WO2021068380A1 - Preparation of degradation compound targeting btk protein, and application thereof in treating autoimmune diseases and tumors - Google Patents

Abstract

A compound, the compound being a compound represented by formula I, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof. The compound has a strong degradation effect on wild-type BTK, has no inhibition or degradation effects on other targets such as EGFR, ITK and TEC, and has an effect of specifically targeting and degrading BTK proteins. X-Y-Z formula I

Description

Preparation of a compound targeting BTK protein degradation and its application in the treatment of autoimmune system diseases and tumors Technical field

The present invention relates to the field of biomedicine. Specifically, the present invention relates to the preparation of a compound targeting BTK protein degradation and its application in the treatment of autoimmune system diseases and tumors.

Background technique

PROTAC (Proteolysis Targeting Chimeras) technology, or protein targeted degradation technology, is a new chemical biology or drug discovery method that uses the ubiquitin-proteasome system to induce protein degradation in recent years. The role of PROTAC small molecules is based on the process of E3 ubiquitin ligase ligating a certain protein. One end of the PROTAC small molecule chimera can specifically bind to the target protein, and the other end recruits E3 ubiquitin ligase. Bring it close to the target protein, and then the subsequent degradation process occurs. The PROTAC chimera is synthesized on the basis of selective small molecule inhibitors. Through clever design, it can distinguish highly homologous proteins. This gives PROTAC technology a certain advantage in the application of biological tools; in addition, PROTAC molecules and The target proteins are non-covalently bonded. After the degradation of one molecule of protein, it can be dissociated to degrade another molecule of target protein. This catalytic amount of protein degradation is the development of high-activity and low-toxic drug molecules. It provides the possibility that PROTAC technology has received widespread attention and application because it is more direct and fast.

BTK is a member of the non-receptor tyrosine kinase family and a key kinase in the B cell antigen receptor (BCR) signaling pathway. It can control the development and differentiation of B cells by activating positive cell cycle regulators and differentiation factors. It can regulate the survival and proliferation of B cells through the expression of pro-apoptotic and anti-apoptotic proteins. The continuous activation of BTK is a prerequisite for the development of many hematological diseases and autoimmune diseases. Therefore, the regulation of BTK protein has a good prospect for the treatment of hematological malignancies and autoimmune disorders.

Non-Hodgkin's lymphoma (NHL) is a blood system cancer, which is the general term for all lymphomas except Hodgkin's lymphoma. In the United States, 2.1% of people’s lives are affected by it. In 2015, 4.3 million people suffered from non-Hodgkin's lymphoma, and 231400 were killed. Most clinical non-Hodgkin's lymphomas are B-cell type, accounting for 70%-85% of the total. Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin's lymphoma (NHL). In the United States and the United Kingdom, 7-8 people out of 100,000 people are affected each year. Ibrutinib has been approved by the FDA for the treatment of chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL) and Waldenstrom's macroglobulinemia (WM) indications. And there are reports that ibrutinib can effectively inhibit the proliferation of some types of DLBCL. Ibrutinib works by covalently cross-linking its acrylamide structure with the sulfhydryl group of the cysteine 481 of the BTK protein in the cell, so that BTK loses its ability to phosphorylate downstream signaling proteins, thereby exerting anti-cell proliferation. The role of.

Although ibrutinib shows strong inhibitory activity against BTK kinase, the drug itself can cause many side effects due to its strong off-target effect. The IC ₅₀ value for EGFR is 5.6 nM, which can cause severe diarrhea and rash; the IC ₅₀ value for ITK is 10.7 nM, which can cause loss of natural killer cell function; the IC ₅₀ value for TEC is 78 nM, which can cause coagulation defects, and As a technology that can accurately and rapidly degrade target proteins, PROTAC has great advantages in realizing highly effective anti-tumor and reducing off-target side effects.

Autoimmune diseases refer to diseases caused by the body's immune response to self-antigens and damage to its own tissues, including pulmonary hemorrhagic nephritis syndrome, pemphigus and other organ-specific autoimmune diseases, arthritis, systemic lupus erythematosus and other systems The specific mechanism of sexual autoimmune diseases is not yet fully clarified, and one of the possible mechanisms is that the excessive expression of BTK activates the BCR signaling pathway abnormally, which in turn causes B cell dysfunction, changes in immune tolerance, and transforms into autoreactive B Cells secrete large amounts of autoantibodies to induce disease.

Based on the above mechanism, there have been more and more studies on the application of BTK inhibitors to autoimmune diseases (including rheumatoid arthritis) in recent years. Although only Ibrutinib was approved by the FDA in 2017 for autoimmune diseases (chronic graft-versus-host disease (cGVHD)), more than a dozen small molecule inhibitors have entered the clinical stage. BTK has become the most common target for clinical drugs in the field of autoimmune diseases except TNF (tumor necrosis factor) and CD20. It is expected to become a new target for the treatment of autoimmune diseases in the future. This is also the PROTAC technology that can efficiently degrade BTK protein. Provides opportunities and challenges.

Therefore, BTK kinase degradation agents have good prospects as drugs for anti-tumor or treatment of autoimmune diseases and need further development.

Summary of the invention

This application is based on the inventor's discovery and understanding of the following facts and problems:

In the prior art, Ibrutinib can inhibit BTK kinase, but the literature (N.Engl.J.Med.370, 2352-2254 (2014).) reported that Ibrutinib's BTK inhibitory ability against C481S mutation IC ₅₀ = 1 μM, and the drug is more effective than others. Targets, such as EGFR, ITK, and TEC also have varying degrees of effect, with obvious side effects. Based on the findings of the above-mentioned problems, the inventors proposed a new compound that uses a dual-target molecular structure. The structure is shown in Figure 1. One end of this type of molecule is targeted to bind E3 ligase, and the other end is a target. To bind to the target protein (BTK protein) to be degraded, the structures at these two ends are connected by a linker to form a complete compound molecule. The compound has a strong effect on the degradation of wild-type BTK, and has no inhibitory or degradation effect on other targets such as EGFR, ITK, TEC, etc., and has the effect of specifically targeting BTK protein.

For this reason, in the first aspect of the present invention, the present invention proposes a compound, which is a compound represented by formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates , Solvates, metabolites, pharmaceutically acceptable salts or prodrugs:

X-Y-Z

Formula I

among them:

The Y is

且各E独立地为酰胺基，酯基，氨基甲酯基，脲基，胍基，杂环基，环烷基，螺杂双环基，稠合杂双环基，桥杂双环基或C6-10芳基；各E任选地被1，2，3或4个独立的R _b所取代；各W为O、S、NH或Se，b为1～30之间的整数，d为0～30之间的整数，L ₆、L ₇独立为键，-O-，-S(＝O) _t1-，-S-，-N(R _a)-，-C(＝O)O-，-N(R _a)-C(＝O)-，-C(＝O)-(CH ₂) _t2-，-CH ₂-，-C(＝O)-，-OC(＝O)-，-C(＝S)-，-C(＝O)-N(R _a)-，-C(＝S)-N(R _a)-，-(CH ₂) _t2-C(＝O)-或者三氮唑基，

And each E is independently an amide group, an ester group, a carbamate group, a ureido group, a guanidyl group, a heterocyclic group, a cycloalkyl group, a spiro heterobicyclic group, a fused heterobicyclic group, a bridged heterobicyclic group or C6-10 Aryl; each E is optionally substituted by 1, 2, 3 or 4 independent R _b ; each W is O, S, NH or Se, b is an integer between 1-30, and d is 0-30 integer, L _6, L ₇ is independently a bond, -O -, - S (= O) t1 -, - S -, - N (R a) -, - C (= O) O -, - N (R _a )-C(=O)-, -C(=O)-(CH ₂ ) _t2 -, -CH ₂ -, -C(=O)-, -OC(=O)-, -C( =S)-, -C(=O)-N(R _a )-, -C(=S)-N(R _a )-, -(CH ₂ ) _t2 -C(=O)- or triazole base,

The X is

The Z is:

Or the Y is

d为0～30之间的整数，其中，

d is an integer between 0 and 30, where,

所述Z为：

d=6, L ₆ and L _{7 are} independently -N(R _a )-, when -C(=O)-, the X is

The Z is:

所述Z为：

d≠6, L ₆ and L _{7 are} independently bonds, -O-, -S(=O) _t1 -, -S-, -N(R _a )-, -C(=O)O-, -N( R _a )-C(=O)-, -CH ₂ -, -C(=O)-, -OC(=O)-, -C(=S)-, -C(=O)-N(R _a )-,-C(=S)-N(R _a )- or triazolyl, the X is

The Z is:

Or the Y is

且W为CH ₂、O、S、NH或Se，b为0～30之间的整数，d为0～30之间的整数，

And W is CH ₂ , O, S, NH or Se, b is an integer between 0 and 30, and d is an integer between 0 and 30,

The X is

The Z is:

Or the Y is

且W为CH ₂、O、S、NH或Se，b为0～30之间的整数，d为0～30之间的整数，

And W is CH ₂ , O, S, NH or Se, b is an integer between 0 and 30, and d is an integer between 0 and 30,

The X is

时，所述Z为：

When, the Z is:

The X is

时，所述Z为

时，The Z is

Wherein, each of L ₁ , L ₂ , and L _{3 is} independently a bond, -O-, -S(=O) _t1 -, -S-, -N(R _a )-, -C(=O)O-,- N(R _a )-C(=O)-, -C(=O)-(CH ₂ ) _t2 -, -CH ₂ -, -C(=O)-, -OC(=O)-, -C (=S)-, -C(=O)-N(R _a )-, -C(=S)-N(R _a )-, -(CH ₂ ) _t2 -C(=O)- or trinitrogen Azole

Each R _a is independently hydrogen, C1-4 alkyl, halo C1-4 alkyl, C1-4 alkyl group or a hydroxyl group;

Each t2 is independently 0, 1, 2, 3 or 4;

Each t1 is independently 0, 1 or 2;

Each of X ₁ , X ₂ , X ₃ , X ₄ , X ₅ , X ₆ , X ₇ , X ₈ , and Q is independently CH, CH ₂ , O, S, N, NH or Se;

Each R ₁ , R ₂ , R ₃ , R ₄ , R _{5 is} independently H, deuterium, amino, C1-4 amide, C1-4 alkyl, C1-4 heteroalkyl, C3-8 cycloalkyl, C2-10 heterocyclyl, C6-10 aryl, C1-9 heteroaryl, C6-10 aryl, C1-4 alkoxy, C1-4 alkenyl, C1-4 alkynyl, wherein, the C1-4 alkyl, C1-4 heteroalkyl, C3-8 cycloalkyl, C2-10 heterocyclyl, C6-10 aryl, C1-9 heteroaryl, C6-10 aryl, C1-4 alkane The oxy, C1-4 alkenyl, and C1-4 alkynyl groups can be optionally selected by one or more selected from deuterium, hydroxyl, amino, oxo, F, Cl, Br, I, cyano, C1-6 alkyl , C1-6 haloalkyl, C1-6 hydroxyalkyl, C1-6 aminoalkyl, C1-6 alkoxy C1-6 alkyl, C1-6 alkylamino C1-6 alkyl, C6-10 aryl C1 -6 alkyl, C1-9 heteroaryl C1-6 alkyl, C2-10 heterocyclyl C1-6 alkyl, C3-10 cycloalkyl C1-6 alkyl, C1-6 alkoxy, C1- 6Alkylamino, C6-10 aryl, C6-10 aryloxy, C6-10 arylamino, C1-9 heteroaryl, C1-9 heteroaryloxy, C2-6 alkenyl, C3-10 cycloalkyl , Halogen-substituted C6-10 aryl, halogen-substituted C1-9 heteroaryl, halogen-substituted C2-10 heterocyclic group or C2-10 heterocyclic group substituted;

Each A is independently hydrogen, C1-4 alkyl, C1-4 haloalkyl, hydroxy, nitro, amino, cyano, halogen, carboxy, C1-4 alkoxy, C1-4 alkylamino, C1-4 alkylthio Group, C1-4 alkyl acyl group, C3-12 cycloalkyl group, optionally substituted C3-9 heterocyclic group, optionally substituted C6-12 aryl group, optionally substituted C1-9 heteroaryl group;

Each R _{b is} independently hydrogen, C1-4 alkyl, C1-4 haloalkyl, hydroxy, nitro, amino, cyano, halogen, carboxy, C1-4 alkoxy, C1-4 alkylamino, C1-4 Alkylthio, C1-4 alkyl acyl, C3-12 cycloalkyl, C3-9 heterocyclyl, C6-12 aryl, C1-9 heteroaryl, amino C1-4 alkyl, hydroxy C1-4 alkane Group, sulfonic acid group, aminosulfonyl group or aminoacyl group;

Each of a, c, e, and f is independently an integer between 0-30.

According to an embodiment of the present invention, the above-mentioned compound may further include at least one of the following additional technical features:

According to an embodiment of the present invention, each L _{1 is} independently a bond, -O-, -S-, -NH-,

Each L _{2 is} independently a bond, -(C=O)CH=CH ₂ -, -O-, -S-, -S(=O)-, -S(=O) ₂ -, -NRa-,- C(=O)-, -C(=O)O-, -N(R _a )-C(=O)-, -CH ₂ -, -OC(=O)-, -C(=S)- , -C(=O)-N(R _a )-, -C(=S)-N(R _a ) or triazolyl,

Each R _a is independently hydrogen, C1-2 alkyl, halo C1-2 alkyl, C1-2 alkyl group or a hydroxyl group,

Each R ₁ , R ₂ , R ₃ , R _{4 is} independently H, amino, C1-4 alkyl, C1-4 heteroalkyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aromatic Group, C5-7 heteroaryl, wherein the C1-4 alkyl, C1-4 heteroalkyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aryl, C5-7 hetero The aryl group may be optionally substituted with one or more substituents selected from deuterium, hydroxyl, amino, oxo, F, Cl, Br, I, cyano.

According to an embodiment of the present invention, each X is independently a compound as shown below:

According to an embodiment of the present invention, each A is independently hydrogen, optionally substituted C5-7 heterocyclyl, C5-7 cycloalkyl, optionally substituted C6-7 aryl, optionally substituted C5-7 heteroaryl base,

Each R _{5 is} independently H, amino, C1-4 alkyl, C1-4 heteroalkyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aryl, C5-7 heteroaryl, Wherein, the C1-4 alkyl group, C1-4 heteroalkyl group, C5-7 cycloalkyl group, C5-7 heterocyclic group, C6-7 aryl group, C5-7 heteroaryl group may be optionally substituted by one or Multiple selected from deuterium, hydroxyl, amino, oxo, F, Cl, Br, I, cyano, C5-7 aryl, C5-7 heteroaryl, halogen substituted C5-7 aryl, halogen substituted C5-7 Heteroaryl, halogen-substituted C5-7 heterocyclyl substituents;

Each L _{3 is} independently a bond, -(C=O)CH=CH ₂ -, -O-, -S-, -S(=O)-, -S(=O) ₂ -, -NRa-,- C(=O)-, -C(=O)O-, -N(R _a )-C(=O)-, -CH ₂ -, -OC(=O)-, -C(=S)- , -C(=O)-N(R _a )-, -C(=S)-N(R _a ) or triazolyl,

Each R _a is independently hydrogen, C1-2 alkyl, halo C1-2 alkyl, C1-2 alkyl group or a hydroxyl group.

各R _c独立地为氢或C1-2烷基。 According to the embodiment of the present invention, each A is independently

Each R _{c is} independently hydrogen or C1-2 alkyl.

According to an embodiment of the present invention, each Z is independently a compound as shown below:

According to embodiments of the present invention, each E is independently an amide group, an ester group, a carbamate group, a ureido group, a guanidino group, a heterocyclic group, a cycloalkyl group or a C6-C8 aryl group; each E is optionally 1, Replaced by 2, 3 or 4 independent R _b,

Each R _{b is} independently hydrogen, C1-2 alkyl, C1-2 haloalkyl, hydroxy, nitro, amino, cyano, halogen, carboxy, C1-2 alkoxy, C1-2 alkylamino, C1-2 Alkylthio, C1-2 alkyl acyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aryl, C5-7 heteroaryl, amino C1-2 alkyl, hydroxy C1-2 alkane Group, sulfonic acid group, aminosulfonyl group or aminoacyl group,

Each R _a is independently hydrogen, C1-2 alkyl, halo C1-2 alkyl, C1-2 alkyl group or a hydroxyl group.

According to the embodiment of the present invention, each Y is independently

According to an embodiment of the present invention, the X is

In some embodiments, the Z is

In some embodiments, the Y is

Wherein, m1 is an integer between 1-10, and n1 is an integer between 0-10.

According to an embodiment of the present invention, the compound includes the compound represented by any one of formulas (1) to (15) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, Solvates, metabolites, pharmaceutically acceptable salts or prodrugs,

Wherein, m1 is an integer between 1-10, and n1 is an integer between 0-10.

In the second aspect of the present invention, the present invention proposes a compound. According to an embodiment of the present invention, it is a compound represented by any one of formulas 1-18 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, Pharmaceutically acceptable salts or prodrugs,

The compound according to the embodiment of the present invention has a strong effect on the degradation of wild-type BTK, and has no inhibitory or degradation effect on other targets such as EGFR, ITK, TEC, etc., and has the effect of specifically targeting and degrading BTK protein.

In the third aspect of the present invention, the present invention provides a pharmaceutical composition comprising the compound described above. The pharmaceutical composition according to the embodiment of the present invention has a strong effect on the degradation of wild-type BTK, and has no inhibitory or degradation effect on other targets such as EGFR, ITK, TEC, etc., and has the effect of specifically targeting and degrading BTK protein.

According to an embodiment of the present invention, the above-mentioned pharmaceutical composition may further include at least one of the following additional technical features:

According to an embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or a combination thereof.

According to an embodiment of the present invention, the pharmaceutical composition further includes other drugs for treating or preventing non-Hodgkin's lymphoma or autoimmune diseases. The inventor found that the combination of drugs has better effects in the treatment or prevention of non-Hodgkin's lymphoma or autoimmune diseases.

According to an embodiment of the present invention, the autoimmune disease is arthritis, pulmonary hemorrhage, systemic lupus erythematosus, corpus sores, chronic lymphatic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, chronic ulcerative disease Colitis, pernicious anemia with chronic atrophic gastritis, primary biliary cirrhosis, multiple cerebrospinal sclerosis or acute idiopathic polyneuritis. In some embodiments, the autoimmune disease is arthritis or pulmonary hemorrhage.

According to an embodiment of the present invention, the other drugs for treating or preventing non-Hodgkin's lymphoma or autoimmune diseases include ibrutinib.

In the fourth aspect of the present invention, the present invention proposes the use of the aforementioned compound or the aforementioned pharmaceutical composition in the preparation of a medicine, which is used to degrade BTK or inhibit BTK. The compound or pharmaceutical composition according to the embodiment of the present invention has a strong effect on the degradation of wild-type BTK, and has no inhibitory or degradation effect on other targets such as EGFR, ITK, TEC, etc., and has the effect of specifically targeting the BTK protein.

In the fifth aspect of the present invention, the present invention proposes the use of the aforementioned compound or the aforementioned pharmaceutical composition in the preparation of a medicine for the treatment or prevention of BTK-related diseases. The compound or pharmaceutical composition according to the embodiment of the present invention has a strong effect on the degradation of wild-type BTK, and has no inhibitory or degradation effect on other targets such as EGFR, ITK, TEC, etc., and has the effect of specifically targeting the BTK protein. Related diseases have good therapeutic or preventive effects.

According to the embodiment of the present invention, the above-mentioned use may further include at least one of the following additional technical features:

According to an embodiment of the present invention, the BTK-related disease is non-Hodgkin's lymphoma or an autoimmune disease. The compounds or pharmaceutical compositions according to the embodiments of the present invention have better treatment or prevention effects on non-Hodgkin's lymphoma or autoimmune diseases.

According to an embodiment of the present invention, the autoimmune disease is arthritis, pulmonary hemorrhage, systemic lupus erythematosus, corpus sores, chronic lymphatic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, chronic ulcerative disease Colitis, pernicious anemia with chronic atrophic gastritis, primary biliary cirrhosis, multiple cerebrospinal sclerosis or acute idiopathic polyneuritis.

According to an embodiment of the present invention, the autoimmune disease is arthritis or pulmonary hemorrhage. The inventor found that the compound or pharmaceutical composition described above can prevent and treat the symptoms of arthritis and pulmonary hemorrhage in a mouse model, and the prevention and improvement of the situation are significantly better than ibrutinib.

In another aspect of the present invention, the present invention proposes a general procedure for the synthesis of the compound represented by formula I. According to an embodiment of the present invention, the compound represented by formula I can be connected by click reaction or amide condensation reaction between Pomalidomide or Lenanidomide or RG-7112 terminal derivative and Ibrutinib terminal derivative, as shown in Figure 2, where, For the preparation method of Pomalidomide terminal derivatives, please refer to Literature & Biology 22,755-763 (2015). For the preparation method of Lenalidomide terminal derivatives, please refer to the literature J. Med.Chem (DOI: 10.1021/acs.jmedchem.6b01816)., RG-7112 For the preparation method of the terminal derivative, please refer to the literature Bioorg.Med.Chem.Lett.18,5904-5908 (2008).ACS Med.Chem.Lett.4,466-469(2013). The synthetic route of the terminal derivative of Ibrutinib can be referred to Patent PCT Int.Appl.,2013003629,03Jan 2013. The terminal alkynes required for Click Chemistry are connected to the piperidine ring of the Ibrutinib core through an amide condensation reaction. The preparation method can refer to the document J.Chem.Inf.Model.50,446(2010).PCT Int.Appl.,2013170115, 14 Nov 2013.

Description of the drawings

Figure 1 is a basic technical route of PROTACs according to an embodiment of the present invention;

Fig. 2 is a schematic diagram showing the construction of Formula I through a click reaction and an amide condensation reaction according to an embodiment of the present invention;

Fig. 3 is the inhibitory result of the compound according to an embodiment of the present invention on the release of inflammatory factors in THP-1 cells stimulated by LPS;

Fig. 4 shows the degradation effect of compounds according to examples of the present invention on BTK.

Detailed ways

The embodiments of the present invention are described in detail below, and examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the drawings are exemplary, and are intended to explain the present invention, but should not be construed as limiting the present invention.

As used herein, the term "administer to the patient the aforementioned compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable The salt or prodrug or the aforementioned pharmaceutical composition "refers to the introduction of a predetermined amount of a substance into a patient in a suitable manner. The compound of formula I of the present invention or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs or drugs thereof The composition can be administered by any common route, as long as it can reach the desired tissue. Various modes of administration are contemplated, including peritoneal, intravenous, intramuscular, subcutaneous, cortical, oral, topical, nasal, pulmonary, and rectal, but the present invention is not limited to these exemplified modes of administration. However, due to oral administration, the active ingredient of the oral administration composition should be coated or formulated to prevent it from being degraded in the stomach. In addition, the compound of formula I or the pharmaceutical composition of the present invention can be administered using a specific device that delivers the active ingredient to the target cell.

The administration frequency and dosage of the pharmaceutical composition of the present invention can be determined by a number of related factors, including the type of disease to be treated, the route of administration, the patient’s age, sex, body weight, and the severity of the disease, as well as the active ingredient Type of drug.

The term "therapeutically effective amount" refers to an amount of a compound that is sufficient to significantly improve certain symptoms associated with a disease or disorder, that is, an amount that provides a therapeutic effect for a given disorder and dosage regimen. A therapeutically effective amount of the drug or compound does not need to cure the disease or condition, but will provide treatment for the disease or condition so that the onset of the disease or condition of the individual is delayed, prevented, or prevented, or the symptoms of the disease or condition are alleviated, or the disease or condition The duration of the illness is changed, or, for example, the disease or illness becomes less serious, or recovery is accelerated.

The term "treatment" is used to refer to obtaining the desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of completely or partially preventing the disease or its symptoms, and/or may be therapeutic in terms of partially or completely curing the disease and/or adverse effects caused by the disease. "Treatment" as used herein encompasses the treatment of diseases in mammals, especially humans, including: (a) preventing the occurrence of diseases or disorders in individuals who are susceptible to but have not yet been diagnosed with the disease; (b) inhibiting the disease; or (c) Alleviate the disease, such as alleviating the symptoms associated with the disease. "Treatment" as used herein encompasses any medication that administers a drug or compound to an individual to treat, cure, alleviate, ameliorate, alleviate, or inhibit the individual’s disease, including but not limited to combining compounds or drugs containing formula I or formula II as described herein The giving of things to individuals in need.

According to an embodiment of the present invention, the excipients include pharmaceutically acceptable excipients, lubricants, fillers, diluents, disintegrants, stabilizers, preservatives, emulsifiers, cosolvents, and coloring agents that are well-known in the formulation field. , Sweetener, made into tablets, pills, capsules, injections and other different dosage forms.

Now certain embodiments of the present invention will be described in detail, examples of which are illustrated by the accompanying structural formulas and chemical formulas. The present invention intends to cover all alternatives, modifications and equivalent technical solutions, which are all included in the scope of the present invention as defined by the claims. Those skilled in the art should recognize that many methods and materials similar or equivalent to those described herein can be used to practice the present invention. The invention is by no means limited to the methods and materials described herein. In the case where one or more of the combined documents, patents and similar materials are different from or contradictory to this application (including but not limited to the defined terms, term application, described technology, etc.), this Application shall prevail.

It should be further recognized that certain features of the present invention, for the sake of clarity, have been described in multiple separate embodiments, but may also be provided in combination in a single embodiment. Conversely, the various features of the present invention are described in a single embodiment for the sake of brevity, but they can also be provided individually or in any suitable sub-combination.

Unless otherwise specified, all scientific and technological terms used in the present invention have the same meanings as commonly understood by those skilled in the art to which the present invention belongs. All patents and publications related to the present invention are incorporated into the present invention in its entirety by reference.

Unless otherwise stated, the following definitions used herein should be applied. For the purpose of the present invention, the chemical elements are consistent with the CAS version of the Periodic Table of Elements, and "Handbook of Chemistry and Physics", 75th Edition, 1994. In addition, the general principles of organic chemistry can refer to the description in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry" by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007 , The entire contents of which are incorporated herein by reference.

Unless otherwise stated or there is an obvious conflict in context, the articles "a", "an" and "said" used herein are intended to include "at least one" or "one or more." Therefore, the articles used herein refer to articles of one or more than one (ie at least one) object. For example, "a component" refers to one or more components, that is, more than one component may be considered to be adopted or used in the embodiment of the described embodiment.

The term "comprising" is an open-ended expression, that is, includes the content specified in the present invention, but does not exclude other aspects.

"Stereoisomers" refer to compounds that have the same chemical structure but differ in the arrangement of the atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotamers), geometric isomers (cis/trans) isomers, atropisomers, etc. .

"Chiral" refers to a molecule that can not overlap with its mirror image; and "achiral" refers to a molecule that can overlap with its mirror image.

"Enantiomers" refer to two isomers of a compound that cannot overlap but are mirror images of each other.

"Diastereoisomers" refer to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties and reactivity. Diastereomeric mixtures can be separated by high-resolution analytical procedures such as electrophoresis and chromatography, such as HPLC.

The stereochemistry definitions and rules used in the present invention generally follow SPParker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry" of Organic Compounds", John Wiley&Sons, Inc., New York, 1994.

Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to one or more of its chiral centers. The prefixes d and l or (+) and (-) are symbols used to specify the rotation of plane-polarized light caused by a compound, where (-) or l indicates that the compound is levorotatory. Compounds prefixed with (+) or d are dextrorotatory. A specific stereoisomer is an enantiomer, and a mixture of such isomers is called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which can occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

Any asymmetric atom (for example, carbon, etc.) of the compound disclosed in the present invention may exist in a racemic or enantiomerically enriched form, such as (R)-, (S)- or (R,S)-configuration form exist. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess.

Depending on the choice of starting materials and methods, the compounds of the present invention can be used as one of the possible isomers or their mixtures, such as racemates and diastereomeric mixtures (depending on the number of asymmetric carbon atoms). ) Exists in the form of. Optically active (R)- or (S)-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent may have an E or Z configuration; if the compound contains a disubstituted cycloalkyl, the substituent of the cycloalkyl may have a cis or trans configuration.

Any resulting mixture of stereoisomers can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physical and chemical properties of the components, for example, by chromatography Method and/or fractional crystallization method.

The racemate of any final product or intermediate obtained can be resolved into optical enantiomers by a method familiar to those skilled in the art using known methods, for example, by performing diastereomeric salts of the obtained diastereomers. Separate. The racemic product can also be separated by chiral chromatography, such as high performance liquid chromatography (HPLC) using a chiral adsorbent. In particular, enantiomers can be prepared by asymmetric synthesis, for example, refer to Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis (2 ^nd Ed. Robert E. Gawley, Jeffrey Aubé, Elsevier, Oxford, UK, 2012); Eliel, ELStereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, SHTables of Resolving Agents and Optical Resolutions p.268 (ELEliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972); Chiral Separation Techniques: A Practical Approach (Subramanian, G. Ed., Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2007).

The term "tautomer" or "tautomeric form" refers to structural isomers with different energies that can be converted into each other through a low energy barrier. If tautomerism is possible (as in solution), the chemical equilibrium of tautomers can be reached. For example, proton tautomers (also called prototropic tautomers) include interconversions through proton migration, such as keto-enol isomerization and imine-ene Amine isomerization. Valence tautomers include interconversion through the recombination of some bond-forming electrons. Specific examples of keto-enol tautomerism are the interconversion of pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-ketone tautomerism. A specific example of phenol-ketone tautomerism is the interconversion of pyridine-4-ol and pyridine-4(1H)-one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

As described in the present invention, the compounds of the present invention can be optionally substituted by one or more substituents, such as the compounds of the general formula above, or the special examples, subclasses, and subclasses included in the examples. A class of compounds. It should be understood that the term "optionally substituted" and the term "substituted or unsubstituted" can be used interchangeably. Generally speaking, the term "substituted" means that one or more hydrogen atoms in a given structure are replaced by a specific substituent. Unless otherwise indicated, an optional substituent group can be substituted at each substitutable position of the group. When more than one position in the given structural formula can be substituted by one or more substituents selected from specific groups, then the substituents can be substituted at each position with the same or different substitutions.

In addition, it should be noted that, unless explicitly pointed out in other ways, the description methods used in the present invention are interchangeable with "each ... independently being" and "... each independently being" and "... independently being". Should be understood in a broad sense, it can mean that the specific options expressed between the same symbols in different groups do not affect each other, or it can mean the specific options expressed between the same symbols in the same group Do not affect each other.

In each part of this specification, the substituents of the compounds disclosed in the present invention are disclosed according to the group type or scope. In particular, the present invention includes each independent sub-combination of each member of these group types and ranges. For example, the term "C1-6 alkyl" specifically refers to independently disclosed methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl.

Each of a, b, c, d, e, and f is each independently an integer between 0-30. In one embodiment, each of a, b, c, d, e, and f is each independently an integer between 0-25. In another embodiment, each of a, b, c, d, e, and f is each independently an integer between 0-20. In another embodiment, each of a, b, c, d, e, f is each independently an integer between 0-15. In another embodiment, each of a, b, c, d, e, and f is each independently an integer between 0-10. In another embodiment, each of a, b, c, d, e, and f is each independently an integer between 0-5.

In each part of the present invention, linking substituents are described. When the structure clearly requires a linking group, the Markush variables listed for the group should be understood as the linking group. For example, if the structure requires a linking group and the Markush group definition of the variable lists "alkyl" or "aryl", it should be understood that the "alkyl" or "aryl" respectively represents the attached Alkylene group or arylene group.

The term "alkyl" or "alkyl group" used in the present invention means a saturated linear or branched monovalent hydrocarbon group containing 1 to 20 carbon atoms, wherein the alkyl group may optionally Ground is substituted by one or more substituents described in this invention. Unless otherwise specified, alkyl groups contain 1-20 carbon atoms. In one embodiment, the alkyl group contains 1-12 carbon atoms; in another embodiment, the alkyl group contains 1-6 carbon atoms; in another embodiment, the alkyl group contains 1 -4 carbon atoms; in yet another embodiment, the alkyl group contains 1-3 carbon atoms.

Examples of alkyl groups include, but are not limited to, methyl (Me, -CH ₃ ), ethyl (Et, -CH ₂ CH ₃ ), n-propyl (n-Pr, -CH ₂ CH ₂ CH ₃ ), isopropyl (i-Pr, -CH(CH ₃ ) ₂ ), n-butyl (n-Bu, -CH ₂ CH ₂ CH ₂ CH ₃ ), isobutyl (i-Bu, -CH ₂ CH (CH ₃ ) ₂ ), sec-butyl (s-Bu, -CH(CH ₃ )CH ₂ CH ₃ ), tert-butyl (t-Bu, -C(CH ₃ ) ₃ ), n-pentyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl -2-butyl (-C(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl (-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1- Butyl (-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), n-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-Methyl-2-pentyl (-C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl (-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl (-CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl (-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl (-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl(-C(CH ₃ ) ₂ CH (CH ₃ ) ₂ ), 3,3-dimethyl-2-butyl (-CH(CH ₃ )C(CH ₃ ) ₃ ), n-heptyl, n-octyl, and so on.

The term "alkylene" refers to a saturated divalent hydrocarbon group obtained by removing two hydrogen atoms from a saturated linear or branched hydrocarbon group. Unless otherwise specified, alkylene groups contain 1-12 carbon atoms. In one embodiment, the alkylene group contains 1-6 carbon atoms; in another embodiment, the alkylene group contains 1-4 carbon atoms; in another embodiment, the alkylene group The group contains 1-3 carbon atoms; in yet another embodiment, the alkylene group contains 1-2 carbon atoms. Examples of this include methylene (-CH2-), ethylene (-CH2CH2-), isopropylidene (-CH(CH3)CH2-), and the like.

The term "alkenyl" means a linear or branched monovalent hydrocarbon group containing 2-12 carbon atoms, in which there is at least one site of unsaturation, that is, a carbon-carbon sp2 double bond, wherein the alkenyl group It may be optionally substituted by one or more substituents described in the present invention, including the positioning of "cis" and "tans", or the positioning of "E" and "Z". In one embodiment, the alkenyl group contains 2-8 carbon atoms; in another embodiment, the alkenyl group contains 2-6 carbon atoms; in yet another embodiment, the alkenyl group contains 2 -4 carbon atoms. Examples of alkenyl groups include, but are not limited to, vinyl (-CH=CH ₂ ), allyl (-CH ₂ CH=CH ₂ ), and the like.

The term "alkynyl" means a linear or branched monovalent hydrocarbon group containing 2-12 carbon atoms, in which there is at least one unsaturation site, that is, a carbon-carbon sp triple bond, wherein the alkynyl group It may be optionally substituted by one or more substituents described in the present invention. In one embodiment, the alkynyl group contains 2-8 carbon atoms; in another embodiment, the alkynyl group contains 2-6 carbon atoms; in yet another embodiment, the alkynyl group contains 2 -4 carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl (-C≡CH), propargyl (-CH ₂ C≡CH), 1-propynyl (-C≡C-CH ₃ ), etc. .

The term "heteroalkyl" means that one or more heteroatoms are inserted into the alkyl chain, wherein the alkyl group and the heteroatom have the meanings as described in the present invention. Unless otherwise specified, the heteroalkyl group contains 2-10 carbon atoms. In other embodiments, the heteroalkyl group contains 2-8 carbon atoms. In other embodiments, the heteroalkyl group contains 2 -6 carbon atoms, in other embodiments, the heteroalkyl group contains 2-4 carbon atoms, and in other embodiments, the heteroalkyl group contains 2-3 carbon atoms. Such examples include, but are not limited to, CH ₃ OCH ₂ -, CH ₃ CH ₂ OCH ₂ -, CH ₃ SCH ₂ -, (CH ₃ ) ₂ NCH ₂ -, (CH ₃ ) ₂ CH ₂ OCH ₂ -, CH ₃ OCH ₂ CH ₂ -, CH ₃ CH ₂ OCH ₂ CH ₂ -and so on.

The term "alkenylene" refers to an alkene group obtained by removing two hydrogen atoms from a linear or branched alkene. And the alkenylene group may be substituted or unsubstituted, wherein the substituent may be, but not limited to, deuterium, hydroxyl, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkyl, Alkenyl, alkynyl, heterocyclyl, mercapto, nitro or aryloxy. Such examples include, but are not limited to, vinylidene (-CH=CH-), isopropenylene (-C(CH3)=CH-), 3-methoxypropene-1,1-diyl, 2-methylbutene-1,1-diyl and so on.

The term "carbocyclylene" ("cycloalkylene") means a monocyclic ring containing 3-12 carbon atoms or a bicyclic ring containing 7-12 carbon atoms, which is a saturated divalent hydrocarbon ring obtained by removing two hydrogen atoms, Where carbocyclyl or cycloalkyl has the meaning as described in the present invention, such examples include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, 1-cyclopenta-1-ylidene Alkenyl, 1-cyclopent-2-alkenylene, etc.

The term "heterocyclylene" means a monocyclic, bicyclic or tricyclic ring system in which one or more atoms in the ring are independently selected from heteroatoms, and may be fully saturated or contain one or more unsaturations, but not It belongs to the aromatic group and has two connection points connected to the rest of the molecule, and the heterocyclic group has the meaning as described in the present invention. Such examples include, but are not limited to, piperidine-1,4-diyl, piperazine-1,4-diyl, tetrahydrofuran-2,4-diyl, tetrahydrofuran-3,4-diyl, aza Cyclobutane-1,3-diyl, pyrrolidine-1,3-diyl, etc.

The term "alkoxy" means that the alkyl group is connected to the rest of the molecule through an oxygen atom, where the alkyl group has the meaning as described in the present invention. Unless otherwise specified, the alkoxy group contains 1-12 carbon atoms. In one embodiment, the alkoxy group contains 1-6 carbon atoms; in another embodiment, the alkoxy group contains 1-4 carbon atoms; in another embodiment, the alkoxy group The group contains 1-3 carbon atoms. The alkoxy group may be optionally substituted with one or more substituents described in this invention.

Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH ₃ ), ethoxy (EtO, -OCH ₂ CH ₃ ), 1-propoxy (n-PrO, n- Propoxy, -OCH ₂ CH ₂ CH ₃ ), 2-propoxy (i-PrO, i-propoxy, -OCH(CH ₃ ) ₂ ), 1-butoxy (n-BuO, n- Butoxy, -OCH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl-l-propoxy (i-BuO, i-butoxy, -OCH ₂ CH(CH ₃ ) ₂ ), 2-but Oxygen (s-BuO, s-butoxy, -OCH(CH ₃ )CH ₂ CH ₃ ), 2-methyl-2-propoxy (t-BuO, t-butoxy, -OC(CH ₃ ) ₃ ), 1-pentyloxy (n-pentyloxy, -OCH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyloxy (-OCH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentoxy (-OCH(CH ₂ CH ₃ ) ₂ ), 2-methyl-2-butoxy (-OC(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butoxy Group (-OCH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butoxy (-OCH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butoxy (-OCH ₂ CH(CH ₃ )CH ₂ CH ₃ ), etc.

The term "haloalkyl", "haloalkenyl" or "haloalkoxy" means an alkyl, alkenyl or alkoxy group substituted with one or more halogen atoms. Examples of such include, but are not limited to, Trifluoromethyl, trifluoromethoxy, etc.

The term "hydroxyalkyl" and "hydroxy-substituted alkyl" means that an alkyl group is substituted with one or more hydroxy groups, wherein the alkyl group has the meaning described in the present invention. Such examples include, but are not limited to, hydroxymethyl, hydroxyethyl, 1,2-dihydroxyethyl, and the like.

The term "carbocyclic group" or "carbocyclic ring" means a monovalent or multivalent non-aromatic saturated or partially unsaturated monocyclic, bicyclic or tricyclic ring system containing 3-12 carbon atoms. Carbobicyclic groups include spirocarbonbicyclic groups and fused carbobicyclic groups, and suitable carbocyclic groups include, but are not limited to, cycloalkyl, cycloalkenyl, and cycloalkynyl. Examples of carbocyclic groups further include, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl- 3-alkenyl, cyclohexyl, 1-cyclohexyl-1-alkenyl, 1-cyclohexyl-2-alkenyl, 1-cyclohexyl-3-alkenyl, cyclohexadienyl, cycloheptyl, cyclooctyl Group, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, etc.

The term "cycloalkyl" means a monovalent or multivalent saturated monocyclic, bicyclic or tricyclic ring system containing 3-12 carbon atoms. In one embodiment, the cycloalkyl group contains 3-12 carbon atoms; in another embodiment, the cycloalkyl group contains 3-8 carbon atoms; in another embodiment, the cycloalkyl group contains 3-6 carbon atoms. carbon atom. The cycloalkyl group may be independently unsubstituted or substituted with one or more substituents described in the present invention.

基，二氮杂

基，硫氮杂

基，吲哚啉基，1,2,3,4-四氢异喹啉基、1,3-苯并二噁茂基、2-氧杂-5-氮杂双环[2.2.1]庚-5-基。杂环基中-CH2-基团被-C(O)-取代的实例包括，但不限于，2-氧代吡咯烷基、氧代-1,3-噻唑烷基、2-哌啶酮基、3,5-二氧代哌啶基和嘧啶二酮基。杂环基中硫原子被氧化的实例包括，但不限于，环丁砜基、1,1-二氧代硫代吗啉基。所述的杂环基基团可以任选地被一个或多个本发明所描述的取代基所取代。 The terms "heterocyclic group" and "heterocyclic ring" are used interchangeably herein, and both refer to a saturated or partially unsaturated monocyclic, bicyclic or tricyclic ring containing 3-12 ring atoms, wherein at least one ring atom is selected from Nitrogen, sulfur and oxygen atoms. Unless otherwise specified, the heterocyclic group may be a carbon group or a nitrogen group, and the -CH2- group may be optionally replaced by -C(O)-. The sulfur atom of the ring can optionally be oxidized to S-oxide. The nitrogen atom of the ring can optionally be oxidized to an N-oxygen compound. Examples of heterocyclic groups include, but are not limited to: oxiranyl, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl , Pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1,3-dioxanyl, disulfide ring Pentyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl , Dioxanyl, Dithianyl, Thioxanyl, Homopiperazinyl, Homopiperidinyl, Oxepanyl, Thiepanyl, Oxazepine

Base, diaza

Base, thiazepine

Group, indolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxanyl, 2-oxa-5-azabicyclo[2.2.1]heptan-5 -base. Examples of heterocyclic groups where the -CH2- group is substituted by -C(O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidinonyl , 3,5-dioxopiperidinyl and pyrimidinedione. Examples in which the sulfur atom in the heterocyclic group is oxidized include, but are not limited to, sulfolane and 1,1-dioxothiomorpholinyl. The heterocyclic group may be optionally substituted by one or more substituents described in the present invention.

基，二氮杂

基，硫氮杂

基。杂环基中-CH2-基团被-C(O)-取代的实例包括，但不限于，2-氧代吡咯烷基、氧代-1,3-噻唑烷基、2-哌啶酮基、3,5-二氧代哌啶基和嘧啶二酮基。杂环基中硫原子被氧化的实例包括，但不限于，环丁砜基、1,1-二氧代硫代吗啉基。所述的4-7个原子组成的杂环基基团可以任选地被一个或多个本发明所描述的取代基所取代。 In one embodiment, the heterocyclic group is a heterocyclic group consisting of 4-7 atoms, which refers to a saturated or partially unsaturated monocyclic ring containing 4-7 ring atoms, wherein at least one ring atom is selected from nitrogen and sulfur. And oxygen atoms. Unless otherwise specified, the heterocyclic group composed of 4-7 atoms may be a carbon group or a nitrogen group, and the -CH2- group may be optionally replaced by -C(O)-. The sulfur atom of the ring can optionally be oxidized to S-oxide. The nitrogen atom of the ring can optionally be oxidized to an N-oxygen compound. Examples of heterocyclic groups composed of 4-7 atoms include, but are not limited to: azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrroline Group, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1,3-dioxocyclopentyl, disulfide Cyclopentyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazine Group, dioxanyl, dithianyl, thiazinyl, homopiperazinyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepine

Base, diaza

Base, thiazepine

base. Examples of heterocyclic groups where the -CH2- group is substituted by -C(O)- include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidinonyl , 3,5-dioxopiperidinyl and pyrimidinedione. Examples in which the sulfur atom in the heterocyclic group is oxidized include, but are not limited to, sulfolane and 1,1-dioxothiomorpholinyl. The heterocyclic group composed of 4-7 atoms can be optionally substituted by one or more substituents described in the present invention.

In another embodiment, the heterocyclic group is a 4-atom heterocyclic group, which refers to a saturated or partially unsaturated monocyclic ring containing 4 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms Replaced. Unless otherwise specified, the 4-atom heterocyclic group may be a carbon group or a nitrogen group, and the -CH2- group may be optionally replaced by -C(O)-. The sulfur atom of the ring can optionally be oxidized to S-oxide. The nitrogen atom of the ring can optionally be oxidized to an N-oxygen compound. Examples of 4-atom heterocyclic groups include, but are not limited to: azetidinyl, oxetanyl, and thietanyl. The 4-atom heterocyclic group may be optionally substituted by one or more substituents described in the present invention.

In another embodiment, the heterocyclic group is a 5-atom heterocyclic group, which refers to a saturated or partially unsaturated monocyclic ring containing 5 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms . Unless otherwise specified, the 5-atom heterocyclic group may be a carbon group or a nitrogen group, and the -CH2- group may be optionally replaced by -C(O)-. The sulfur atom of the ring can optionally be oxidized to S-oxide. The nitrogen atom of the ring can optionally be oxidized to an N-oxygen compound. Examples of 5-atom heterocyclic groups include, but are not limited to: pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolinyl, Tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1,3-dioxanyl, dithiocyclopentyl. Examples of heterocyclic groups where the -CH2- group is substituted by -C(O)- include, but are not limited to, 2-oxopyrrolidinyl and oxo-1,3-thiazolidinyl. Examples in which the sulfur atom in the heterocyclic group is oxidized include, but are not limited to, a sulfolane group. The 5-atom heterocyclic group may be optionally substituted by one or more substituents described in the present invention.

In another embodiment, the heterocyclic group is a 6-atom heterocyclic group, which refers to a saturated or partially unsaturated monocyclic ring containing 6 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms . Unless otherwise specified, the 6-atom heterocyclic group may be a carbon group or a nitrogen group, and the -CH2- group may be optionally replaced by -C(O)-. The sulfur atom of the ring can optionally be oxidized to S-oxide. The nitrogen atom of the ring can optionally be oxidized to an N-oxygen compound. Examples of 6-atom heterocyclic groups include, but are not limited to: tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, Morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thiazinyl. Examples of the -CH2- group substituted by -C(O)- in the heterocyclic group include, but are not limited to, 2-piperidonyl, 3,5-dioxopiperidinyl, and pyrimidinedionyl. Examples in which the sulfur atom in the heterocyclic group is oxidized include, but are not limited to, 1,1-dioxothiomorpholinyl. The 6-atom heterocyclic group may be optionally substituted by one or more substituents described in the present invention.

In another embodiment, the heterocyclic group is a heterocyclic group composed of 7-12 atoms, which refers to a saturated or partially unsaturated spiro bicyclic or fused bicyclic ring containing 7-12 ring atoms, in which at least one ring atom Selected from nitrogen, sulfur and oxygen atoms. Unless otherwise specified, the heterocyclic group composed of 7-12 atoms may be a carbon group or a nitrogen group, and the -CH2- group may be optionally replaced by -C(O)-. The sulfur atom of the ring can optionally be oxidized to S-oxide. The nitrogen atom of the ring can optionally be oxidized to an N-oxygen compound. Examples of heterocyclic groups composed of 7-12 atoms include, but are not limited to: indolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,3-benzodioxenyl, 2- Oxa-5-azabicyclo[2.2.1]heptan-5-yl. The heterocyclic group composed of 7-12 atoms can be optionally substituted by one or more substituents described in the present invention.

The terms "fused bicyclic ring", "fused ring", "fused bicyclic group" and "fused ring group" are used interchangeably herein, and all refer to monovalent or multivalent saturated or partially unsaturated bridged ring systems, so The bridged ring system refers to a non-aromatic bicyclic ring system. Such a system may contain independent or conjugated unsaturated systems, but its core structure does not contain aromatic rings or aromatic heterocycles (but aromatic groups can be used as substituents on it).

The terms "spirocyclyl", "spirocyclic", "spirobicyclyl" or "spirobicyclic" are used interchangeably herein and refer to a monovalent or multivalent saturated or partially unsaturated ring system in which one ring originates from another A specific ring carbon atom on a ring. For example, as described below, a saturated bridged ring system (rings B and B') is called a "fused bicyclic ring", and ring A and ring B share a carbon atom in the two saturated ring systems and are It is called "spiro ring" or "spiro double ring". Each ring in the fused bicyclic group and the spiro bicyclic group may be a carbocyclic group or a heterocyclic group, and each ring is optionally substituted with one or more substituents described in the present invention.

The term "heterocycloalkyl" refers to a monovalent or multivalent saturated monocyclic, bicyclic or tricyclic ring system containing 3-12 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur or oxygen atoms.

The term "consisting of n atoms", where n is an integer, typically describes the number of ring atoms in a molecule, and the number of ring atoms in the molecule is n. For example, piperidinyl is a 6-atom heterocycloalkyl group, and 1,2,3,4-tetrahydronaphthalene is a 10-atom cycloalkyl group.

The term "unsaturated" as used in the present invention means that the group contains one or more degrees of unsaturation.

The term "heteroatom" refers to O, S, N, P and Si, including any oxidation state of N, S and P; primary, secondary, tertiary amine and quaternary ammonium salt forms; or on the nitrogen atom in the heterocyclic ring The form in which hydrogen is substituted, for example, N (like the N in 3,4-dihydro-2H-pyrrolyl), NH (like the NH in the pyrrolidinyl group) or NR (like the N-substituted pyrrolidinyl group NR).

The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I).

The term "aryl" means a monocyclic, bicyclic and tricyclic carbocyclic ring system containing 6-14 ring atoms, or 6-12 ring atoms, or 6-10 ring atoms, wherein at least one ring system is aromatic Family, where each ring system contains a ring composed of 3-7 atoms, and there are one or more attachment points connected to the rest of the molecule. The term "aryl" can be used interchangeably with the term "aromatic ring". Examples of aryl groups may include phenyl, naphthyl, and anthracene. The aryl group may be independently optionally substituted with one or more substituents described in the present invention.

The term "heteroaryl" refers to monocyclic, bicyclic and tricyclic ring systems containing 5-12 ring atoms, or 5-10 ring atoms, or 5-6 ring atoms, in which at least one ring system is aromatic, And at least one ring system contains one or more heteroatoms, wherein each ring system contains a ring composed of 5-7 atoms, and has one or more attachment points connected to the rest of the molecule. The term "heteroaryl" can be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic compound". The heteroaryl group is optionally substituted with one or more substituents described in the present invention. In one embodiment, the 5-10 atom heteroaryl group contains 1, 2, 3, or 4 heteroatoms independently selected from O, S, and N.

Examples of heteroaryl groups include, but are not limited to, 2-furyl, 3-furyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl , 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2- Pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (such as 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (such as 5-tetrazolyl), triazolyl (such as 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (such as 2-pyrazolyl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3- Triazolyl, 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5-thiodiazolyl, pyrazinyl, 1,3,5- Triazinyl; also includes the following bicyclics, but not limited to these bicyclics: benzimidazolyl, benzofuranyl, benzothienyl, indolyl (such as 2-indolyl), purinyl, quinolinyl (Such as 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), isoquinolinyl (such as 1-isoquinolinyl, 3-isoquinolinyl or 4-isoquinolinyl), imidazole And [1,2-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]Triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazole And [1,5-a]pyridyl, etc.

The term "carboxy", whether used alone or in combination with other terms, such as "carboxyalkyl", means -CO ₂ H; the term "carbonyl", whether used alone or in combination with other terms, such as "aminocarbonyl" or ""Acyloxy" means -(C=O)-.

The term "alkylamino" includes "N-alkylamino" and "N,N-dialkylamino" in which the amino groups are each independently substituted with one or two alkyl groups. In some examples, the alkylamino group is a _{lower alkylamino group with one or two Ci-6} alkyl groups attached to the nitrogen atom. In other embodiments, the alkylamino group is a C _1-3 lower alkylamino group. Suitable alkylamino groups can be monoalkylamino or dialkylamino. Examples of such include, but are not limited to, N-methylamino, N-ethylamino, N,N-dimethylamino, N,N -Diethylamino and so on.

The term "arylamino" means that the amino group is substituted with one or two aryl groups. Examples of this include, but are not limited to, N-phenylamino. In some examples, the aromatic ring on the arylamino group may be further substituted.

The term "aminoalkyl" includes C _1-10 straight or branched chain alkyl groups substituted with one or more amino groups. Some examples are that the aminoalkyl group is a C _1-6 "lower aminoalkyl group" substituted with one or more amino groups. Examples of this include, but are not limited to, aminomethyl, ammonia Ethyl, aminopropyl, aminobutyl and aminohexyl.

The term "prodrug" used in the present invention represents the conversion of a compound into a compound represented by formula (I) in vivo. Such conversion is affected by the hydrolysis of the prodrug in the blood or the enzymatic conversion of the prodrug into the maternal structure in the blood or tissues. The prodrug compounds of the present invention can be esters. In the existing invention, esters can be used as prodrugs including phenyl esters, aliphatic (C _1-24 ) esters, acyloxymethyl esters, and carbonates. , Carbamates and amino acid esters. For example, a compound in the present invention contains a hydroxyl group, that is, it can be acylated to obtain a compound in the form of a prodrug. Other prodrug forms include phosphate esters. For example, these phosphate ester compounds are obtained by phosphorylation of the parent hydroxyl group. For a complete discussion of prodrugs, refer to the following documents: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, J. Rautio et al., Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270, and SJ Hecker et al., Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51,2328-2345.

"Metabolite" refers to the product obtained by the metabolism of a specific compound or its salt in the body. The metabolites of a compound can be identified by techniques well known in the art, and its activity can be characterized by experimental methods as described in the present invention. Such products can be obtained by oxidizing, reducing, hydrolyzing, amidating, deamidating, esterifying, degreasing, enzymatic cleavage and the like of the administered compound. Correspondingly, the present invention includes the metabolites of the compound, including the metabolites produced by fully contacting the compound of the present invention with a mammal for a period of time.

The "pharmaceutically acceptable salt" used in the present invention refers to the organic and inorganic salts of the compound of the present invention. Pharmaceutically acceptable salts are well-known in the field, as described in the literature: SMBerge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66:1-19. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, inorganic acid salts formed by reaction with amino groups include hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, And organic acid salts such as acetate, oxalate, maleate, tartrate, citrate, succinate, malonate, or other methods described in books and literature such as ion exchange These salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphoric acid Salt, camphor sulfonate, cyclopentyl propionate, digluconate, lauryl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate Salt, gluconate, hemisulfate, heptanoate, caproate, hydroiodide, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, Malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectate, persulfate, 3 -Phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts obtained with appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. The present invention also contemplates the quaternary ammonium salt formed by any compound containing the N group. Water-soluble or oil-soluble or dispersed products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further comprise suitable amine cation nontoxic ammonium, quaternary ammonium, and the counterion, such as halide, hydroxide, carboxylate, sulfated, phosphorylated compounds, nitrate compounds, C _{1 -8} Sulfonates and aromatic sulfonates.

The "solvate" of the present invention refers to an association formed by one or more solvent molecules and the compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to the association formed by the solvent molecule being water.

The term "treating" any disease or condition as used in the present invention, in some embodiments refers to ameliorating the disease or condition (ie slowing down or preventing or reducing the development of the disease or at least one clinical symptom thereof). In other embodiments, "treating" refers to alleviating or improving at least one physical parameter, including physical parameters that may not be perceived by the patient. In other embodiments, "treatment" refers to the regulation of a disease or condition physically (e.g., stabilizing perceptible symptoms) or physiologically (e.g., stabilizing physical parameters) or both. In other embodiments, "treating" refers to preventing or delaying the onset, occurrence, or worsening of a disease or condition.

Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids, such as acetate, aspartate, benzoate, benzenesulfonate, bromide/hydrobromide, bicarbonate/ Carbonate, bisulfate/sulfate, camphorsulfonate, chloride/hydrochloride, chlorophylline, citrate, ethanedisulfonate, fumarate, glucoheptonate, glucose Saccharate, glucuronate, hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, maleate Acid salt, malonate, mandelate, methanesulfonate, methylsulfate, naphthoate, naphthalenesulfonate, nicotinate, nitrate, octadecanoate, oleate, oxalic acid Salt, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, polygalactonate, propionate, stearate, succinate, sulfosalicylate, tartrate , Tosylate and trifluoroacetate.

Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like.

Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid , Ethanesulfonic acid, p-toluenesulfonic acid, sulfosalicylic acid, etc.

Pharmaceutically acceptable base addition salts can be formed with inorganic bases and organic bases.

Inorganic bases from which salts can be derived include, for example, ammonium salts and metals from groups I to XII of the periodic table. In certain embodiments, the salt is derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium, and magnesium salts.

Organic bases from which salts can be derived include primary amines, secondary amines, and tertiary amines. Substituted amines include naturally occurring substituted amines, cyclic amines, and basic ion exchange resins. Certain organic amines include, for example, isopropylamine, benzathine, choline, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine .

The pharmaceutically acceptable salt of the present invention can be synthesized from the parent compound, basic or acidic moiety by conventional chemical methods. Generally speaking, such salts can be obtained by reacting the free acid form of these compounds with a stoichiometric amount of a suitable base (such as hydroxide, carbonate, bicarbonate, etc.) of Na, Ca, Mg or K, or by reacting These compounds are prepared by reacting the free base form of these compounds with a stoichiometric amount of a suitable acid. This type of reaction is usually carried out in water or an organic solvent or a mixture of the two. Generally, where appropriate, it is necessary to use a non-aqueous medium such as diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. For example, "Remington's Pharmaceutical Sciences", 20th Edition, Mack Publishing Company, Easton, Pa., (1985); and "Handbook of Pharmaceutical Salts: Properties, Selection, and Use)", a list of other suitable salts can be found in Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

In addition, the compounds disclosed in the present invention, including their salts, can also be obtained in the form of their hydrates or in the form of containing their solvents (for example, ethanol, DMSO, etc.), and used for their crystallization. The compounds disclosed in the present invention can form solvates inherently or by design with pharmaceutically acceptable solvents (including water); therefore, the present invention is intended to include solvated and unsolvated forms.

Any structural formula given in the present invention is also intended to represent the non-isotopically enriched form and the isotopically enriched form of these compounds. The isotope-enriched compound has the structure described by the general formula given in the present invention, except that one or more atoms are replaced by atoms having the selected atomic weight or mass number. Exemplary isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O , ¹⁸ O, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ Cl and ¹²⁵ I.

On the other hand, the compounds of the present invention include isotopically enriched compounds as defined in the present invention, for example, those compounds in which radioactive isotopes such as ³ H, ¹⁴ C and ¹⁸ F are present, or non-radioactive isotopes such as ² H and ¹³ C. Such isotopically-enriched compounds can be used for metabolism studies (using ¹⁴ C), reaction kinetics studies (using, for example, ² H or ³ H), detection or imaging techniques, such as positron emission tomography (PET) or including drugs or Single-photon emission computed tomography (SPECT), which measures the distribution of substrate tissue, may be used in radiotherapy of patients. ¹⁸ F-enriched compounds are particularly ideal for PET or SPECT research. The isotope-enriched compound of Formula I or Formula II can be prepared by conventional techniques familiar to those skilled in the art or as described in the examples and preparation process of the present invention, using a suitable isotope-labeled reagent instead of the previously used unlabeled reagent .

In addition, the substitution of heavier isotopes, particularly deuterium (ie, ² H or D), can provide certain therapeutic advantages due to higher metabolic stability. For example, the increase in the half-life in the body or the decrease in the dosage requirement or the improvement of the therapeutic index. The isotope enrichment factor can be used to define the concentration of such heavier isotopes, especially deuterium. If the substituent of the compound of the present invention is designated as deuterium, the compound has at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), for each designated deuterium atom, At least 4500 (67.5% deuterium doping), at least 5000 (75% deuterium doping), at least 5500 (82.5% deuterium doping), at least 6000 (90% deuterium doping), at least 6333.3 (95% deuterium doping) Deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation) isotope enrichment factor. The pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent may be isotopically substituted, such as D ₂ O, acetone-d ₆ , DMSO-d ₆ .

In another aspect, the present invention relates to an intermediate for preparing the compound contained in Formula I or Formula II.

In another aspect, the present invention relates to methods for the preparation, separation and purification of compounds contained in formula I or formula II.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention, a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle, or a combination thereof. In some embodiments, the pharmaceutical composition may be in liquid, solid, semi-solid, gel or spray form.

"Combination" means a fixed combination in a single dosage unit form or a kit of parts for combined administration, wherein the compound disclosed in the present invention and the combination partner can be administered independently at the same time or can be administered separately within a certain time interval, In particular, make joint partners show cooperation, such as synergy. The terms "co-administration" or "co-administration" and the like are intended to encompass the administration of the selected combination partner to a single individual (eg patient) in need thereof, and are intended to include treatment regimens in which the substances do not have to be administered via the same route of administration or simultaneously . The term "pharmaceutical combination" as used herein means a product obtained by mixing or combining more than one active ingredient, and includes both fixed and non-fixed combinations of active ingredients. The term "fixed combination" means that the active ingredients such as the compound disclosed in the present invention and the combination partner are simultaneously administered to the patient in the form of a single entity or dosage. The term "non-fixed combination" means that the active ingredients such as the compound disclosed in the present invention and the combination partner are both administered to the patient simultaneously, jointly or sequentially without a specific time limit as separate entities, wherein the administration provides a therapeutically effective level of the two compounds in the patient .

Unless otherwise specified, the experimental methods used in the following examples are all conventional methods.

The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

The Pomalidomide terminal derivatives used in the following examples were prepared according to the method disclosed in the literature Chemistry & Biology 22, 755-763 (2015). Lenalidomide terminal derivatives were prepared according to the method disclosed in the document J. Med. Chem (DOI: 10.1021/acs.jmedchem. 6b01816). The RG-7112 terminal carboxylic acid derivative was prepared according to the document Bioorg.Med.Chem.Lett.18,5904 -5908 (2008). and ACS Med. Chem. Lett. 4, 466-469 (2013).

The terminal derivatives of Ibrutinib used in the following examples were prepared according to the following method: The terminal alkynes required by Click chemistry were connected to the Ibrutinib intermediate (cas: 1022150-12-4) through an amide condensation reaction. The specific preparation process is as follows:

(1) Preparation of Intermediate 1a

In a 25 mL round bottom flask, 193 mg of Ibrutinib intermediate (cas: 1022150-12-4), 3 mL of anhydrous chloroform and 31 μL of propiolic acid were added. Dissolve 103 mg of DCC in 2 mL of anhydrous chloroform, keep the reaction solution at 0 degrees Celsius under stirring conditions, slowly add the anhydrous chloroform solution of DCC to the above reaction solution, and keep at 0 degrees Celsius and stir for 1 hour. Cool the reaction solution to -20°C, keep the filtrate after filtration, repeat the filtration twice, spin-dry the filtrate and use a 200-300 mesh silica gel chromatographic column for separation and purification. The mobile phase is dichloromethane: methanol = 60:1, that is The intermediate 1a was obtained with a yield of 72%.

Preparation of Intermediate 1b

In a 25 mL round-bottom flask, 193 mg of Ibrutinib intermediate (cas: 1022150-12-4), 3 mL of anhydrous chloroform and 42 mg of propioic acid were added. Dissolve 103 mg of DCC in 2 mL of anhydrous chloroform, keep the reaction solution at 0 degrees Celsius under stirring conditions, slowly add the anhydrous chloroform solution of DCC to the above reaction solution, and keep at 0 degrees Celsius and stir for 1 hour. Cool the reaction solution to -20°C, keep the filtrate after filtration, repeat the filtration twice, spin-dry the filtrate and use a 200-300 mesh silica gel chromatographic column for separation and purification. The mobile phase is dichloromethane: methanol = 60:1, that is The intermediate 1b was obtained with a yield of 62%.

Preparation of Intermediate 1c

Add 193mg Ibrutinib intermediate (cas: 1022150-12-4), 3mL anhydrous DMF, 75mg HOBT, 106mg EDCI, 49mg pentynoic acid, 77μL triethylamine and 10mg DMAP into a 25mL round bottom flask. The reaction solution was kept stirring at room temperature for 24 hours. The reaction was quenched by adding 15 mL saturated aqueous sodium chloride solution, the mixture was extracted three times with 15 mL×3 ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, spin-dried solvent, and separated using 200-300 mesh silica gel column For purification, the mobile phase is dichloromethane:methanol=40:1 to obtain intermediate 1c with a yield of 61%.

Preparation of Intermediate 1d

Add 193mg Ibrutinib intermediate (cas: 1022150-12-4), 3mL anhydrous DMF, 75mg HOBT, 106mg EDCI, 56mg hexynoic acid, 77μL triethylamine and 10mg DMAP into a 25mL round bottom flask. The reaction solution was kept stirring at room temperature for 24 hours. The reaction was quenched by adding 15 mL saturated aqueous sodium chloride solution, the mixture was extracted three times with 15 mL×3 ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, spin-dried solvent, and separated using 200-300 mesh silica gel column For purification, the mobile phase is dichloromethane:methanol=40:1 to obtain intermediate 1d with a yield of 71%.

(2) Preparation of Intermediate 2a

In a 100 mL round bottom flask, 2.67 mL 3-aminopropan-1-ol and 50 mL anhydrous dichloromethane were added, the reaction solution was cooled to 0 degrees Celsius, and 6.8 mL triethylamine and 9.7 mL Boc ₂ O were added. The reaction solution was naturally returned to room temperature under stirring conditions, and stirring was continued for 12 hours. The reaction was quenched by adding 100 mL saturated sodium bicarbonate aqueous solution, the mixture was extracted three times with 50 mL×3 dichloromethane, the organic phases were combined, dried with anhydrous sodium sulfate, spin-dried solvent, and separated using 200-300 mesh silica gel column For purification, the mobile phase is dichloromethane:methanol=20:1 to obtain Intermediate 2a with a yield of 72%.

Preparation of Intermediate 2b

In a 100 mL round bottom flask, 3.22 mL 4-aminobutan-1-ol and 50 mL anhydrous dichloromethane were added, the reaction solution was cooled to 0 degrees Celsius, and 6.8 mL triethylamine and 9.7 mL Boc ₂ O were added. The reaction solution was naturally returned to room temperature under stirring conditions, and stirring was continued for 12 hours. The reaction was quenched by adding 100 mL saturated sodium bicarbonate aqueous solution, the mixture was extracted three times with 50 mL×3 dichloromethane, the organic phases were combined, dried with anhydrous sodium sulfate, spin-dried solvent, and separated using 200-300 mesh silica gel column For purification, the mobile phase is dichloromethane:methanol=20:1 to obtain Intermediate 2b with a yield of 81%.

Preparation of Intermediate 2c

Add 1.6 g of sodium azide and 25 mL of water into a 100 mL round bottom flask, and add 4.9 g of 2-(2-hydroxyethoxy)ethyl-4-methylbenzenesulfonate. The reaction solution was stirred at 90 degrees Celsius for 24 hours. The reaction was quenched by adding 50 mL saturated aqueous sodium bicarbonate solution, the mixture was extracted three times with 40 mL×3 dichloromethane, the organic phases were combined, dried with anhydrous sodium sulfate, and the solvent was spin-dried to obtain the intermediate with a yield of 46%. Add 787 mg of the product obtained, 8 mL of anhydrous dichloromethane and 1.67 mL of triethylamine into a 50 mL round bottom flask, and slowly add 1.72 g of TsCl anhydrous dichloromethane solution to the above reaction solution under stirring conditions. Stir for 24 hours. The reaction was quenched by adding 100 mL saturated sodium bicarbonate aqueous solution, the mixture was extracted three times with 50 mL×3 dichloromethane, the organic phases were combined, dried with anhydrous sodium sulfate, spin-dried solvent, and separated using 200-300 mesh silica gel column For purification, the mobile phase is petroleum ether: ethyl acetate = 5:1, and the intermediate 2c is obtained with a yield of 67%.

Preparation of intermediate 2d

Add 526mg of intermediate 2a and 15mL of anhydrous THF to a 50mL round bottom flask, slowly add 150mg of NaH (60% dispersed in mineral oil) under stirring at 0 degrees Celsius, and stir the reaction solution at 0 degrees Celsius for 1 hour. An anhydrous THF solution of 285 mg of intermediate 2c was slowly added to the above reaction solution, and the reaction solution was naturally returned to room temperature under stirring conditions, and stirring was continued for 15 hours. Methanol was added dropwise to the reaction solution to quench the reaction, the solvent was spin-dried, and the 200-300 mesh silica gel column was used for separation and purification. The mobile phase was petroleum ether: ethyl acetate = 6:1, and the intermediate was obtained. The yield was 26%. . In a 25 mL round bottom flask were added 230 mg of the obtained product, 5 mL of dichloromethane and 0.5 mL of trifluoroacetic acid, and stirred at room temperature for 6 hours. Add 15 mL of toluene, spin dry the solvent, and use a 200-300 mesh silica gel column for separation and purification. The mobile phase is dichloromethane: methanol = 10:1 to obtain intermediate 2d with a yield of 62%.

Preparation of Intermediate 2e

Add 2.8g intermediate 2b and 50mL anhydrous THF into a 100mL round bottom flask, slowly add 800mg NaH (60%, dispersed in mineral oil) under stirring at 0 degrees Celsius, and stir the reaction solution at 0 degrees Celsius for 1 hour. An anhydrous THF solution of 1.43 g of intermediate 2c was slowly added to the above reaction solution, and the reaction solution was naturally returned to room temperature under stirring conditions, and stirring was continued for 15 hours. Methanol was added dropwise to the reaction solution to quench the reaction, the solvent was spin-dried, and the 200-300 mesh silica gel column was used for separation and purification. The mobile phase was petroleum ether: ethyl acetate = 6:1, and the intermediate was obtained. The yield was 22%. . In a 25 mL round bottom flask were added 800 mg of the obtained product, 15 mL of dichloromethane and 1.5 mL of trifluoroacetic acid, and stirred at room temperature for 2 hours. Add 25 mL of toluene, spin dry the solvent, and use a 200-300 mesh silica gel chromatographic column for separation and purification. The mobile phase is dichloromethane: methanol = 10:1 to obtain Intermediate 2e with a yield of 66%.

(3) Preparation of Intermediate 4a

Add 24 mg 8-aminooctanoic acid, 70 μL DIEA, 0.3 mL DMF, and 35 mg 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione to a 5 mL round bottom flask. Keep the reaction solution stirred at 90 degrees Celsius for 3 hours. The reaction was quenched by adding 5 mL saturated sodium chloride aqueous solution, the mixture was extracted three times with 5 mL×3 ethyl acetate, the organic phases were combined, dried with anhydrous sodium sulfate, spin-dried solvent, and separated using 200-300 mesh silica gel column For purification, the mobile phase is dichloromethane:methanol=40:1 to obtain Intermediate 4a with a yield of 22%.

Preparation of Intermediate 4b

Add 26mg 9-aminooctanoic acid, 70μL DIEA, 0.3mL DMF, and 35mg 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione into a 5mL round bottom flask. Keep the reaction solution stirred at 90 degrees Celsius for 3 hours. The reaction was quenched by adding 5 mL saturated sodium chloride aqueous solution, the mixture was extracted three times with 5 mL×3 ethyl acetate, the organic phases were combined, dried with anhydrous sodium sulfate, spin-dried solvent, and separated using 200-300 mesh silica gel column For purification, the mobile phase is dichloromethane:methanol=40:1 to obtain Intermediate 4b with a yield of 31%.

(4) Preparation of Intermediate 5a

Add 3g octane-1,8-diol, 20mL anhydrous THF and 1.3g potassium tert-butoxide into a 100mL round bottom flask. The reaction solution was kept stirring for 15 minutes under argon protection at room temperature, and 1.2 g of 3-bromopropyne was added dropwise. The reaction solution was kept stirred for 12 hours under argon protection at room temperature. Rotate the solvent to dryness, add 30mL saturated sodium chloride aqueous solution to quench the reaction, adjust the pH to 3-5 with 1M HCl, extract the mixture three times with 30mL×3 dichloromethane, combine the organic phases, and use anhydrous sodium sulfate Dry, spin off the solvent, and use a 200-300 mesh silica gel chromatographic column for separation and purification. The mobile phase is petroleum ether: ethyl acetate = 3:1 to obtain Intermediate 5a with a yield of 55%.

Preparation of intermediate 5b

In a 100 mL round bottom flask, 12.6 g of 2,2'-(ethane-1,2-diylbis(oxy))bis(ethan-1-ol), 50 mL of anhydrous THF and 5 g of potassium tert-butoxide were added. The reaction solution was kept stirring for 15 minutes under argon protection at room temperature, and 3.5 mL of 3-bromopropyne was added dropwise. The reaction solution was kept stirred for 12 hours under argon protection at room temperature. Rotate the solvent to dryness, add 30mL saturated sodium chloride aqueous solution to quench the reaction, adjust the pH to 3-5 with 1M HCl, extract the mixture three times with 30mL×3 dichloromethane, combine the organic phases, and use anhydrous sodium sulfate Dry, spin off the solvent, and use a 200-300 mesh silica gel chromatographic column for separation and purification. The mobile phase is petroleum ether: ethyl acetate = 3:1 to obtain Intermediate 5b with a yield of 77%.

Preparation of intermediate 5c

Add 4.4mL pentane-1,5-diol, 30mL anhydrous THF and 2.5g potassium tert-butoxide into a 100mL round bottom flask. The reaction solution was kept under argon protection and room temperature and stirred for 15 minutes, and 1.73 mL of 3-bromopropyne was added dropwise. The reaction solution was kept stirred for 12 hours under argon protection at room temperature. Rotate the solvent to dryness, add 30mL saturated sodium chloride aqueous solution to quench the reaction, adjust the pH to 3-5 with 1M HCl, extract the mixture three times with 30mL×3 dichloromethane, combine the organic phases, and use anhydrous sodium sulfate Dry, spin off the solvent, and use a 200-300 mesh silica gel chromatographic column for separation and purification. The mobile phase is petroleum ether: ethyl acetate = 3:1 to obtain Intermediate 5c with a yield of 74%.

(4) Preparation of Intermediate 6a

In a 25mL round-bottom flask, add 276mg of lithium acetylene ethylenediamine complex dissolved in 8mL DMSO, put it in an ice bath to cool to 0℃, dissolve 6-bromohexanoic acid in 8mL DMSO, and slowly add dropwise to the reaction solution. Continue to stir in the bath for 10 minutes, slowly rise to room temperature and stir for 2 hours. The reaction solution was poured into ice water, acidified with 1M hydrochloric acid, and the mixture was extracted three times with 30 mL×3 dichloromethane. The organic phases were combined, dried with anhydrous sodium sulfate, and the solvent was spin-dried to dryness using a 200-300 mesh silica gel column. After separation and purification, the mobile phase is petroleum ether: ethyl acetate = 10:1, and the intermediate 6a is obtained with a yield of 35%.

Preparation of Intermediate 6b

Add 7.3g methyl 3-bromo-2-methylbenzoate, 6.5g NBS and 529mg AIBN to a 250mL round bottom flask, add 100mL dry chloroform to dissolve and reflux at 90°C for 5 hours. After the solvent was spin-dried, 60 mL of saturated sodium chloride aqueous solution was added to quench the reaction, the mixture was extracted three times with 60 mL×3 dichloromethane, the organic phases were combined, dried with anhydrous sodium sulfate, and the solvent was spin-dried to obtain an intermediate. 7.6 g of the obtained intermediate was mixed with 5.3 g of 2,6-Dioxopiperidine-3-ammonium chloride, 4.6 mL of Et ₃ N, 90 mL of dry acetonitrile was added, and the reaction solution was stirred at 80° C. for 10 hours. The solvent is spin-dried, and a 200-300 mesh silica gel column is used for separation and purification. The mobile phase is dichloromethane: methanol = 20:1 to 5:1 to obtain Intermediate 6b with a yield of 60%.

Preparation of Intermediate 6c

In a 25 mL round bottom flask were added 35 mg of Intermediate 6b, 30 mg of Intermediate 6a, 5 mg of cuprous iodide, 15 mg of Pd(dppf) ₂ Cl ₂ , 6 mL of DMF and 3 mL of Et ₃ N, and reacted at 80° C. for 3 hours under nitrogen protection. The solvent was spin-dried, and the 200-300 mesh silica gel column was used for separation and purification. The mobile phase was dichloromethane: methanol = 25:1 to obtain Intermediate 6c with a yield of 82%.

Preparation of Intermediate 6d

In a 25 mL round bottom flask were added 35 mg of Intermediate 6b, 30 mg of Intermediate 6a, 5 mg of cuprous iodide, 15 mg of Pd(dppf) ₂ Cl ₂ , 6 mL of DMF and 3 mL of Et ₃ N, and reacted at 80° C. for 3 hours under nitrogen protection. The solvent was spin-dried, and a 200-300 mesh silica gel column was used for separation and purification, and the mobile phase was dichloromethane: methanol = 25:1 to obtain Intermediate 6d with a yield of 81%.

Example 1. Preparation of compounds represented by formula 1 and formula 18

In a 10 mL round bottom flask, 50 mg of Ibrutinib intermediate (cas: 1022150-12-4), 56 mg of intermediate 4b, 21 mg of HOBT, 30 mg of EDCI, 20 μl of Et ₃ N, 2 mg of DMAP, and 2 ml of DMF were added to the flask and stirred at room temperature for 12 hours. Add 10mL saturated sodium chloride aqueous solution to the reaction solution, extract the mixture three times with 10mL×3 ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, spin off the solvent, and use 200-300 mesh silica gel column for separation and purification , The mobile phase is dichloromethane:methanol=40:1, formula 10 is obtained, and the yield is 85%.

In a 10 mL round bottom flask, 50 mg of Ibrutinib intermediate (cas: 1022150-12-4), 54 mg of intermediate 4a, 21 mg of HOBT, 30 mg of EDCI, 20 μl of Et ₃ N, 2 mg of DMAP, and 2 ml of DMF were added to the flask and stirred at room temperature for 12 hours. Add 10mL saturated sodium chloride aqueous solution to the reaction solution, extract the mixture three times with 10mL×3 ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, spin off the solvent, and use 200-300 mesh silica gel column for separation and purification , The mobile phase is dichloromethane:methanol=40:1, formula 12 is obtained, and the yield is 86%.

In a 10 mL round bottom flask were added 30 mg of Ibrutinib intermediate (cas: 1022150-12-4), 30 mg of intermediate 6d, 13 mg of HOBT, 18 mg of EDCI, 15 μl of Et ₃ N, 6 mg of DMAP, and 2 ml of DMF and stirred at room temperature for 12 hours. Add 10mL saturated sodium chloride aqueous solution to the reaction solution, extract the mixture three times with 10mL×3 ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, spin off the solvent, and use 200-300 mesh silica gel column for separation and purification The mobile phase is dichloromethane:methanol=30:1 to obtain the intermediate. The intermediate is dissolved in 4mL methanol and 1mL DMF, 20mg palladium carbon is added, and the reaction is carried out in hydrogen at 38°C for 8 hours. The reaction solution was filtered with diatomaceous earth, the solvent was spin-dried, and the 200-300 mesh silica gel column was used for separation and purification. The mobile phase was dichloromethane:methanol=30:1, and the formula 16 was obtained, and the yield was 72%.

In a 10 mL round bottom flask were added 30 mg of Ibrutinib intermediate (cas: 1022150-12-4), 30 mg of intermediate 6d, 13 mg of HOBT, 18 mg of EDCI, 15 μl of Et ₃ N, 6 mg of DMAP, and 2 ml of DMF and stirred at room temperature for 12 hours. Add 10mL saturated sodium chloride aqueous solution to the reaction solution, extract the mixture three times with 10mL×3 ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, spin off the solvent, and use 200-300 mesh silica gel column for separation and purification The mobile phase is dichloromethane:methanol=30:1 to obtain the intermediate. The intermediate is dissolved in 4mL methanol and 1mL DMF, 20mg palladium carbon is added, and the reaction is carried out in hydrogen at 38°C for 8 hours. The reaction solution was filtered with diatomaceous earth, the solvent was spin-dried, and the 200-300 mesh silica gel column was used for separation and purification. The mobile phase was dichloromethane:methanol=30:1, and the formula 18 was obtained, and the yield was 69%.

Example 2. The biological activity test of the compound at the Western Blot level

Cell processing (wild-type cells):

Set up Ramos or HBL-1 or Mino or IgE MM or Jurkat or HeLa cells in logarithmic growth phase in a 6-well plate and add compound treatment; harvest the cells after 48h.

Cell processing (transient cells with C481S or C481A mutant BTK plasmid):

Set up logarithmic growth phase of 293T or HeLa cells in a 6-well plate. After 18 hours, replace the culture medium after the cells adhere to the wall. After 1.5 hours, add 500 μl of freshly prepared Opti-MEM medium containing PEI and plasmid to each well. (The preparation method of Opti-MEM medium containing PEI and plasmid is shown below), mix well, replace normal culture medium and add compound after 8 hours, treat for 48 hours; change medium and add compound, and harvest cells after another 48 hours .

Preparation method of Opti-MEM medium containing PEI and plasmid:

Add 0.2 μg or 0.5 μg plasmid to 250 μl pre-warmed Opti-MEM medium, shake gently to mix and let stand for 5 minutes; add 0.6 μg or 1.5 μg PEI to 250 μl pre-warmed Opti-MEM culture After mixing, gently shake and let it stand for 5 minutes; slowly add the Opti-MEM medium containing PEI to the Opti-MEM medium containing the plasmid, shake gently while adding, and let it stand for 30 minutes.

Cell protein extraction:

Collect the cells: scrape the treated cells in the culture medium, fully suspend the cells and collect them by centrifugation at 300g for 5 minutes. After washing with PBS, discard the PBS.

Cell lysis: Add 150μl of 2×Loading Buffer to each sample, shake and mix thoroughly, denature at 95°C for 15 minutes, mix well and store at -20°C or use it directly for Western Blot detection.

The formula of 5×Loading Buffer is: 250mM Tris-HCl(pH6.8), 10%(W/V)SDS, 0.5%(W/V) bromophenol blue, 50%(V/V) glycerin, 5%( W/V) β-Mercaptoethanol (2-ME). 2×Loading Buffer is prepared by adding 1.5 times the volume of dd water to 5×Loading Buffer.

The specific steps of Western Blot detection are as follows:

1) Prepare an appropriate concentration of SDS-PAGE gel. Refer to Table 18.3 on page 883 of "Molecular Cloning Experiment Guide" (Science Press, Second Edition) to prepare a separation gel of suitable concentration, and refer to Table 18.4 on page 883 to prepare a concentrated gel with a concentration of 4%.

2) Prepare samples. Prepare protein samples according to the experimental requirements, denature them at 95°C for 15 minutes, centrifuge, mix and load them into the sample wells of the SDS-PAGE gel. Adjust the sample volume appropriately according to the protein quantification results, usually 4μl per well.

3) Electrophoresis. When the power is turned on, the voltage of the protein sample in the concentrated gel is 83 volts. When the protein sample enters the separation gel, we adjust the voltage to 110 volts to continue electrophoresis. Stop the electrophoresis when the bromophenol blue almost completely runs out of the PAGE gel.

4) Transfer film. After the electrophoresis, remove the gel and install the transfer device in the following order: (negative electrode), filter paper, gel, activated PVDF membrane, filter paper, (positive electrode). Remember that there must be no air bubbles between the gel and the PVDF membrane. Then the clamping and transferring device is placed in the transfer buffer solution, and finally placed in the ice box, placed in a 4°C cold storage, and energized at 100V constant voltage for 1.5 hours.

5) Closed. After the transfer, the PVDF membrane was taken out, and the membrane was immersed in TBST buffer containing 5% skimmed milk powder, and shaken on a shaker for 1 hour at room temperature.

6) Primary antibody incubation. After blocking, shake and wash 3 times with TBST buffer, then add a moderately diluted primary antibody at 4°C overnight. Recover the primary antibody, and wash the PVDF membrane with TBST buffer 3 times, shaking for 10 minutes each time.

7) Incubation with secondary antibody. Discard the TBST buffer, add a certain dilution ratio (usually 1:3000 to 1:5000) of the secondary antibody (mouse antibody or rabbit antibody, depending on the primary antibody), shake at room temperature for 1 hour. The secondary antibody was discarded, and the PVDF membrane was shaken and washed 3 times with TBST buffer for 10 minutes each time. Finally, wash with TBST buffer for 10 minutes.

8) Color development and compression. The ECL color developing substrate is evenly covered on the PVDF film, and the color is developed at room temperature for 0.5-15 minutes.

The degradation activity of the compounds according to the embodiments of the present invention on BTK is as follows:

The compound according to the embodiment of the present invention has a strong effect on the degradation of wild-type BTK, and can detect obvious degradation under 10-50 nM, and has no inhibitory or degradation effect on other targets such as EGFR, ITK, TEC, etc., and has specific targeting The effect of degrading BTK protein, some of the results are shown in Figure 4, where L18I represents the compound

Example 3. Inhibition of compound on BTK, ITK, EGFR

1. Prepare 1x kinase matrix buffer and stop buffer.

1) 1x Kinase Matrix Buffer

50mM HEPES, pH 7.5

0.01% Brij-35

2) Stop buffer

100mM HEPES, pH 7.5

0.015% Brij-35

0.2％Coating Reagent#3

50mM EDTA

2. Prepare the compound.

1) Use 100% DMSO to dilute the compound to reach 50X of the highest concentration of inhibitor required in the final reaction. If the compound is tested at 1 μM, prepare a 50 μM DMSO solution of the compound in this step.

2) The compound was serially diluted in a 3-fold gradient to obtain a total of 10 concentrations.

3) Add 100 microliters of 100% DMSO to the 2 empty wells of the 96-well plate as a no compound control and no enzyme control.

4) Prepare the middle plate.

Transfer 10 microliters of compound from the source plate to a new 96-well plate to serve as an intermediate plate.

Add 90 μl of 1x Kinase Buffer to each well of the middle plate.

Mix the compound in the middle plate on a shaker for 10 minutes

3. Prepare the enzyme-linked plate.

1) Transfer 5 microliters from each well of the 96-well middle plate to a 384-well plate in duplicate. For example, the 96-well plate A1 is transferred to the 384-well plate A1 and A2, the 96-well plate A2 is transferred to the 384-well plate A3 and A4, and so on.

4. Kinase reaction.

1) Prepare 2.5x enzyme solution

Add enzyme to 1x Kinase Matrix Buffer.

2) Prepare 2.5x peptide solution

Add FAM-labeled peptide and ATP to 1x kinase matrix buffer.

3) Transfer the 2.5x enzyme solution to the enzyme-linked plate.

4) The ELISA plate already contains 5 microliters of compound in 10% DMSO.

5) Add 10 microliters of 2.5x enzyme solution to each well of the 384-well ELISA plate.

6) Incubate at room temperature for 10 minutes.

7) Transfer the 2.5x peptide solution to the enzyme-linked plate.

Add 10 microliters of 2.5x peptide solution to each well of the 384-well ELISA plate.

8) Kinase reaction and stop

Incubate at 28 degrees Celsius for the specified time.

Add 25 μl of stop buffer to stop the reaction.

9) Collect data on Caliper.

5. Curve fitting

1) Copy the conversion data from the Caliper program.

2) Convert the conversion value to a suppression value.

Inhibition rate=(max-conversion value)/(max-min)*100.

"Max" stands for DMSO control; "min" stands for low control.

3) Fit the data in XLfit excel add-in version 5.4.0.8 to get the IC ₅₀ value.

The equation used is

Y=Bottom+(Top-Bottom)/(1+(LogIC ₅₀ /X)*HillSlope))

Ibrutinib BTK inhibitory activity of an IC ₅₀ of 1.4nM; Ibrutinib ITK inhibitory activity of an IC ₅₀ of 34nM, EGFR inhibitory activity of IC ₅₀ of 1.3nM. The compounds according to the embodiments of the present invention have an IC ₅₀ of 10-100 nM for inhibitory activity against BTK kinase, and the IC ₅₀ values for ITK and EGFR are both higher than 1000 nM, indicating that the compounds according to the embodiments of the present invention have no obvious side effects on Ibrutinib. Inhibition.

Example 4. Inhibition of cell proliferation by compounds represented by formula 1 to formula 18

MTT test reagents:

Reagents: RPIM 1640 medium; DMEM medium; 100× non-essential amino acids (NEAA); 100× penicillin streptomycin mixture; 50 mM β-mercaptoethanol; calf serum (FBS, previously inactivated).

Medium A (500ml): RPIM 1640 medium (450ml)+100×NEAA (5ml)+100×penicillin mixture (5ml)+calf serum (50ml)+50mM β-mercaptoethanol (0.5ml).

Medium B (500ml): DMEM medium (450ml) + 100 x NEAA (5ml) + 100 x penicillin mixture (5ml) + calf serum (50ml) + 50mM β mercaptoethanol (0.5ml). .

CCK-8 Kit (Cell Counting Kit-8)

MTT experiment process (Ramos cell and HBL-1cell):

1) Collect log phase cells and adjust the cell suspension concentration to 6.6×10 ⁴ /ml with medium A.

2) Use medium A to dilute the small molecules in a 2-fold gradient from 100 nM to 2 nM. Configure into a small molecule solution.

3) Add 45 μL of cell suspension to a 96-well plate (the edge holes are filled with sterile PBS, 3000 cells/well). A negative control (45 μL of cell suspension and 45 μL of A medium) was set for each plate, and 3 replicate wells were set for each group.

4) After incubating at 37°C and 5% CO ₂ for 1 hour, add 45 μL of the corresponding small molecule solution to each well of the 96-well plate. Then incubate at 37°C and 5% CO ₂ for 72-96 hours. In the system, the concentration of small molecules was diluted by a 2-fold gradient, ranging from 50 nM to 1 nM.

Add 10μl cck-8 solution to each well and continue to incubate for 4h. Direct enzyme-linked immunoassay OD490nm measures the absorbance of each well.

The results of inhibition of cell proliferation by the compounds according to the examples of the present invention measured by the above methods are shown in Table 1.

Table 1: MTT experiment (GI50 value) of the inhibitory ability of compounds represented by formula 1 to formula 18 on HBL-1 cell line and Ramos cell line:

Among them, N.D means that no inhibitory activity was detected.

It can be seen from Table 1 above that the inhibitory ability of the compounds according to the examples of the present invention on the proliferation of HBL-1 cells is equivalent to or even better than that of Ibrutinib.

Example 5. Inhibition of the compound on the release of inflammatory factors by THP-1 upon LPS stimulation

1. Prepare LPS.

Add FBS-free RPMI1640 medium to the LPS solid powder, the final concentration is 1mg/ml, and store in aliquots.

2. Prepare the compound (see Table 2 for compound structure and number).

1) Use 100% DMSO to dilute the compound to reach 100X of the highest concentration of inhibitor required in the reaction. If the compound is tested at 100 nM, prepare a 10 μM DMSO solution of the compound in this step.

2) Dilute the compound in the culture medium at a ratio of 100 times.

3. Drug treatment of cells.

1) Dilute THP-1 cells to the required concentration, inoculate 1X10^6 cells in each well of the six-well plate, and supplement the medium to 2ml.

2) Take 500ul of diluted drug and DMSO control in the corresponding wells, and set a blank control group without drug and DMSO at the same time, and incubate for 24h in a 37°C incubator. Repeat 3 times for each group.

4. LPS stimulation.

Take the corresponding volume of 1mg/ml LPS solution, add the medium to the six-well plate after diluting it to a concentration of 0.5ug/ml, and incubate for 24 hours.

5. RT-PCR to detect the level of inflammatory factors

1) Collect the treated THP-1 cells, centrifuge at 100g for 5 minutes, wash twice with PBS, extract the total RNA of the cells using the Trizol method, and then synthesize cDNA with CWBIO EasyQuick RT MasterMix as a template for qPCR.

2) PCR primers designed to detect IL-1β, TNFα, IL-6, IL-10 and internal reference GAPDH are designed according to the gene sequence of human pro-inflammatory cytokines, and the designed primers are used for fluorescence quantitative PCR (see table for specific conditions) 3).

The results of inhibition of the release of cellular inflammatory factors by the compounds of the present invention measured by the above methods are shown in Figure 3 and Table 4. The exemplary compounds have different RNA levels of IL-1β, TNFα, IL-6, and IL-10 in cells. The degree of down-regulation, in which the compound represented by formula 10 inhibits the release of inflammatory factors is the most obvious, indicating that the compounds of the present invention can inhibit the BTK pathway and thereby inhibit the release of inflammatory factors.

Table 2: Compound structure and number

Table 3: RT-PCR

Among them, target gene refers to target gene, species refers to species, primer name refers to the name of the guide, and sequence refers to the sequence of the guide.

Table 4: Compounds' regulation of inflammatory factor mRNA levels

To P9I L8I mL8I IL-1β down down down TNFα down down down

IL-6 down down down IL-10 down down down

Among them, down means down.

Example 6. Effects of compounds in mouse arthritis and pulmonary hemorrhage models

1. Arthritis

Experimental program: Fully emulsify bovine type II collagen (CII) with an equal volume of complete Freund's adjuvant (CFA) to prepare an emulsion containing 1 mg/mL CII. Mice in each group were injected with 100μL at the root of the tail under aseptic conditions. After the model was successfully induced, the mice were randomly divided into 4 groups, namely, the control group, the ibrutinib group, the 50mg/kg exemplary compound and 100mg/kg exemplary compound Group, and injected the corresponding drugs respectively. The arthritis index (AI) scores for each group of mice every day: 0 points, normal; 1 point, erythema and slight swelling of the ankle joint; 2 points, erythema and slight swelling of the ankle joint to the plantar joint or palm ; 3 points, erythema and moderate swelling from the ankle joint to the metatarsophalangeal joint or palm joint; 4 points, erythema and severe swelling from the ankle joint to the toe joint; the maximum score per mouse is 16 points.

Experimental results: Ibrutinib can significantly improve the symptoms of foot swelling in mice. Compared with the blank group, the example compound can improve the symptoms of arthritis.

2. Pulmonary hemorrhage

Experimental protocol: B6 mice were divided into healthy group (3 mice), blank group (7 mice), ibrutinib group (7 mice) and example compound group (7 mice). Except for the healthy group, all mice were injected intraperitoneally with 1ml at a time Norphytane, the blank group, ibrutinib group and the example compound were injected with vehicle, ibrutinib and the example compound respectively one week before the administration of norphytane, intraperitoneal injection at 100 mg/kg per day, and continuous injection after the injection of norphytane The mice were sacrificed 18 days after the injection, the lungs and spleen were taken out, weighed, and the bleeding in the lungs was observed.

Experimental results: In a mouse pulmonary hemorrhage model, with ibrutinib as a control, the exemplary compound can effectively improve the symptoms of pulmonary hemorrhage, and the lung weight and spleen weight are also significantly lower than the control combination ibrutinib group.

In the description of this specification, descriptions with reference to the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" etc. mean specific features described in conjunction with the embodiment or example , Structure, materials or features are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the described specific features, structures, materials or characteristics may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine the different embodiments or examples and the features of the different embodiments or examples described in this specification without contradicting each other.

Although the embodiments of the present invention have been shown and described above, it can be understood that the above-mentioned embodiments are exemplary and should not be construed as limiting the present invention. A person of ordinary skill in the art can comment on the above-mentioned embodiments within the scope of the present invention. The embodiment undergoes changes, modifications, substitutions, and modifications.

Claims (18)

A compound, which is a compound represented by formula I or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or precursors thereof medicine: X-Y-Z Formula I among them: The Y is

And each E is independently an amide group, an ester group, a carbamate group, a ureido group, a guanidyl group, a heterocyclic group, a cycloalkyl group, a spiro heterobicyclic group, a fused heterobicyclic group, a bridged heterobicyclic group or C6-10 Aryl; each E is optionally substituted by 1, 2, 3 or 4 independent R _b ; each W is O, S, NH or Se, b is an integer between 1-30, and d is 0-30 integer, L _6, L ₇ is independently a bond, -O -, - S (= O) t1 -, - S -, - N (R a) -, - C (= O) O -, - N (R _a )-C(=O)-, -C(=O)-(CH ₂ ) _t2 -, -CH ₂ -, -C(=O)-, -OC(=O)-, -C( =S)-, -C(=O)-N(R _a )-, -C(=S)-N(R _a )-, -(CH ₂ ) _t2 -C(=O)- or triazole base, The X is

The Z is:

Or the Y is

d is an integer between 0 and 30, where, d=6, L ₆ and L _{7 are} independently -N(R _a )-, when -C(=O)-, the X is

The Z is:

d≠6, L ₆ and L _{7 are} independently bonds, -O-, -S(=O) _t1 -, -S-, -N(R _a )-, -C(=O)O-, -N( R _a )-C(=O)-, -CH ₂ -, -C(=O)-, -OC(=O)-, -C(=S)-, -C(=O)-N(R _a )-,-C(=S)-N(R _a )- or triazolyl, the X is

The Z is:

Or the Y is

And W is CH ₂ , O, S, NH or Se, b is an integer between 0 and 30, and d is an integer between 0 and 30, The X is

The Z is:

Or the Y is

And W is CH ₂ , O, S, NH or Se, b is an integer between 0 and 30, and d is an integer between 0 and 30, The X is

When, the Z is:

The X is

时，The Z is

Wherein, each of L ₁ , L ₂ , and L _{3 is} independently a bond, -O-, -S(=O) _t1 -, -S-, -N(R _a )-, -C(=O)O-,- N(R _a )-C(=O)-, -C(=O)-(CH ₂ ) _t2 -, -CH ₂ -, -C(=O)-, -OC(=O)-, -C (=S)-, -C(=O)-N(R _a )-, -C(=S)-N(R _a )-, -(CH ₂ ) _t2 -C(=O)- or trinitrogen Azole Each R _a is independently hydrogen, C1-4 alkyl, halo C1-4 alkyl, C1-4 alkyl group or a hydroxyl group; Each t2 is independently 0, 1, 2, 3 or 4; Each t1 is independently 0, 1 or 2; Each of X ₁ , X ₂ , X ₃ , X ₄ , X ₅ , X ₆ , X ₇ , X ₈ , and Q is independently CH, CH ₂ , O, S, N, NH or Se; Each R ₁ , R ₂ , R ₃ , R ₄ , R _{5 is} independently H, deuterium, amino, C1-4 amide, C1-4 alkyl, C1-4 heteroalkyl, C3-8 cycloalkyl, C2-10 heterocyclyl, C6-10 aryl, C1-9 heteroaryl, C6-10 aryl, C1-4 alkoxy, C1-4 alkenyl, C1-4 alkynyl, wherein, the C1-4 alkyl, C1-4 heteroalkyl, C3-8 cycloalkyl, C2-10 heterocyclyl, C6-10 aryl, C1-9 heteroaryl, C6-10 aryl, C1-4 alkane The oxy, C1-4 alkenyl, and C1-4 alkynyl groups can be optionally selected by one or more selected from deuterium, hydroxyl, amino, oxo, F, Cl, Br, I, cyano, C1-6 alkyl , C1-6 haloalkyl, C1-6 hydroxyalkyl, C1-6 aminoalkyl, C1-6 alkoxy C1-6 alkyl, C1-6 alkylamino C1-6 alkyl, C6-10 aryl C1 -6 alkyl, C1-9 heteroaryl C1-6 alkyl, C2-10 heterocyclyl C1-6 alkyl, C3-10 cycloalkyl C1-6 alkyl, C1-6 alkoxy, C1- 6Alkylamino, C6-10 aryl, C6-10 aryloxy, C6-10 arylamino, C1-9 heteroaryl, C1-9 heteroaryloxy, C2-6 alkenyl, C3-10 cycloalkyl , Halogen-substituted C6-10 aryl, halogen-substituted C1-9 heteroaryl, halogen-substituted C2-10 heterocyclic group or C2-10 heterocyclic group substituted; Each A is independently hydrogen, C1-4 alkyl, C1-4 haloalkyl, hydroxy, nitro, amino, cyano, halogen, carboxy, C1-4 alkoxy, C1-4 alkylamino, C1-4 alkylthio Group, C1-4 alkyl acyl group, C3-12 cycloalkyl group, optionally substituted C3-9 heterocyclic group, optionally substituted C6-12 aryl group, optionally substituted C1-9 heteroaryl group; Each R _{b is} independently hydrogen, C1-4 alkyl, C1-4 haloalkyl, hydroxy, nitro, amino, cyano, halogen, carboxy, C1-4 alkoxy, C1-4 alkylamino, C1-4 Alkylthio, C1-4 alkyl acyl, C3-12 cycloalkyl, C3-9 heterocyclyl, C6-12 aryl, C1-9 heteroaryl, amino C1-4 alkyl, hydroxy C1-4 alkane Group, sulfonic acid group, aminosulfonyl group or aminoacyl group; Each of a, c, e, and f is independently an integer between 0-30. The compound of claim 1, wherein: Each L _{1 is} independently a bond, -O-, -S-, -NH-, Each L _{2 is} independently a bond, -(C=O)CH=CH ₂ -, -O-, -S-, -S(=O)-, -S(=O) ₂ -, -NRa-,- C(=O)-, -C(=O)O-, -N(R _a )-C(=O)-, -CH ₂ -, -OC(=O)-, -C(=S)- , -C(=O)-N(R _a )-, -C(=S)-N(R _a ) or triazolyl, Each R _a is independently hydrogen, C1-2 alkyl, halo C1-2 alkyl, C1-2 alkyl group or a hydroxyl group, Each R ₁ , R ₂ , R ₃ , R _{4 is} independently H, amino, C1-4 alkyl, C1-4 heteroalkyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aromatic Group, C5-7 heteroaryl, wherein the C1-4 alkyl, C1-4 heteroalkyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aryl, C5-7 hetero The aryl group may be optionally substituted with one or more substituents selected from deuterium, hydroxyl, amino, oxo, F, Cl, Br, I, cyano. The compound according to claim 1, wherein each X is independently a compound as shown below:

The compound of claim 1, wherein: Each A is independently hydrogen, optionally substituted C5-7 heterocyclyl, C5-7 cycloalkyl, optionally substituted C6-7 aryl, optionally substituted C5-7 heteroaryl, Each R _{5 is} independently H, amino, C1-4 alkyl, C1-4 heteroalkyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aryl, C5-7 heteroaryl, Wherein, the C1-4 alkyl group, C1-4 heteroalkyl group, C5-7 cycloalkyl group, C5-7 heterocyclic group, C6-7 aryl group, C5-7 heteroaryl group may be optionally substituted by one or Multiple selected from deuterium, hydroxyl, amino, oxo, F, Cl, Br, I, cyano, C5-7 aryl, C5-7 heteroaryl, halogen substituted C5-7 aryl, halogen substituted C5-7 Heteroaryl, halogen-substituted C5-7 heterocyclyl substituents; Each L _{3 is} independently a bond, -(C=O)CH=CH ₂ -, -O-, -S-, -S(=O)-, -S(=O) ₂ -, -NRa-,- C(=O)-, -C(=O)O-, -N(R _a )-C(=O)-, -CH ₂ -, -OC(=O)-, -C(=S)- , -C(=O)-N(R _a )-, -C(=S)-N(R _a ) or triazolyl, Each R _a is independently hydrogen, C1-2 alkyl, halo C1-2 alkyl, C1-2 alkyl group or a hydroxyl group. The compound of claim 1, wherein each A is independently

Each R _{c is} independently hydrogen or C1-2 alkyl. The compound according to claim 1, wherein each Z is independently a compound as shown below:

The compound of claim 1, wherein: Each E is independently an amide group, an ester group, a carbamate group, a ureido group, a guanidino group, a heterocyclic group, a cycloalkyl group, or a C6-C8 aryl group; each E is optionally independently 1, 2, 3, or 4 Replaced by R _b, Each R _{b is} independently hydrogen, C1-2 alkyl, C1-2 haloalkyl, hydroxy, nitro, amino, cyano, halogen, carboxy, C1-2 alkoxy, C1-2 alkylamino, C1-2 Alkylthio, C1-2 alkyl acyl, C5-7 cycloalkyl, C5-7 heterocyclyl, C6-7 aryl, C5-7 heteroaryl, amino C1-2 alkyl, hydroxy C1-2 alkane Group, sulfonic acid group, aminosulfonyl group or aminoacyl group, Each R _a is independently hydrogen, C1-2 alkyl, halo C1-2 alkyl, C1-2 alkyl group or a hydroxyl group. The compound of claim 1, wherein each Y is independently

The compound of claim 1, wherein the X is

Optionally, the Z is

Optionally, the Y is

Wherein, m1 is an integer between 1-10, and n1 is an integer between 0-10. The compound according to any one of claims 1-9, characterized in that it comprises a compound represented by any one of formulas (1) to (15) or its stereoisomers, geometric isomers, and tautomers , Nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs,

Wherein, m1 is an integer between 1-10, and n1 is an integer between 0-10. A compound characterized in that it is a compound represented by any one of formulas 1-18 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, and metabolites , Pharmaceutically acceptable salts or prodrugs,

A pharmaceutical composition characterized by comprising the compound of any one of claims 1-11. The pharmaceutical composition according to claim 12, further comprising a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof. The pharmaceutical composition according to claim 12, further comprising other drugs for the treatment or prevention of non-Hodgkin's lymphoma or autoimmune diseases; Optionally, the autoimmune disease is arthritis, pulmonary hemorrhage, systemic lupus erythematosus, gastrocytosis, chronic lymphatic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, chronic ulcerative colitis, Pernicious anemia with chronic atrophic gastritis, primary biliary cirrhosis, multiple cerebrospinal sclerosis or acute idiopathic polyneuritis; Optionally, the autoimmune disease is arthritis or pulmonary hemorrhage; Optionally, the other drugs for treating or preventing non-Hodgkin's lymphoma or autoimmune diseases include ibrutinib. Use of the compound according to any one of claims 1-11 or the pharmaceutical composition according to any one of claims 12 to 14 in the preparation of a medicine, which is used to degrade BTK or inhibit BTK. The use of the compound according to any one of claims 1-11 or the pharmaceutical composition according to any one of claims 12-14 in the preparation of a medicament for the treatment or prevention of BTK-related diseases. The use according to claim 16, wherein the BTK-related disease is non-Hodgkin's lymphoma or an autoimmune disease. The use according to claim 17, wherein the autoimmune disease is arthritis, pulmonary hemorrhage, systemic lupus erythematosus, corpus sores, chronic lymphatic thyroiditis, hyperthyroidism, insulin-dependent diabetes, severe disease Myasthenia, chronic ulcerative colitis, pernicious anemia with chronic atrophic gastritis, primary biliary cirrhosis, multiple cerebrospinal sclerosis or acute idiopathic polyneuritis; Optionally, the autoimmune disease is arthritis or pulmonary hemorrhage.

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