CN109721597B - Pyridoazepine derivative, and pharmaceutical composition and application thereof - Google Patents

Abstract

The invention discloses a pyrido-azepine derivative, a pharmaceutical composition thereof and application thereof. The pyridoazepine derivative (I), its isomer, prodrug, stable isotope derivative or pharmaceutically acceptable salt thereof has the following structure. The pyridoazepine derivative has good regulation effect on a TLR family and a related signal path, especially has good regulation effect on TLR8, can effectively treat, relieve and/or prevent various diseases mediated by the TLR family and the TLR-related signal path, and particularly can effectively treat, relieve and/or prevent various diseases mediated by TLR8, such as cancer, autoimmune diseases, infection, inflammation, transplant rejection, graft-versus-host disease and the like.

Description

Pyridoazepine derivative, and pharmaceutical composition and application thereof

Technical Field

The invention relates to a pyridoazepine derivative, a pharmaceutical composition and application thereof.

Background

The Toll-like receptor family (TLRs) are important protein families for recognizing pathogen-associated molecular patterns, can sense and initiate innate immune responses and promote the development of adaptive immune responses. TLRs are mainly expressed in immune cells such as myeloid dendritic cells (mDC), plasmacytoid dendritic cells (pDC), monocytes and B cells (Kawai and Akira,2010) and lungs. In humans, over 10 TLRs are considered to have significant functions. TLRs 1/2/4/5 and 6 are located in the cell membrane and function primarily to recognize extracellular macromolecular ligands from bacteria and fungi. In contrast, TLR3/7/8/9 is located in the endosomal membrane within the cell and functions primarily to recognize exogenous nucleic acids from within the cell of the pathogen. Although most TLRs act through specific signaling pathways (primarily through MyD 88-dependent pathways), different TLRs may coordinate different downstream molecules. The addition of specific TLRs leads to the activation of different cell populations (schreibel, et al,2010) and the production of different patterns of cytokines and other inflammatory mediators (Ghosh, et al,2006), leading to different immune responses. For example, upon ligand binding, TLR8 dimerizes and undergoes a conformational change leading to the involvement of the adaptor protein, MyD88, and MyD88 recruits interleukin 1 receptor-associated kinases leading to the activation of downstream signaling pathways including the mitogen-related protein kinases and the transcription factor NF- κ B.

Endosomal TLRs, mainly TLR7/8/9, have been considered as highly attractive new targets for anticancer immunotherapy (Kanzler, et al, 2007; Kreig 2008; Smits, et al, 2008; Hennessy, et al, 2010; Kaczanowska, et al, 2013; Beesu, et al, 2016). For example, TLR7 activates pDCs in response to viral infection, induces high levels of interferon alpha, and induces adaptive T cell responses of major cells to endogenous viral antigens (Liu, et al, 2009). Compared with TLR7/9, TLR8 is more widely expressed in different subtypes of immune cells. Regulatory T cells (tregs) have potent immune response suppressive capacity and are a major obstacle to effective cancer immunotherapy. The TLR8 signaling pathway was demonstrated to be a necessary and sufficient condition to reverse Treg cell suppression function leading to strong tumor suppression. TLR8 selective agonists are effective in activating a variety of immune cells, including mDCs and monocytes (Gorden, et al,2005), and can promote the generation of an adaptive immune response against cancer cellsRaw (Krug, et al, 2003; Schnurr, et al, 2005). Activated mDCs phagocytose apoptotic and dead tumor cells, which in turn, are more potent towards CD8 than pDCs⁺CTLs cross-present tumor-associated antigens (Berard, et al, 2000; Dalgaard, et al, 2005). In addition, mDCs are activated, resulting in the release of TNF α and interleukin 12(IL-12) which stimulate the activation of T cells and NK cells. NK cell activation is the primary mechanism of antibody-mediated cytotoxicity (ADCC). Thus, potentiation of tumor cell killing by ADCC may present an important therapeutic opportunity for selective inhibitors of TLR8 (Lu, et al, 2011). Some monoclonal antibody therapies are widely used for the treatment of cancer patients, such as rituximab and trastuzumab, which can play a therapeutic role through ADCC (Ferris, et al, 2010). Indeed, the addition of TLR8 agonists in mAb therapy can enhance ADCC and thus increase the efficacy of mAb therapy (Ferris, et al, 2015). In addition, recent studies have shown that TLR8 agonists can directly act as anti-tumor agents, independent of its immunomodulatory function (Ignatz-Hoover, et al, 2015). Thus, TLR8 agonists not only act as monotherapies, but also enhance the efficacy of various chemotherapeutic and targeted anticancer drugs by enhancing host immune responses.

Among the members of the TLRs family which recognize pathogenic microorganism nucleic acids, TLR7 and TLR8 have high homology, and can recognize some artificially synthesized small molecules with antiviral effects, such as Imidazoquinolines imidazoquinoline small molecule compounds (ligands of TLR7 and TLR 8). Imidazoquinolines were studied in a model of genital herpes in guinea pigs infected with HSV and found to be less effective against viral replication in vitro but more effective in vivo, suggesting that this class of compounds promotes the production of pro-inflammatory factors by immune cells and modulates cytokines, leading to antiviral responses (Int Immunopharmacol 2002; 2: 443-451). More importantly, TLR7 and TLR8 recognize viral ssRNA. Research has shown that ssRNA viruses are natural ligands for TLR7 and TLR8, such as human immunodeficiency virus type I (HIV), influenza, Sendai virus, dengue virus, Newcastle Disease Virus (NDV), Vesicular Stomatitis Virus (VSV), and Hepatitis C Virus (HCV). The TLR8 can identify antiviral compounds, ssRNA viruses, artificially synthesized oligonucleotides and the like, induces Th1 through a MyD88 dependent signal channel, inhibits Th2 cytokine secretion and Tregs proliferation, mediates antiviral immunity, and plays anti-infection and anti-allergic effects.

The X-ray crystal form structure of TLR8 protein and its associated TLR family members have been extensively studied, which will further facilitate structure-based drug design and optimization.

Thus, TLR8 is currently an attractive therapeutic target. Despite the much research on TLRs, there is still a great opportunity to further expand their use and advantages. The compounds and uses described in the present invention will contribute to the development of TLR8 agonists, meeting clinically unmet needs.

Disclosure of Invention

The invention aims to solve the technical problem of providing a novel pyridoazepine derivative, a pharmaceutical composition thereof and application thereof. The pyridoazepine derivative has a good regulation effect on TLR8, and can effectively treat, relieve and/or prevent various related diseases caused by immunosuppression, such as cancer, autoimmune diseases, infection, inflammation, transplant rejection, graft-versus-host disease and the like.

The invention provides a compound shown as a formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof;

wherein X is N or CR₄(ii) a Y is N or CR₄(ii) a And X and Y are not CR at the same time₄；

L is a connecting bond, C_2-6Alkenylene radical, C_2-6Alkynylene, or- (CH)₂)_m-；

R is a benzene ring, a 5-6 membered heteroaromatic ring, or a fused cyclic group, said R being unsubstituted or optionally substituted with one or more of R₅Substitution at any position: hydrogen, halogen, alkyl, haloalkyl, haloalkoxy, alkenyl, alkynyl, Cy¹、-L₁-Cy¹、-CN、-NO₂、-SR_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(O)NR_dS(O)₂R_e、-C(＝NH)R_e、-C(＝NH)NR_dR_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dS(O)₂R_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-S(O)_1-2R_e、-S(O)₂NR_dR_eand-NR_dS(O)₂NR_dR_e(ii) a Wherein the alkyl, alkenyl or alkynyl is unsubstituted or optionally substituted at any position with one or more of the following substituents: -CN, -NO₂、-SR_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(O)NR_dS(O)₂R_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-NR_dS(O)₂R_e、-NR_dS(O)₂NR_dR_e、-N(R_d)C(O)N(R_d)S(O)₂R_e、-S(O)_1-2R_e、-S(O)₂NR_dR_e、-S(O)(＝NCN)R_e、-S(O)(＝NR_d)R_e、-S(O)(＝NSO₂R_d)R_e、-S(O)₂N(R_d)C(O)NR_dR_e、-P(O)(OR_d)₂、-OP(O)(OR_d)₂、-S(O)₂N(R_d)C(O)R_eand-B (OR)_d)₂；

L₁Is a connecting bond, C_2-6Alkenylene radical, C_2-6Alkynylene, or- (CR)_aR_b)_n-；

Cy¹Is cycloalkyl, heterocycloalkyl, aryl or heteroaryl; the Cy is¹Is unsubstituted or optionally substituted in any position with one or more of the following groups: halogen, alkyl, haloalkyl, haloalkoxy, hydroxyalkyl, aminoalkyl, alkenyl, alkynyl, -CN, -NO₂、-SR_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dS(O)₂R_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-S(O)_1-2R_e、-S(O)₂NR_dR_eand-NR_dS(O)₂NR_dR_e；

R₁And R₂Each independently selected from hydrogen or C_1-6An alkyl group;

or, R₁And R₂Together with the N atom to which they are both attached form a 3-8 membered heterocycloalkyl group; the heterocycloalkyl group may further contain 1 to 3 hetero atoms selected from N, O, S;

R₃and R₄Each independently selected from hydrogen, deuterium, halogen, amino, cyano, nitro, C_1-6Alkyl, halo C_1-6Alkyl, halo C_1-6Alkoxy radical, C_1-6Alkoxy radical, C_2-6Alkenyl radical, C_2-6Alkynyl, or hydroxy C_1-6An alkyl group;

R_aand R_bEach independently selected from hydrogen, deuterium, halogen or C_1-6An alkyl group;

or, R_aAnd R_bC atom co-linked with themTogether form C_3-8Cycloalkyl or 3-8 membered heterocycloalkyl;

each R_dAnd each R_eEach independently selected from hydrogen and C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_3-8Cycloalkyl, 3-8 membered heterocycloalkyl, C_6-10Aryl, 5-6 membered heteroaryl, C_3-8Cycloalkyl radical C_1-6Alkyl, 3-8 membered heterocycloalkyl C_1-6Alkyl, phenyl C_1-6Alkyl, 5-6 membered heteroaryl C_1-6An alkyl group; the R is_dOr R_eIs unsubstituted or optionally substituted by 1 to 3 substituents selected from halogen, hydroxy, amino, carboxy, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Alkylamino, halogeno C_1-6Alkyl, halo C_1-6Alkoxy radical, C_2-6Alkenyl radical, C_2-6One or more substituents in the alkynyl group are substituted at any position;

or, R_dAnd R_eTogether with the N atom to which they are both attached form a 3-8 membered heterocycloalkyl group; the heterocycloalkyl group may further contain 1 to 3 heteroatoms selected from N, O, S; the heterocycloalkyl is unsubstituted or further substituted by 1 to 3 groups selected from halogen, amino, hydroxy, carboxy, cyano, C_1-6Alkyl, halo C_1-6Alkyl radical, C_2-6Alkenyl radical, C_2-6Alkynyl, C_1-3Alkyl, amino C_1-3Alkyl, -OR_d1、-OC(O)R_d1、-OC(O)OR_d1、-OC(O)NR_d1R_e1、-C(O)OR_d1、-C(O)R_d1、-C(O)NR_d1R_e1、-NR_d1R_e1、-NR_d1C(O)R_e1、-N(R_d1)C(O)OR_e1、-N(R_d1)C(O)NR_d1R_e1、-NR_d1S(O)₂R_e1、-NR_d1C(＝NH)R_e1、-NR_d2C(＝NH)NR_d1R_e1、-NR_d1S(O)₂NR_d1R_e1、-S(O)_1-2R_d1and-S (O)₂NR_d1R_e1Is substituted at any position with one or more substituents of (a);

R_d1and R_e1Each independently selected from hydrogen or C_1-6An alkyl group;

m or n is an integer of 1 to 6.

The L is preferably a bond.

The R is₁Preferably n-propyl.

The R is₂Preferably n-propyl.

The R is₃Preferably H, D, F, Cl, Br, -CN, -NH₂、-CH₃、-OCH₃、-OCF₃、-CH₂F、-CHF₂or-CF₃

The R is₄Preferably H.

The R is preferably an 8-to 20-membered bicyclic or tricyclic fused ring group.

More preferably, R is a 9-to 15-membered bicyclic or tricyclic fused ring group, and at least one ring is an aromatic ring.

In some preferred embodiments, R is a 9-15 membered bicyclic or tricyclic fused ring group, one of the rings is aromatic and the other 1-2 rings are non-aromatic, the 9-15 membered bicyclic or tricyclic fused ring group includes 1-3N atoms in the ring atoms, and the non-aromatic ring further includes 1-2 oxo groups

And/or thio groups

The R is unsubstituted or preferably 1 to 4R₅Substitution at any position; more preferably 1,1 to 2, or 1 to 3R₅The substitution is in any position.

More preferably, R is of the structure:

wherein R is₅Is hydrogen, halogen, alkyl, haloalkyl,Haloalkoxy, alkenyl, alkynyl, Cy¹、-L₁-Cy¹、-SR_d、-OR_d、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(O)NR_dS(O)₂R_e、-C(＝NH)R_e、-C(＝NH)NR_dR_e、-S(O)₂R_eand-S (O)₂NR_dR_e(ii) a Wherein the alkyl, alkenyl or alkynyl is unsubstituted or optionally substituted at any position with 1 to 3 substituents as follows: -CN, -NO₂、-SR_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(O)NR_dS(O)₂R_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-NR_dS(O)₂R_e、-NR_dS(O)₂NR_dR_e、-N(R_d)C(O)N(R_d)S(O)₂R_e、-S(O)_1-2R_e、-S(O)₂NR_dR_e、-S(O)(＝NCN)R_e、-S(O)(＝NR_d)R_e、-S(O)(＝NSO₂R_d)R_e、-S(O)₂N(R_d)C(O)R_e、-S(O)₂N(R_d)C(O)NR_dR_e、-P(O)(OR_d)₂、-OP(O)(OR_d)₂OR-B (OR)_d)₂；L₁、Cy¹、R_dAnd R_eAs defined above.

The R is₅The halogen is preferably F, Cl, or Br.

The R is₅In (1), the alkyl group is preferably C_1-6Alkyl groups, for example: methyl, ethyl, isopropyl, n-butyl, isobutyl, tert-butyl, neopentyl, tert-pentyl, or 2, 2-dimethylbutyl。

The R is₅The haloalkyl group is preferably a halogeno C_1-6Alkyl groups, for example: -CF₃、-CHF₂or-CH₂F。

The R is₅The haloalkoxy group is preferably a halogeno group C_1-6Alkoxy groups, for example: -OCF₃、-OCHF₂or-OCH₂F。

The R is₅In (1), the alkenyl group is preferably C_2-6An alkenyl group.

The R is₅In (1), the alkynyl group is preferably C_2-6Alkynyl.

The R is₅The alkyl group, alkenyl group or alkynyl group is unsubstituted or substituted at any position with preferably 1 to 3 substituents.

The R is₅In (a), the-L₁-Cy¹In (1), the L₁Preferably a connecting bond, -CH₂-、-CH₂CH₂-、-CH₂CH₂CH₂-、-CH₂CH₂C(CH₃)₂-、-CH₂CH₂CH₂CH₂-, or-CH₂CH(CH₃)CH₂-；

The R is₅In (a), the-L₁-Cy¹In (1), the Cy¹Preferably C_3-10Cycloalkyl, 3-10 membered heterocycloalkyl, C_6-10Aryl, or 5-10 membered heteroaryl.

The R is₅In (a), the-L₁-Cy¹In (1), the Cy¹More preferably, the 5-to 10-membered heteroaryl group is a 5-6-membered heteroaryl group such as: pyridyl, pyrimidinyl, piperazinyl, pyrazinyl, furanyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, 1,2, 5-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,3, 4-thiadiazole, 1,2, 4-triazolyl, 1,2, 3-triazolyl, tetrazolyl;

the R is₅In (1), the Cy¹Is unsubstituted or optionally substituted by 1 to 3 substituents selected from F, Cl, Br, C_1-4Alkyl, halo C_1-3Alkyl and halo C_1-3One or more substituents in the alkoxy group are substituted at any position.

The R is₅More preferably C_1-6Alkyl, or-L₁-Cy¹(ii) a Wherein said C_1-6The alkyl group is unsubstituted or optionally substituted at any position with 1 to 3 substituents as follows: -SR_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dS(O)₂R_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-S(O)_1-2R_e、-S(O)₂NR_dR_e、-S(O)(＝NCN)R_e、-S(O)(＝NR_d)R_e、-S(O)(＝NSO₂R_d)R_eor-NR_dS(O)₂NR_dR_e；R_dAnd R_eAre each independently hydrogen or C_1-6An alkyl group.

The R is₅More preferably a propyl group, an isopropyl group, a butyl group, an isobutyl group, a n-butyl group, a neopentyl group, a tert-pentyl group, a,

The compound shown in the formula I, the isomer, the prodrug, the stable isotope derivative or the pharmaceutically acceptable salt thereof is preferably the compound shown in the formula II, the isomer, the prodrug, the stable isotope derivative or the pharmaceutically acceptable salt thereof:

wherein, U is CH or N; v is O or S; x is N and Y is CH, or X is CH and Y is N;

R₅the definitions of (A) and (B) are as described above.

The compound shown in the formula I, the isomer, the prodrug, the stable isotope derivative or the pharmaceutically acceptable salt thereof is optimally any one of the following structures:

the compound shown as the formula (I) and pharmaceutically acceptable salts thereof can be synthesized by a general chemical method.

In general, salts can be prepared by reacting the free base or acid with equal chemical equivalents or an excess of acid (inorganic or organic) or base (inorganic or organic) in a suitable solvent or solvent composition.

The invention also provides a pharmaceutical composition, which comprises a therapeutically effective amount of active components and pharmaceutically acceptable auxiliary materials; the active component comprises one or more of a compound shown as a formula (I), an isomer, a prodrug, a stable isotope derivative and a pharmaceutically acceptable salt thereof.

In the pharmaceutical composition, the active ingredient may also include other therapeutic agents for cancer, viral infections or autoimmune diseases.

In the pharmaceutical composition, the pharmaceutically acceptable adjuvant may include a pharmaceutically acceptable carrier, diluent and/or excipient.

The pharmaceutical composition may be formulated into various types of administration unit dosage forms, such as tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, suppositories, injections (solutions and suspensions), and the like, preferably liquids, suspensions, emulsions, suppositories, injections (solutions and suspensions), and the like, according to the therapeutic purpose.

For shaping the pharmaceutical composition in the form of tablets, any excipient known and widely used in the art may be used. For example, carriers such as lactose, white sugar, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, and the like; binders such as water, ethanol, propanol, common syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, shellac, methyl cellulose and potassium phosphate, polyvinylpyrrolidone, etc.; disintegrators such as dry starch, sodium alginate, agar powder and kelp powder, sodium bicarbonate, calcium carbonate, fatty acid esters of polyethylene sorbitan, sodium lauryl sulfate, monoglyceride stearate, starch, lactose and the like; disintegration inhibitors such as white sugar, glycerol tristearate, coconut oil and hydrogenated oil; adsorption promoters such as quaternary ammonium bases and sodium lauryl sulfate, etc.; humectants such as glycerin, starch, and the like; adsorbents such as starch, lactose, kaolin, bentonite, colloidal silicic acid, and the like; and lubricants such as pure talc, stearates, boric acid powder, polyethylene glycol, and the like. Optionally, conventional coating materials can be selected to make into sugar-coated tablet, gelatin film-coated tablet, enteric coated tablet, film-coated tablet, double-layer film tablet and multilayer tablet.

For shaping the pharmaceutical composition in the form of pellets, any of the excipients known and widely used in the art may be used, for example, carriers such as lactose, starch, coconut oil, hardened vegetable oil, kaolin, talc and the like; binders such as gum arabic powder, tragacanth powder, gelatin, ethanol and the like; disintegrating agents, such as agar and kelp powder.

For shaping the pharmaceutical composition in the form of suppositories, any excipient known and widely used in the art may be used, for example, polyethylene glycol, coconut oil, higher alcohols, esters of higher alcohols, gelatin, semisynthetic glycerides and the like.

For preparing the pharmaceutical composition in the form of injection, the solution or suspension may be sterilized (preferably by adding appropriate amount of sodium chloride, glucose or glycerol) and made into injection with blood isotonic pressure. In the preparation of injection, any carrier commonly used in the art may also be used. For example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, and fatty acid esters of polyethylene sorbitan, and the like. In addition, conventional lytic agents, buffers, analgesics, and the like may be added.

In the present invention, the content of the composition in the pharmaceutical composition is not particularly limited, and can be selected from a wide range, and generally ranges from 5 to 95% by mass, preferably from 30 to 80% by mass.

In the present invention, the method of administration of the pharmaceutical composition is not particularly limited. The formulation of various dosage forms can be selected for administration according to the age, sex and other conditions and symptoms of the patient. For example, tablets, pills, solutions, suspensions, emulsions, granules or capsules are administered orally; the injection can be administered alone or mixed with infusion solution (such as glucose solution and amino acid solution) for intravenous injection; the suppository is administered to the rectum.

The invention also provides a compound shown as the formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, or an application of the pharmaceutical composition in preparing TLRs regulators. Such modulators of TLRs include agonists of TLRs, or partial agonists of TLRs. Preferably, the TLRs are TLR 8.

The invention also provides a compound shown as the formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, or an application of the pharmaceutical composition in preparing a medicine for regulating T cells and other immune cells.

The invention also provides an application of the compound shown as the formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition in preparing a medicament for treating, relieving and/or preventing related diseases mediated by TLR 8; the invention preferably provides an application of the compound shown as the formula (I), an isomer, a prodrug, a stable isotope derivative or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition in preparing a medicament for treating, relieving and/or preventing related diseases mediated by TLR 8; such diseases include neoplastic and non-neoplastic diseases. Such diseases include, but are not limited to: cancer, viral and other infections, diseases due to immunosuppression, and autoimmune diseases, among others.

The invention preferably uses the compound shown in the formula (I), isomers, prodrugs, stable isotope derivatives or pharmaceutically acceptable salts thereof, or the application of the pharmaceutical composition in preparing medicines for treating and/or relieving cancers. The cancer is preferably an immune agent-associated cancer, and the immunosuppression refers to tumor-specific immunosuppression.

The present invention still further provides methods of treating cancer, viral and other infections, autoimmune diseases with said compounds of formula (I), isomers, prodrugs, stable isotopic derivatives or pharmaceutically acceptable salts thereof, or said pharmaceutical compositions comprising: administering to a mammal a therapeutically desired dose of a compound according to formula (I), an isomer, prodrug, stable isotopic derivative or pharmaceutically acceptable salt thereof, or a pharmaceutical composition.

The mammal, preferably a human.

The invention further provides the compounds as shown in the formula (I), isomers, prodrugs, stable isotope derivatives or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions can be combined with one or more other kinds of therapeutic agents and/or treatment methods to treat, alleviate and/or prevent related diseases mediated by TLRs, particularly related diseases mediated by TLR 8. The related diseases mediated by the TLR8 refer to diseases caused by TLR 8-mediated immunosuppression, and the diseases can comprise: cancer, viral and other infections, inflammation, autoimmune diseases, transplant rejection, graft versus host disease, and the like.

The compound shown in the formula (I), isomers, prodrugs, stable isotope derivatives or pharmaceutically acceptable salts thereof or the pharmaceutical composition can be combined with one or more other kinds of therapeutic agents and/or therapeutic methods to treat and/or relieve diseases mediated by tryptophan TLR8, wherein the diseases are preferably cancer.

The other therapeutic agent (e.g. other therapeutic agents useful in the treatment of cancer) may be administered in a single therapeutic form or in separate therapeutic forms in succession with the compound of formula (I).

The viral infection may include: infection by viruses such as influenza virus, sendai virus, coxsackie virus, dengue virus, Newcastle Disease Virus (NDV), Vesicular Stomatitis Virus (VSV) bab, Hepatitis C Virus (HCV), Hepatitis B Virus (HBV), Human Papilloma Virus (HPV), Cytomegalovirus (CMV), epstein-barr virus (EBV), poliovirus, varicella-zoster virus, or human immunodeficiency virus type I (HIV).

The cancer includes metastatic and non-metastatic cancer, also includes familial hereditary and sporadic cancer, and also includes solid tumor and non-solid tumor.

Specific examples of the solid tumor may include, but are not limited to: one or more of eye cancer, bone cancer, lung cancer, stomach cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer (including glioblastoma, medulloblastoma), ovarian cancer, bladder cancer, cervical cancer, testicular cancer, kidney cancer (including adenocarcinoma and nephroblastoma), oral cancer (including squamous cell carcinoma), tongue cancer, laryngeal cancer, nasopharyngeal cancer, head and neck cancer, colon cancer, small intestine cancer, rectal cancer, parathyroid cancer, thyroid cancer, esophageal cancer, gallbladder cancer, bile duct cancer, cervical cancer, liver cancer, lung cancer, sarcoma, and skin cancer.

Specific examples of such non-solid tumors (including hematological tumors) may include, but are not limited to: one or more of lymphoid leukemia (including acute lymphocytic leukemia, lymphoma, myeloma, chronic lymphocytic leukemia, hodgkin lymphoma, non-hodgkin lymphoma, T-cell chronic lymphocytic leukemia, B-cell chronic lymphocytic leukemia), myeloid related leukemia (including acute myeloid leukemia, chronic myeloid leukemia), and AIDs-related leukemia.

The autoimmune disease may include, but is not limited to: rheumatoid arthritis, systemic lupus erythematosus, Mixed Connective Tissue Disease (MCTD), systemic scleroderma (including CREST syndrome), dermatomyositis, nodular vasculitis, nephropathy (including hemorrhagic nephritis syndrome, acute glomerulonephritis, primary membranous proliferative glomerulonephritis, etc.), endocrine-related diseases (including type I diabetes, gonadal insufficiency, oxa-anemia, hyperthyroidism, etc.), liver diseases (including primary biliary cirrhosis, autoimmune cholangitis, autoimmune hepatitis, primary sclerosing cholangitis, etc.), and autoimmune reactions due to infection (e.g., AIDS, malaria, etc.).

In the present invention, unless otherwise specified, the term "optionally substituted at any position by one or more groups" means that any one or more hydrogen atoms of one or more atoms specified on the group are substituted with the specified group, provided that the normal valency of the specified atom is not exceeded, said substitution being at any position as is common in the art as reasonable.

In the present invention, unless otherwise specified, chemical bonds show chemical structures represented by dotted lines, meaning that the bonds are optionally present or absent. For example, a dashed line drawn next to a solid single bond indicates that the bond may be a single or double bond.

In the present invention, when the bond to a substituent exhibits an intersection with a bond linking two atoms in the ring, then such substituent may be bonded to any bondable ring atom on the ring.

Unless otherwise indicated, the following terms appearing in the specification and claims of the invention have the following meanings:

the term "alkyl" refers to a saturated straight or branched chain hydrocarbon group containing 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 8, 1 to 6,1 to 4, 1 to 3 carbon atoms, representative examples of alkyl groups including but not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, isobutyl, tert-butyl, tert-,Pentyl, hexyl, heptyl, octyl, nonyl, decyl, 4-dimethylpentyl, 2, 4-trimethylpentyl, undecyl, dodecyl, and their various isomers, and the like. When "alkyl" is used as a linking group for other groups, e.g. - (CH)₂)_m-, which may be branched or unbranched, examples include, but are not limited to-CH₂-、-CH₂CH₂-、-CH₂CH(CH₃)-。

The term "cycloalkyl" refers to a saturated or partially unsaturated (containing 1 or 2 double bonds) monocyclic or polycyclic group containing 3 to 20 carbon atoms. "monocyclic cycloalkyl" is preferably 3-10 membered monocyclic cycloalkyl, more preferably 3-8 membered monocyclic cycloalkyl, for example: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclododecyl, cyclohexenyl. "polycyclic cycloalkyl" includes "bridged cyclic groups", "fused cycloalkyl" and "spirocycloalkyl", representative examples of "bridged cyclic groups" include, but are not limited to: bornyl, bicyclo [2.2.1] heptenyl, bicyclo [3.1.1] heptenyl, bicyclo [2.2.1] heptenyl, bicyclo [2.2.2] octanyl, bicyclo [3.2.2] nonanyl, bicyclo [3.3.1] nonanyl, bicyclo [4.2.1] nonanyl, adamantyl, and the like. "fused cycloalkyl" includes a cycloalkyl ring fused to a phenyl, cycloalkyl, or heteroaryl group, including, but not limited to: benzocyclobutene, 2, 3-dihydro-1-H-indene, 2, 3-cyclopentenopyridine, 5, 6-dihydro-4H-cyclopentyl [ B ] thiophene, decahydronaphthalene and the like. The monocyclic cycloalkyl or polycyclic cycloalkyl groups can be linked to the parent molecule through any carbon atom in the ring.

The term "heterocycloalkyl" refers to a saturated or partially unsaturated (containing 1 or 2 double bonds) non-aromatic cyclic group consisting of carbon atoms and heteroatoms selected from nitrogen, oxygen or sulfur, which may be monocyclic or polycyclic, in the present invention the number of heteroatoms in the heterocycloalkyl is preferably 1,2,3 or 4, and the nitrogen, carbon or sulfur atoms in the heterocycloalkyl may be optionally oxidized. The nitrogen atom may optionally be further substituted with other groups to form tertiary amines or quaternary ammonium salts. "monocyclic heterocycloalkyl" is preferably 3-10 membered monocyclic heterocycloalkyl, more preferably 3-8 membered monocyclic heterocycloalkyl. For example: aziridinyl, tetrahydrofuran-2-yl, morpholin-4-yl, thiomorpholin-S-oxide-4-yl, piperidin-1-yl, N-alkylpiperidin-4-yl, pyrrolidin-1-yl, N-alkylpyrrolidin-2-yl, piperazin-1-yl, 4-alkylpiperazin-1-yl, and the like. "polycyclic heterocycloalkyl" includes "fused heterocycloalkyl", "spiroheterocyclyl" and "bridged heterocycloalkyl". "fused heterocycloalkyl" includes a monocyclic heterocycloalkyl ring fused to a phenyl, cycloalkyl, heterocycloalkyl, or heteroaryl, and fused heterocycloalkyl includes, but is not limited to: 2, 3-dihydrobenzofuranyl, 1, 3-dihydroisobenzofuranyl, indolinyl, 2, 3-dihydrobenzo [ b ] thienyl, dihydrobenzopyranyl, 1,2,3, 4-tetrahydroquinolyl, and the like. Monocyclic heterocycloalkyl and polycyclic heterocycloalkyl can be linked to the parent molecule through any ring atom in the ring. The above ring atoms particularly denote carbon atoms and/or nitrogen atoms constituting the ring skeleton.

The term "cycloalkylalkyl" refers to a cycloalkyl group attached to the parent nuclear structure through an alkyl group. Thus, "cycloalkylalkyl" encompasses the definitions of alkyl and cycloalkyl above.

The term "heterocycloalkylalkyl" refers to a linkage between a heterocycloalkyi and the parent nuclear structure through an alkyl group. Thus, "heterocycloalkylalkyl" embraces the definitions of alkyl and heterocycloalkyl described above.

The term "alkoxy" refers to a cyclic or acyclic alkyl group having the indicated number of carbon atoms attached through an oxygen bridge, including alkyloxy, cycloalkyloxy, and heterocycloalkyloxy. Thus, "alkoxy" encompasses the above definitions of alkyl, heterocycloalkyl, and cycloalkyl.

The term "hydroxyalkyl" means that any one of the hydrogen atoms on the alkyl group is replaced with a hydroxyl group, including, but not limited to: -CH₂OH、-CH₂CH₂OH、-CH₂CH₂C(CH₃)₂OH。

The term "alkenyl" refers to a straight, branched, or cyclic non-aromatic hydrocarbon group containing at least 1 carbon-carbon double bond. Wherein 1-3 carbon-carbon double bonds, preferably 1 carbon-carbon double bond, may be present. The term "C_2-4Alkenyl "means an alkenyl group having 2 to 4 carbon atoms, the term" C_2-6Alkenyl "means alkenyl having 2 to 6 carbon atoms, and includesEthenyl, propenyl, butenyl, 2-methylbutenyl and cyclohexenyl. The alkenyl group may be substituted.

The term "alkynyl" refers to a straight, branched, or cyclic hydrocarbon group containing at least 1 carbon-carbon triple bond. Wherein 1-3 carbon-carbon triple bonds, preferably 1 carbon-carbon triple bond, may be present. The term "C_2-6Alkynyl "refers to alkynyl groups having 2 to 6 carbon atoms and includes ethynyl, propynyl, butynyl, and 3-methylbutynyl.

The term "aryl" refers to any stable 6-20 membered monocyclic or polycyclic aromatic group, for example: phenyl, naphthyl, tetrahydronaphthyl, 2, 3-indanyl, biphenyl, or the like.

The term "heteroaryl" refers to an aromatic ring group formed by replacement of at least 1 ring carbon atom with a heteroatom selected from nitrogen, oxygen or sulfur, which may be a 5-7 membered monocyclic structure or a 7-20 fused ring structure, preferably a 5-6 membered heteroaryl. In the present invention, the number of hetero atoms is preferably 1,2 or 3, and includes pyridyl, pyrimidyl, piperazinyl, pyridazin-3 (2H) -onyl, furyl, thienyl, thiazolyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, 1,2, 5-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,3, 4-thiadiazole, 1,2, 4-triazolyl, 1,2, 3-triazolyl, tetrazolyl, indazolyl, isoindolyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzo [ d ] [1,3] dioxolanyl, benzothiazolyl, benzoxazolyl, quinolyl, isoquinolyl, isoquinolinyl, quinazolinyl, 4-hydroxythieno [3,2-c ] pyridyl, 4, 5-dihydro-4-oxofuro [3,2] pyridyl, 4-hydroxy-5-azaindolyl, furo [2,3-c ] bipyridin-7 (6H) -onyl, thieno [2,3-c ] bipyridin-7 (6H) -onyl, and the like.

The term "fused ring group" refers to a fused ring structure formed by two, three, or four ring structures sharing two adjacent atoms with each other, and each single ring of the ring structures may be a monocyclic aryl, monocyclic heteroaryl, monocyclic cycloalkyl, or monocyclic heterocycloalkyl. The fused ring group referred to in the present invention is a saturated, unsaturated or partially saturated fused ring structure, and preferably at least one ring is an aromatic ring. More preferably a bicyclic or tricyclic fused ring groupAnd at least one ring is aromatic. The non-aromatic ring in the fused ring group may further comprise 1-2 oxo or thioxo groups, preferably 1-2 saturated carbon atoms on the non-aromatic ring are further oxidized to carbonyl groups. In the present invention, the fused cyclic group is an 8-to 20-membered, preferably 8-to 15-membered fused cyclic group. Specific examples of fused cyclic groups include, but are not limited to: benzocyclobutene radical, 2, 3-dihydro-1-H-indenyl, 1,2,3, 4-tetrahydronaphthyl, 6,7,8, 9-tetrahydro-5H-benzo [7 ]]Cycloalkenyl, 6, 9-dihydro-5H-benzo [7 ]]Cycloalkenyl, 5,6,7,8,9, 10-hexahydrobenzo [8 ]]Cycloalkenyl, 2, 3-cyclopentenopyridyl, 5, 6-dihydro-4H-cyclopentyl [ B]Thienyl, 5, 6-dihydro-4H-cyclopentyl [ B]Furyl, 2, 3-dihydrobenzofuryl, 1, 3-dihydroisobenzofuryl, indolinyl, 2, 3-dihydrobenzo [ b ]]Thienyl, dihydrobenzopyranyl, 1,2,3, 4-tetrahydroquinolyl, 2, 3-dihydro-1, 4-benzodioxanyl, 3, 4-dihydro-2H-1, 4-benzoxazinyl, naphthyridinyl, benzofuranyl, benzothienyl, benzopyrolyl, benzothiazolyl, benzoxazolyl, indazolyl, benzopyrazinyl, benzimidazolyl, indolyl, quinolyl, isoquinolyl, purinyl, pteridinyl, quinoxalinyl,

The fused ring group may be attached to the parent molecule through a ring carbon atom, preferably through an aromatic ring carbon atom. The fused cyclic group may be unsubstituted or optionally substituted at any position with one or more substituents.

The term "arylalkyl" refers to an alkyl linkage between an aryl group and the parent nucleus structure. Thus, "arylalkyl" encompasses the above definitions of alkyl and aryl groups.

The term "heteroarylalkyl" refers to a heterocycloalkyl group attached to the parent nucleus structure through an alkyl group. Thus, "heteroarylalkyl" embraces the definitions of alkyl and heteroaryl as described above.

The term "halogen" denotes fluorine, chlorine, bromine or iodine.

The term "haloalkyl" refers to an alkyl group optionally substituted with a halogen. Thus, "haloalkyl" encompasses the above definitions of halogen and alkyl.

The term "haloalkoxy" refers to an alkoxy group optionally substituted with a halogen. Thus, "haloalkoxy" encompasses the above definitions of halogen and alkoxy.

The term "amino" refers to the group-NH₂The term "alkylamino" refers to an amino group wherein at least one hydrogen atom is replaced with an alkyl group, including, but not limited to: -NHCH₃、-N(CH₃)₂、-NHCH₂CH₃、-N(CH₂CH₃)₂. The term "aminoalkyl" refers to an alkyl group wherein any one of the hydrogen atoms is replaced by an amino group, including, but not limited to: -CH₂NH₂、-CH₂CH₂NH₂. Thus, "aminoalkyl" and "alkylamino" encompass the definitions of alkyl and amino as described above.

The term "nitro" means-NO₂。

The term "cyano" refers to — CN.

The symbol "═" represents a double bond; symbol

Represents a double bond or a single bond.

The "room temperature" of the invention means 15-30 ℃.

The isotopically substituted derivatives include: an isotopically substituted derivative in which any hydrogen atom in formula I is substituted with 1 to 5 deuterium atoms, an isotopically substituted derivative in which any carbon atom in formula I is substituted with 1 to 3 carbon 14 atoms, or an isotopically substituted derivative in which any oxygen atom in formula I is substituted with 1 to 3 oxygen 18 atoms.

By "prodrug" is meant a compound that is metabolized in vivo to the original active compound. Prodrugs are typically inactive substances or less active than the active parent compound, but may provide convenient handling, administration, or improved metabolic properties.

The "Pharmaceutically acceptable salts" described herein are discussed in Berge, et al, "pharmaceutical acceptable salts", j.pharm.sci.,66,1-19(1977), and are apparent to the pharmaceutical chemist, as being substantially non-toxic and providing the desired pharmacokinetic properties, palatability, absorption, distribution, metabolism, excretion, etc. The compounds of the present invention may have an acidic group, a basic group or an amphoteric group, and typical pharmaceutically acceptable salts include salts prepared by reacting the compounds of the present invention with an acid, for example: hydrochloride, hydrobromide, sulphate, pyrosulphate, hydrogen sulphate, sulphite, bisulphite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, nitrate, acetate, propionate, decanoate, octanoate, formate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, suberate, benzoate, methylbenzoate, phthalate, maleate, methanesulfonate, p-toluenesulfonate, (D, L) -tartaric acid, citric acid, maleic acid, (D, L) -malic acid, fumaric acid, succinic acid, succinate, lactate, trifluoromethanesulfonate, naphthalene-1-sulfonate, mandelate, pyruvate, stearate, ascorbate, salicylate. When the compound of the present invention contains an acidic group, pharmaceutically acceptable salts thereof may further include: alkali metal salts, such as sodium or potassium salts; alkaline earth metal salts, such as calcium or magnesium salts; examples of the organic base salt include salts with ammonia, alkylamines, hydroxyalkylamines, amino acids (lysine and arginine), and N-methylglucamine.

The term "isomers" as used herein means that the compounds of formula (I) of the present invention may have asymmetric centers and racemates, racemic mixtures and individual diastereomers, and all such isomers, including stereoisomers and geometric isomers, are encompassed by the present invention. In the present invention, when a compound of formula I or a salt thereof exists in stereoisomeric forms (e.g., which contain one or more asymmetric carbon atoms), individual stereoisomers (enantiomers and diastereomers) and mixtures thereof are included within the scope of the invention. The invention also includes individual isomers of the compounds or salts represented by formula I, as well as mixtures of isomers with one or more chiral centers reversed therein. The scope of the invention includes: mixtures of stereoisomers, and purified enantiomerically or enantiomerically/diastereomerically enriched mixtures. The present invention includes mixtures of stereoisomers in all possible different combinations of all enantiomers and diastereomers. The present invention includes all combinations and subsets of stereoisomers of all specific groups defined above. The invention also includes geometric isomers, including cis-trans isomers, of the compounds of formula I or salts thereof.

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The reagents and starting materials used in the present invention are commercially available.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

The structures of all compounds of the invention can be determined by nuclear magnetic resonance¹H NMR) and/or mass spectrometric detection (MS).

¹H NMR chemical shifts (. delta.) are recorded by PPM (10)^-6). NMR was performed on a Bruker AVANCE-400 spectrometer. A suitable solvent is deuterated chloroform (CDCl)₃) Deuterated methanol (CD)₃OD), deuterated dimethyl sulfoxide (DMSO-d)₆) Tetramethylsilane was used as internal standard (TMS).

Low resolution Mass Spectrometry (MS) was determined by an Agilent 1200HPLC/6120 mass spectrometer using XBridge C18, 4.6 x 50mm, 3.5 μm, gradient elution conditions one: 80-5% of solvent A₁And 20-95% of solvent B₁(1.8 min)Bell), then 95% of solvent B₁And 5% of solvent A₁(over 3 minutes) as a volume percent of a solvent based on the total solvent volume. Solvent A₁: 0.01% trifluoroacetic acid (TFA) in water; solvent B₁: 0.01% trifluoroacetic acid in acetonitrile; the percentages are the volume percent of solute in solution. Gradient elution conditions two: 80-5% of solvent A₂And 20-95% of solvent B₂(1.5 min) and then 95% solvent B₂And 5% of solvent A₂(over 2 minutes) as a volume percent of a solvent based on the total solvent volume. Solvent A₂: 10mM ammonium bicarbonate in water; solvent B₂: and (3) acetonitrile.

All compounds of the invention can be separated by a high performance liquid chromatograph, a silica gel column chromatography and a thin layer chromatography flash column chromatography.

High performance liquid chromatography (prep-HPLC) liquid chromatography was prepared using shimadzu LC-20, column: waters xbridge Pre C18, 10um, 19mm 250 mm. The preparation conditions are as follows: mobile phase A: 0.05% aqueous trifluoroacetic acid (percentage is volume percent), mobile phase B: acetonitrile; gradient elution conditions: 25-75% of solvent A and 75-25% of solvent B; detection wavelength: 214nm, and/or 254nm, and/or 262 nm; flow rate: 10.0 mL/min.

Flash column chromatography (Flash system/Cheetah)^TM) Agela Technologies MP200 is used, and Flash column Silica-CS (80g) and Cat No. CS140080-0 are used as a matched separation column.

The thin layer chromatography is Nicotiana new chemical engineering, the thickness of the coating is 0.2 +/-0.03 mm, and the specification is 20 multiplied by 20 cm. The column chromatography is carried out by using 200-300 mesh silica gel of the yellow sea of Taiwan tobacco as carrier.

Example 1: synthesis of Compound 1.8

Step 1: to a solution of 2-amino-6-chloropyridine (40.0g, 311mmol) and lithium bistrimethylsilylamide (685mL, 685mmol, 1M in tetrahydrofuran) in tetrahydrofuran (400mL) was added di-tert-butyl dicarbonate (74.7g, 342mmol) dropwise under ice-bath conditions. The reaction system was stirred at room temperature overnight and then concentrated, diluted with ethyl acetate (400mL), the organic phase was washed with a hydrochloric acid solution (1M), a saturated aqueous sodium bicarbonate solution and a saturated brine, respectively, the organic phase was separated and dried over anhydrous sodium sulfate, filtered, concentrated, the resulting residue was recrystallized from ethanol, filtered, and the filter cake was dried to give compound 1.1(39.5g, yield: 56%) as a yellow solid.

Step 2: to a solution of compound 1.1(39.5g, 173mmol) and N, N, N ', N' -tetramethylethylenediamine (74.4g, 432mmol) in tetrahydrofuran (400mL) was added dropwise N-butyllithium (173mL, 432mmol, 2.5M N-hexane solution) under nitrogen at-78 ℃. After the addition was complete, the reaction was slowly raised to-10 ℃ and stirred at this temperature for 2 hours. The reaction was again cooled to-78 ℃ and N, N-dimethylformamide (25.3g, 347mmol) was added. After the addition was complete, the reaction was slowly warmed to room temperature and stirred overnight. The reaction was quenched with hydrochloric acid solution (1M) at-10 ℃ until pH 2-3, the aqueous phase was extracted with ethyl acetate (400mL), the organic phase was washed with water (200mL) and saturated brine (200mL), respectively, the separated organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 1.2(32.9g, yield: 74%) as a yellow solid.

And step 3: compound 1.2(32.9g, 128mmol) was added to a solution of hydrogen chloride in 1, 4-dioxane (300mL, 4M) and stirred at room temperature overnight. The solvent was removed by concentration under reduced pressure, the pH was adjusted to 7 to 8 with a saturated aqueous solution of sodium hydrogencarbonate, the aqueous phase was extracted with ethyl acetate (300mL × 2), the organic phases were combined and washed with water (200mL) and saturated brine (200mL), respectively, the separated organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound 1.3(18.9g, yield: 94%) as a yellow solid. M/z [ M + H ]]⁺157.0。

And 4, step 4: a mixed solution of compound 1.3(18.9g, 121mmol), compound 1.4A (46.8g, 121mmol) and toluene (400mL) was stirred under reflux overnight. The solvent was removed by concentration under reduced pressure, and the residue was recrystallized from ethanol (150mL), filtered, and the cake was dried under vacuum to give compound 1.4(13.1g, yield: 41%) as a yellow solid. M/z [ M + H ]]⁺266.0。

And 5: solution (80mL, 2M) and stirred at 30 ℃ for two days. The solvent was removed by concentration under reduced pressure, the pH was adjusted to 7 to 8 with a saturated aqueous sodium bicarbonate solution, the aqueous phase was extracted with a mixed solvent of dichloromethane and methanol (10:1, 200mL × 2), the organic phase was washed with water (100mL) and saturated brine (100mL), the separated organic phase was dried over anhydrous sodium sulfate, and the mixture was filtered and concentrated under reduced pressure to obtain compound 1.5(6.5g, yield: 98%) as a yellow solid. M/z [ M + H ]]⁺266.0，¹H NMR(400MHz,DMSO-d₆)δ7.90(d,J＝8.0Hz,1H),7.75(s,1H),7.39(br.s,2H),7.02(d,J＝8.0Hz,1H),4.24(q,J＝7.6Hz,2H),2.98(s,2H),1.29(t,J＝7.0Hz,3H)。

Step 6: to a solution of compound 1.5(6.5g, 24.5mmol) in dichloromethane (130mL) was added di-tert-butyl dicarbonate (13.4g, 61.2 mmol). The reaction system was stirred at room temperature for 5 days, and then diluted with dichloromethane (100mL), and the organic phase was washed with an aqueous citric acid solution (100mL), a saturated aqueous sodium bicarbonate solution (100mL) and a saturated brine (100 mL). The separated organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the obtained residue was purified by silica gel column chromatography (0% to 100% ethyl acetate petroleum ether solution) to obtain compound 1.6(8.6g, yield: 78%) as a yellow solid.

And 7: to a mixed solution of compound 1.6(4.0g, 10.9mmol) in tetrahydrofuran (21mL), ethanol (14mL) and water (7mL) was added lithium hydroxide monohydrate (1.38g, 32.8 mmol). The reaction system was stirred at room temperature overnight, poured into water, then adjusted to pH 4 with an aqueous citric acid solution, filtered, and the filter cake was washed with water and methyl t-butyl ether, respectively, and concentrated under reduced pressure to give compound 1.7(2.97g, yield: 80%) as an earth-yellow solid.

And 8: to N, N-dimethylformamide (30mL) was added sequentially the compound 1.7(2.97g, 8.8mmol), N, N-diisopropylethylamine (2.84g, 22mmol), di-N-propylamine (2.22g, 22mmol) and 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (7.13g, 22mmol) at-70 ℃. After the addition, the reaction system was stirred at 0 ℃ for 20 minutes, poured into water, extracted with ethyl acetate (100 mL. times.2), washed with saturated brine, and the organic phase was separated and washed with brineDried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (50% ethyl acetate in petroleum ether) to give compound 1.8(1.2g, yield: 32%) as a yellow oil. M/z [ M + H ]]⁺421.0。

Example 2: synthesis of Compound 2.11

Step 1: 3-bromo-2-chloro-6-methyl-5-nitropyridine (9.0g, 35.8mmol) and stannous chloride (27.2g, 143mmol) were added to ethanol (300mL), the reaction was warmed to 80 ℃ and stirred for 3 hours. The reaction mixture was concentrated under reduced pressure, diluted with ethyl acetate (400mL), the organic phase was washed with aqueous sodium hydroxide (1M), filtered, the filtrate was extracted with ethyl acetate (4X 100mL), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 2.1(7.3g, yield: 92%) as a yellow solid.

Step 2: compound 2.1(7.3g, 33.0mmol) was added to a mixed solution of acetic anhydride (40mL) and glacial acetic acid (20mL) and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to give compound 2.2(8.5g, yield: 98%) as an orange solid. M/z [ M + H ]]⁺263.0

And step 3: compound 2.2(8.5g, 32.3mmol) and magnesium sulfate (2.3g, 19.4mmol) were added to water (300mL), the system was heated to 80 deg.C, potassium permanganate (17.8g, 113mmol) was slowly added in portions, and the reaction was stirred at 80 deg.C overnight after the addition. Then, the reaction solution was cooled to room temperature, pH was adjusted to 1 with a 2% sulfuric acid aqueous solution, the reaction system was cooled to 10 ℃, a solid precipitated in the reaction solution, stirred for about 15 minutes, filtered, a filter cake was collected, and the filter cake was vacuum-dried to obtain compound 2.3(1.8g, yield: 19%) as a white solid. M/z [ M-H]^-291.0。

And 4, step 4: an aqueous solution of compound 2.3(1.6g, 5.8mmol) in sodium hydroxide (20mL, 1M) was stirred under reflux for 3.5 hours. Then, the reaction mixture was cooled to room temperature, acidified with concentrated hydrochloric acid, adjusted to pH 5, extracted with ethyl acetate (6 × 40mL), and the organic phases were combined and then extracted with ethyl acetateWashed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 2.4(1.2g, yield: 82%) as a white solid. M/z [ M + H ]]⁺251.0。

And 5: to a solution of compound 2.4(1.2g, 4.8mmol) in tetrahydrofuran (10mL) was added borane (28.6mL, 1M in tetrahydrofuran) dropwise at room temperature, and after addition, the reaction was allowed to stir at room temperature for an additional 3.5 hours. The reaction was quenched by addition of methanol, and the reaction was concentrated under reduced pressure to give compound 2.5(1.1g, yield 97%) as a yellow oily liquid. M/z [ M + Na]⁺258.9。

Step 6: compound 2.5(1.1g, 4.6mmol) and manganese dioxide (4.1g, 46.3mmol) were added to dichloromethane (40mL) and the reaction stirred vigorously at room temperature for 2.5 h. The reaction solution was filtered, the filter cake was washed with methylene chloride several times, the filtrates were combined and concentrated under reduced pressure to give compound 2.6(1.0g, yield: 92%) as a yellow solid.

And 7: compound 2.6(1.0g, 4.3mmol) and compound 1.4A (1.9g, 4.9mmol) were added to ethanol (20mL), the reaction was stirred under reflux for 5 hours, then the reaction liquid was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate 4/1 to 1/1) to give compound 2.7(750mg, yield: 51%) as a pale yellow solid. M/z [ M + H ]]⁺344.0。

And 8: compound 2.7(750mg, 2.2mmol) was added to a solution of hydrochloric acid in ethanol (10mL, 2M) and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure to give compound 2.8(600mg, yield: 80%) as a yellow solid. M/z [ M + H ]]⁺344.0，¹H NMR(400MHz,CD₃OD):δ8.17(d,J＝0.4Hz,1H),7.81(s,1H),4.35(d,J＝7.2Hz,2H),3.57(s,2H),1.38(t,J＝7.2Hz,3H)。

And step 9: compound 2.8(600mg, 1.7mmol), di-tert-butyl dicarbonate (2.0g, 8.7mmol) and triethylamine (1.1g, 10.5mmol) were added to dichloromethane (40mL) and stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether/ethyl acetate 4/1) to give compound 2.9(230mg, yield: 29%) as a bright yellow solid.

Step 10: compound 2.9(230mg, 0.5mmol) and oxyhydrogen monohydrateLithium (43.4mg, 1.0mmol) was added to a mixed solution of tetrahydrofuran (3mL) and water (3mL), and stirred at room temperature overnight. The reaction solution was acidified with 1N hydrochloric acid, pH was adjusted to 5, dichloromethane (8X 5mL) was added for extraction, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 2.10(200mg, yield: 92%) as a yellow solid. M/z [ M + H ]]⁺415.9。

Step 11: to a solution of compound 2.10(200mg, 0.5mmol) in dichloromethane (15mL) were added 1-hydroxybenzotriazole (97.3mg, 0.7mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (184mg, 0.9mmol), N, N-diisopropylethylamine (93mg, 0.7mmol) and di-N-propylamine (72.9mg, 0.7mmol) in that order, and after completion of the addition, the mixture was stirred at room temperature overnight. The reaction mixture was diluted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by prep-TLC (petroleum ether/ethyl acetate 4/1) to give compound 2.11(160mg, yield: 67%) as a yellow solid. M/z [ M + H ]]⁺499.0。

Example 3: synthesis of Compound 3.2

Step 1: a solution of 6-bromophthalazin-1 (2H) -one (0.80g, 3.55mmol), cesium carbonate (2.89g, 8.86mmol) and 2-chloromethyloxazole (0.50g, 4.25mmol) in N, N-dimethylformamide (10mL) was stirred at 50 ℃ for 5 hours, the reaction system was cooled to room temperature and then poured into water, the solid was collected by filtration, the filter cake was washed with water and petroleum ether, respectively, and dried under reduced pressure to give compound 3.1(0.84g, yield: 78%) as a yellow solid.

Step 2: under nitrogen protection, compound 3.1(500mg, 1.63mmol), potassium acetate (481mg, 4.9mmol), pinacol diboron (622mg, 2.45mmol) and Pd (dppf)₂Cl₂(11.9mg, 0.02mmol) of a mixture of 1, 4-dioxane (10mL) was stirred at 80 ℃ for 4 hours. Cooling the reaction system to room temperature, filtering with diatomite, leaching with ethyl acetate solution, concentrating the filtrate, pulping the obtained residue with ethanol,the solid was collected by filtration and dried under reduced pressure to give compound 3.2(528mg, yield: 92%) as a brown solid. M/z [ M + H ]]⁺354.0。

Example 4: synthesis of Compounds 3.3 to 3.8

Compounds 3.3-3.8 according to the embodiment 3 compound 3.2 synthesis method, using 6-bromine isoquinoline-1 (2H) -ketone, 6-bromine phthalazin-1 (2H) -ketone and corresponding chloride or bromide reaction to get:

example 5: synthesis of Compound 3.9

Compound 3.9 following the procedure for the synthesis of compound 3.2 of example 3, compound 3.2 in step 2 was replaced with 4- (pyrrolinylcarbonyl) bromobenzene to give:

example 6: synthesis of Compound 1-1-1

Step 1: to a solution of compound 1.8(120mg, 0.29mmol) in tetrahydrofuran (4mL) under nitrogen atmosphere were added compound 3.2(121mg, 0.34mmol), aqueous sodium carbonate (2mL, 2.0M) and [1,1' -bis (diphenylphosphino) ferrocene in that order]Palladium dichloride dichloromethane complex (23.3mg, 0.029mmol), after addition, the reaction was replaced three times with nitrogen and stirred at 70 ℃ for 25 minutes. The reaction was quenched by adding water (10mL) to the reaction system, the mixture was extracted with ethyl acetate (10 mL. times.2), the organic phases were combined, washed with saturated brine, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, and concentrated, and the obtained residue was purified by thin layer chromatography (10% methanol in dichloromethane) to give compound 1-1(50mg, yield: 29%) as a pale yellow solid. M/z [ M + H ]]⁺612.0。

Step 2: trifluoroacetic acid (1mL) was added dropwise to a solution of compound 1-1(50mg, 0.082mmol) in dichloromethane (2mL) in an ice-water bath, and after the addition was completed, the reaction was stirred at room temperature for 45 minutes. The solvent was removed under reduced pressure, and the obtained residue was purified by prep-HPLC to give compound 1-1-1(16.5mg, yield: 32%) as a white solid. M/z [ M + H ]]⁺511.5，¹H NMR(400MHz,CD₃OD):δ8.72-8.65(m,2H),8.50-8.44(m,2H),8.22-8.15(m,2H),7.93-7.89(m,1H),7.17-7.13(m,1H),7.11(s,1H),5.57(s,2H),3.48(br.s,4H),3.31(overlapping with solvent,2H),1.76-1.65(m,4H),0.95(br.s,6H)。

Example 7: synthesis of Compounds 1-1-2 to 1-1-4

Compounds 1-1-3-1-1-4 according to example 6 Synthesis of Compound 1-1-1, compound 3.2 in step 1 was replaced with 3.3-3.5 to give:

m/z:[M+H]⁺537.4，¹H NMR(400MHz,CD₃OD):δ8.69-8.62(m,2H),8.50-8.42(m,2H),8.16(s,2H),7.10(s,1H),4.73(t,J＝6.8Hz,2H),3.70(t,J＝6.8Hz,2H),3.47(br.s,4H),3.31(overlapping with solvent,2H),3.09(s,3H),1.76-1.64(m,4H),0.94(br.s,6H)。

m/z:[M+H]⁺523.4，¹H NMR(400MHz,CD₃OD):δ8.74-8.67(m,4H),8.51-8.45(m,2H),8.20(q,J＝8.2Hz,2H),7.39(t,J＝4.8Hz,1H),7.12(s,1H),5.67(s,2H),3.54-3.44(m,4H),3.31(overlapping with solvent,2H),2.09-1.97(m,4H),0.97(br.s,6H)。

m/z:[M+H]⁺550.8，¹H NMR(400MHz,CD₃OD):δ8.71(s,1H),8.70-8.66(m,1H),8.47(d,J＝9.4Hz,2H),8.23-8.16(m,2H),7.15(s,1H),7.11(s,1H),5.56(s,2H),3.49-3.39(m,4H),3.31(overlapping with solvent,2H),2.41(s,6H),1.75-1.66(m,4H),0.98(br.s,6H)。

example 8: synthesis of Compounds 1-2-1 to 1-2-3

Compound 1-2-1 to 1-2-3 according to the synthesis of Compound 1-1-1 of example 6, Compound 3.2 in step 1 was replaced with 3.6 to 3.8 to give:

m/z:[M+H]⁺510.8，¹H NMR(400MHz,CD₃OD):δ8.48-8.37(m,2H),8.35-8.28(m,1H),8.12(s,2H),7.94-7.88(m,1H),7.53(d,J＝7.2Hz,1H),7.15(s,1H),7.09(s,1H),6.85(d,J＝7.2Hz,1H),5.40(s,2H),3.49(br.s,4H),3.31(overlapping with solvent,2H),1.78-1.64(m,4H),0.96(br.s,6H)。

m/z:[M+H]⁺521.8，¹H NMR(400MHz,CD₃OD):δ8.72(d,J＝4.8Hz,2H),8.48(s,1H),8.40(d,J＝8.6Hz,1H),8.32(d,J＝8.6Hz,1H),8.15(s,2H),7.55(d,J＝7.4Hz,1H),7.38(t,J＝5.0Hz,1H),7.10(s,1H),6.86(d,J＝7.4Hz,1H),5.48(s,2H),3.48(s,4H),3.31(overlapping with solvent,2H),1.74-1.68(m,4H),0.96(br.s,6H)。

m/z:[M+H]⁺549.7，¹H NMR(400MHz,CD₃OD):δ8.47(s,1H),8.39(d,J＝8.8Hz,1H),8.33(d,J＝8.8Hz,1H),8.13(s,1H),7.53(d,J＝7.4Hz,1H),7.13(s,1H),7.09(s,1H),6.86(s,1H),6.84(s,1H),5.37(s,2H),3.50-3.45(m,4H),3.31(overlapping with solvent,2H),2.41(s,6H),1.75-1.65(m,4H),0.98(br.s,6H)。

example 9: synthesis of Compound 1-3-1

Compound 1-3-1 following the procedure for the synthesis of compound 1-1-1 of example 6, substituting compound 3.2 in step 1 with 3.9 gave:

m/z:[M+H]⁺459.9，¹H NMR(400MHz,CD₃OD):δ8.26(d,J＝8.4Hz,2H),8.09(d,J＝8.4Hz,1H),8.02(d,J＝8.4Hz,1H),7.68(d,J＝8.0Hz,2H),7.07(s,1H),3.63(t,J＝7.0Hz,2H),3.54-3.43(m,6H),3.31(overlapping with solvent,2H),2.06-1.98(m,2H),1.97-1.89(m,2H),1.75-1.64(m,4H),0.95(br.s,6H)。

example 10: synthesis of Compound 2-1-1

Step 1: compound 2.11(150mg, 0.3mmol), compound 3.5(120mg, 0.3mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium dichloromethane complex (15.0mg) were added to a mixed solution of 2M (mol/L) aqueous sodium bicarbonate solution (2mL) and tetrahydrofuran (6mL), the reaction system was replaced with nitrogen gas several times, and then heated to 70 ℃ and stirred for 1.2 hours. The reaction solution was cooled to room temperature, filtered through celite, and the filtrate was concentrated under reduced pressure and purified by prep-TLC (ethyl acetate) to give compound 2-1(130.0mg, yield: 69%) as an orange solid.

Step 2: trifluoroacetic acid (1.5mL) was slowly added dropwise to a solution of compound 2-1(30mg, 45.7mmol) in dichloromethane (1.5mL) at room temperature, and after completion of the addition, the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and recrystallized from ethanol to give compound 2-1-1(20mg, yield: 65%) as a white solid in the form of trifluoroacetate salt. M/z [ M + H ]]⁺556.9，¹H NMR(400MHz,CD₃OD):δ8.73(d,J＝5.0Hz,2H),8.48(t,J＝4.2Hz,2H),8.13(d,J＝1.4Hz,1H),8.03(dd,J＝8.4,1.7Hz,1H),7.91(s,1H),7.39(t,J＝5.0Hz,1H),7.03(s,1H),5.67(s,2H),3.63-3.39(m,6H),1.75-1.69(m,4H),0.97(br.s,6H)。

Example 11: synthesis of Compound 2-2-1

Compound 2-2-1 following the procedure for the synthesis of compound 2-1-1 in example 10, substituting compound 3.5 in step 1 with 3.3 gave:

m/z:[M+H]⁺570.9，¹H NMR(400MHz,CD₃OD):δ8.48(d,J＝10.4Hz,2H),8.10(s,1H),8.03(dd,J＝8.4,1.6Hz,1H),7.90(s,1H),7.02(s,1H),4.75(t,J＝6.8Hz,2H),3.71(t,J＝6.8Hz,2H),3.54-3.42(m,4H),3.31(overlapping with solvent,2H),3.09(s,3H),1.75-1.66(m,4H),0.96(s,6H)。

example 12: synthesis of Compound 3-1-1

Step 1: compound 2-1(24mg, 36.5. mu. mol), tetrakis (triphenylphosphine) palladium (8.0mg), formic acid (8.4mg, 183. mu. mol) and triethylamine (59.2mg, 584. mu. mol) were added to N, N-dimethylformamide (8mL), and the reaction was replaced with nitrogen several times, then heated to 100 ℃ and stirred for 3.5 hours. The reaction was cooled to room temperature, filtered through celite, and the filtrate was concentrated under reduced pressure to give compound 3-1(60mg, crude) as a yellow oily liquid. M/z [ M + H ]]⁺623.0。

Step 2: trifluoroacetic acid (1mL) was slowly added dropwise to a solution of compound 2-1(60mg, crude) in dichloromethane (1mL) at room temperature, and after the addition was complete, the reaction was stirred at room temperature for 1.5 hours. The reaction solution was concentrated under reduced pressure, and purified by prep-HPLC (acid method) to give compound 3-1(10mg, two-step yield: 52%) as a white solid. M/z [ M + H ]]⁺522.9，¹H NMR(400MHz,CD₃OD):δ8.74-8.69(m,3H),8.50(s,1H),8.47(d,J＝8.4Hz,1H),8.32(d,J＝1.6Hz,1H),8.24(dd,J＝8.4,1.8Hz,1H),7.95(d,J＝2.0Hz,1H),7.38(t,J＝5.0Hz,1H),6.96(s,1H),5.66(s,2H),3.49-3.43(m,4H),1.74-1.66(m,4H),0.94(d,J＝34.1Hz,6H)。

Example 13: synthesis of Compound 3-2-1

Compound 2-2 compound 3.5 in step 1 was replaced with 3.3 according to the synthesis of compound 2-1 of example 10. Compound 3-2-1 according to the synthesis of Compound 3-1-1 of example 12, Compound 2-1 in step 1 was replaced with 2-2. 3-2-1: m/z [ M + H ]]⁺536.9，¹H NMR(400MHz,CD₃OD):δ9.06(d,J＝2.0Hz,1H),8.48(t,J＝4.0Hz,2H),8.31(d,J＝1.2Hz,1H),8.24(dd,J＝8.4,1.6Hz,1H),8.19(d,J＝1.8Hz,1H),7.09(s,1H),4.73(t,J＝6.8Hz,2H),3.70(t,J＝6.8Hz,2H),3.60-3.38(m,5H),3.09(s,3H),1.77-1.66(m,4H),0.96(s,6H)。

Biological test examples

Example 1: cellular activity assay for TLR8

In this experiment, the determination of TLR8 bioactivity of a compound of formula I was tested using a cellular assay. The method is carried out in human embryonic kidney cells (HEK293) expressing TLR8 or other TLR family members such as TLR4, TLR7 and TLR9, after TLR agonist activates TLR, downstream NF-kB activation is caused, and then Secretory Embryonic Alkaline Phosphatase (SEAP) reporter gene is activated, and SEAP activity is detected by Quanti-Blue (InvivoGen) reagent, thereby reflecting the activity of TLR8 agonist.

The detailed experimental method is as follows:

HEK-BLUE-hTLR8 cell line was purchased from Invivogen, and cultured in DMEM medium containing 4.5g/L glucose (Sigma-Aldrich) and 10% fetal bovine serum at 37 deg.C, 95% humidity and 5% CO₂。

The test concentration of compound was from 0.5nM to 15. mu.M for 10 concentration gradients. TLR8 agonist of known activity was added as a positive control, and 1 μ L DMSO as a negative control.

The cells were treated as follows: the cells were removed from the culture dish and centrifuged to remove the medium, resuspended in a T-150 flask with 10mL of pre-warmed PBS, 12mL of pre-warmed medium was added, gently pipetted up and down, and counted under the microscope. 200,000 single cell suspensions per mL were immediately prepared in medium and 200. mu.L/well (40,000/well) were added to 96-well plates. The final concentration of DMSO was 0.5%.

The compound was added and incubated at 37 ℃ in a 5% CO2 incubator for 24 hours.

Pipette 20. mu.L/well of supernatant into 180. mu.L of 37 ℃ preheated Quanti-Blue, incubate at 37 ℃ for 1.5hr, and detect absorbance (OD value) with a spectrophotometer at 650 nm. The calculation formula of the activation effect is as follows:

percent effect ═ 100 (mean OD of dosing group-mean OD of DMSO group)/(mean OD of positive drug group-mean OD of DMSO group) × 100

The concentration-effect curve was fitted with Graphpad software and EC50 was calculated.

EC₅₀The range of value reports is: + represents 1-10 μ M, + represents 0.1-1 μ M, and +++ represents<0.1μM。

Compound numbering	TLR8EC50
		1-1-1	+++
1-1-2	++
		1-1-3	+++
1-1-4	+++
		1-2-1	++
1-2-2	+++
		1-2-3	+++
3-1-1	+++
		Positive control	++

Note: the positive control is VTX-2337(motolimod), chemical name: 2-amino-N, N-dipropyl-8- (4- (pyrrolidine-1-carbonyl) phenyl) -3H-benzo [ b ] azepine-4-carboxamide.

Claims (11)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof;

wherein X is N or CR₄(ii) a Y is N or CR₄(ii) a And X and Y are not CR at the same time₄；

L is a connecting bond;

r is a 9-15 membered bicyclic or tricyclic fused ring group, one ring is an aromatic ring, the other 1-2 rings are non-aromatic rings, the 9-15 membered bicyclic or tricyclic fused ring group includes 1-3N atoms in the ring atoms, and the non-aromatic ring further includes 1-2 oxo groups

And/or thio groups

R is unsubstituted or selectively substituted by 1-4R₅Substitution at any position: hydrogen, halogen, C_1-6Alkyl, halo C_1-6Alkyl, halo C_1-6Alkoxy, -L₁-Cy¹、-SR_d、-OR_d、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(＝NH)R_e、-C(＝NH)NR_dR_e、-S(O)_1-2R_eor-S (O)₂NR_dR_e(ii) a Wherein said C_1-6The alkyl is unsubstituted or optionally selected from-CN, -SR by 1-3_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(O)NR_dS(O)₂R_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-NR_dS(O)₂R_e、-NR_dS(O)₂NR_dR_e、-S(O)_1-2R_e、-S(O)₂NR_dR_e、-S(O)(＝NR_d)R_eand-P (O) (OR)_d)₂Substituted at any position;

L₁is a bond or- (CR)_aR_b)_n-；

Cy¹Is 3-10 membered heterocycloalkyl or 5-10 membered heteroaryl; the Cy is¹Is unsubstituted or optionally substituted by 1 to 3 groups selected from F, Cl, Br, C_1-4Alkyl, halo C_1-3Alkyl and halo C_1-3The radical of alkoxy is substituted in any position;

R₁and R₂Are each independently hydrogen or C_1-6An alkyl group;

or, R₁And R₂Together with the N atom to which they are both attached form a 3-8 membered heterocycloalkyl group; the heterocycloalkyl group may further contain 1 to 3 hetero atoms selected from N, O, S;

R₃and R₄Each independently is hydrogen, deuterium, halogen, amino, cyano, nitro, C_1-6Alkyl, halo C_1-6Alkyl, halo C_1-6Alkoxy radical, C_1-6Alkoxy or hydroxy C_1-6An alkyl group;

R_aand R_bEach independently is hydrogen, deuterium, halogen or C_1-6An alkyl group;

or, R_aAnd R_bTogether with the C atom to which they are jointly attached form C_3-8Cycloalkyl or 3-8 membered heterocycloalkyl;

each R_dAnd each R_eAre each independently hydrogen or C_1-6An alkyl group; the R is_dOr R_eIs unsubstituted or optionally substituted by 1 to 3 groups selected from halogen, hydroxy, amino, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Alkylamino, halogeno C_1-6Alkyl and halo C_1-6The substituent of the alkoxy is substituted at any position;

or, R_dAnd R_eTogether with the N atom to which they are both attached form a 3-8 membered heterocycloalkyl group; the heterocycloalkyl group may further contain 1 to 3 hetero atoms selected from N, O, S;

R_d1and R_e1Are each independently hydrogen or C_1-6An alkyl group;

n is an integer of 1 to 6.

2. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein R is₁And R₂Each independently is n-propyl;

and/or, R₃Is H, D, F, Cl, Br, -CN, -NH₂、-CH₃、-OCH₃、-OCF₃、-CH₂F、-CHF₂or-CF₃；

And/or, R₄Is H.

3. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 1, wherein R is any one of the following structures:

wherein R is₅Is hydrogen, halogen, C_1-6Alkyl, halo C_1-6Alkyl, halo C_1-6Alkoxy, -L₁-Cy¹、-SR_d、-OR_d、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(＝NH)R_e、-C(＝NH)NR_dR_e、-S(O)₂R_eand-S (O)₂NR_dR_e(ii) a Wherein said C_1-6The alkyl is unsubstituted or optionally substituted by 1 to 3 groups selected from-CN, -SR_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-C(O)NR_dS(O)₂R_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-NR_dS(O)₂R_e、-NR_dS(O)₂NR_dR_e、-S(O)_1-2R_e、-S(O)₂NR_dR_e、-S(O)(＝NR_d)R_eand-P (O) (OR)_d)₂Substituted at any position;

wherein L is₁、Cy¹、R_dAnd R_eIs as defined in claim 1.

4. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 3, wherein R is₅Is C_1-6Alkyl or-L₁-Cy¹(ii) a Said C is_1-6The alkyl is unsubstituted or optionally substituted by 1 to 3 groups selected from-SR_d、-OR_d、-OC(O)R_d、-OC(O)OR_d、-OC(O)NR_dR_e、-C(O)OR_d、-C(O)R_d、-C(O)NR_dR_e、-NR_dR_e、-NR_dC(O)R_e、-N(R_d)C(O)OR_e、-N(R_d)C(O)NR_dR_e、-NR_dS(O)₂R_e、-NR_dC(＝NH)R_e、-NR_dC(＝NH)NR_dR_e、-S(O)_1-2R_e、-S(O)₂NR_dR_e、-S(O)(＝NR_d)R_eand-NR_dS(O)₂NR_dR_eSubstituted at any position; r_dAnd R_eAre each independently hydrogen or C_1-6An alkyl group;

L₁is a connecting bond, -CH₂-、-CH₂CH₂-、-CH₂CH₂CH₂-、-CH₂CH₂C(CH₃)₂-、-CH₂CH₂CH₂CH₂-, or-CH₂CH(CH₃)CH₂-；

Cy¹Is a 5-6 membered heteroaryl; the Cy is¹Is unsubstituted or optionally substituted by 1 to 3 groups selected from F, Cl, Br, C_1-4Alkyl, halo C_1-3Alkyl and halo C_1-3The substituent of the alkoxy group is substituted at an arbitrary position.

5. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 3, wherein R is₅Is propyl, isopropyl, butyl, isobutyl, n-butyl, neopentyl, tert-amyl,

6. The compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 5, which is a compound of formula (II) or a pharmaceutically acceptable salt thereof:

wherein, U is CH or N; v is O or S;

x is N and Y is CH, or X is CH and Y is N;

R₅is as defined in any one of claims 1 to 5.

7. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, which is any one of the following compounds:

8. a pharmaceutical composition comprising a therapeutically effective amount of an active ingredient and a pharmaceutically acceptable adjuvant; the active component comprises the compound as shown in the formula (I) or pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7.

9. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, or a pharmaceutical composition as claimed in claim 8, in the preparation of a TLR8 modulator.

10. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 7, or a pharmaceutical composition as claimed in claim 8, in the manufacture of a medicament for the treatment, alleviation and/or prevention of a disease associated with TLR 8-mediated conditions.

11. The use as claimed in claim 10 wherein the TLR 8-mediated related disease is cancer, viral infection, inflammation, autoimmune disease, transplant rejection or graft versus host disease.