(dd,J＝1.0,13.6Hz,1H),3.00–2.86(m,3H),2.75–2.63(m,1H),2.43(s,3H).

Bioactivity test example 1HPK1 ADP-Glo enzyme activity test

Prepare enzyme activity test buffer containing 40mM Tris, pH7.5; 20mM MgC12; 0.1mg/ml BSA; 50uM DTT. Dissolve the compound in pure DMSO, and the stock concentration is 10mM. The compound DMSO solution is diluted three times from 100uM, for a total of 11 concentrations. The diluted compound is diluted 1:20 with enzyme activity test buffer, and 1ul is added to the working well, with two replicates for each concentration. Add 1ul of DMSO solution diluted 1:20 to both the negative control well and the positive control well. Prepare 2.5× substrate/ATP working solution as enzyme activity test buffer containing 45uM ATP and 0.25ug/ul MBP protein, and add 2ul of 2.5× substrate/ATP working solution to each working well. Prepare 2.5× enzyme reaction working solution as enzyme activity test buffer containing 0.5ng/ul HPK1 recombinant protein (Signalchem, catalog number M23-11G-10), add 2ul of 2.5× enzyme reaction working solution to each working well, and add only 2ul of enzyme activity test buffer to the negative control well. Stick the sealing film and centrifuge, and place it at room temperature for 30 minutes. After the reaction, add 5ul of ADP-Glo reagent (Promega, catalog number V1901) to each well and continue to react at room temperature for 40 minutes. Add 10ul of kinase detection reagent (Promega, catalog number V1901) and react at room temperature for 20 minutes before measuring the final luminescent signal.

The average values of the positive wells and negative wells were calculated as the positive control value (Signalpos) and the negative control value (Signalneg). The inhibition rate of the working well signal value (Signaltest) was calculated according to the formula Inhibition rate = (Signalpos-Signaltest)/(Signalpos-Signalneg) × 100%. The calculated inhibition rate was used to draw a concentration-inhibition rate curve in GraphPadPrism software by nonlinear fitting, and the IC50 value was calculated.

Biological Activity Test Example 2 GLK ADP-Glo Enzyme Activity Test

Prepare enzyme activity test buffer containing 40mM Tris, pH7.5; 20mM MgC12; 0.1mg/ml BSA; 50uM DTT. Dissolve the compound in pure DMSO, and the stock concentration is 10mM. The compound DMSO solution is diluted three times from 100uM, for a total of 11 concentrations. The diluted compound is diluted 1:20 with enzyme activity test buffer, and 1ul is added to the working well, with two replicates for each concentration. Add 1ul of DMSO solution diluted 1:20 to both the negative control well and the positive control well. Prepare 2.5× substrate/ATP working solution as enzyme activity test buffer containing 105uM ATP and 0.5ug/ul PKA substrate peptide, and add 2ul of 2.5× substrate/ATP working solution to each working well. Prepare 2.5× enzyme reaction working solution containing 2.5ng/ul GLK recombinant protein (Signalchem, Catalog No. M25-11G-10) enzyme activity test buffer, add 2ul of 2.5× enzyme reaction working solution to each working well, and add only 2ul of enzyme activity test buffer to the negative control well. Stick the sealing film and centrifuge, and place it at room temperature for 1 hour. After the reaction, add 5ul of ADP-Glo reagent (Promega, Catalog No. V1901) to each well and react at room temperature for 40 minutes, then add 10ul of kinase detection reagent (Promega, Catalog No. V1901) and react at room temperature for 20 minutes, and measure the final luminescent signal.

The average values of the positive wells and negative wells were calculated as the positive control value (Signalpos) and the negative control value (Signalneg). The inhibition rate of the working well signal value (Signaltest) was calculated according to the formula Inhibition rate = (Signalpos-Signaltest)/(Signalpos-Signalneg) × 100%. The calculated inhibition rate was used to draw the concentration-inhibition rate curve in the GraphPadPrism software according to nonlinear fitting, and the IC50 value was calculated. The experimental results are shown in the following table:

Table 1 Results of HPK1 and GLK enzyme activity test

Among them, A means IC ₅₀ value ≤ 50nM; B means 50nM ≤ IC ₅₀ value ≤ 500nM; C means 500nM ≤ IC ₅₀ value ≤ 10uM.

Claims (7)

1. A compound represented by the following formula (I) or a pharmaceutically acceptable salt thereof: in: _X2 and _X3 are _{CR4R5 , R4} and _R5 are H; ^M2 is CH; Ring A is selected from the following group: a C _6-10 aromatic ring, a 6-membered aromatic ring and a 5-7-membered heterocyclic ring having 1-3 heteroatoms selected from the following group: N, S, O; _R1 is _{NR6R7 , R6} and _R7 are H; R ₂ is selected from C _1-6 alkyl substituted by C _1-6 haloalkyl, OH or C _1-6 alkoxy, S(O) ₂ NR ₆ R ₇ , R ₆ and R ₇ are H; R ₃ is halogen or C _1-6 alkoxy; Ra is a C _1-6 alkyl group; q is 0, 1, 2, 3, 4 or 5. 2. A compound or a pharmaceutically acceptable salt thereof, characterized in that it has any structure shown in the following formula: 3. A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds as claimed in claim 1 or 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. 4. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of an HPK1 kinase inhibitor. 5. Use of the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in the preparation of an anti-tumor drug, wherein the tumor is a tumor associated with HPK1 kinase. 6. The use according to claim 4, characterized in that the use comprises a combination use of the compound or a pharmaceutically acceptable salt thereof with other tumor immunotherapeutic agents, wherein the other tumor immunosuppressants are selected from the following groups: small molecule compounds and antibodies, tumor targeted drugs, tumor vaccines, and radiotherapy regimens. 7. Use of a compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof in combination with CAR-T immunotherapy in the preparation of cancer immunotherapy drugs.