CN107296807A - Application of MALT1 targeting inhibitors in the preparation of MALT1-dependent tumor therapeutic drugs - Google Patents

Abstract

The invention relates to the application of MALT1 targeting inhibitors in the preparation of MALT1-dependent tumor treatment drugs. The present inventor screened out a compound with the function of inhibiting MALT1 from a large number of compounds, which provides a potential drug for the clinical treatment of MALT1-dependent tumors such as diffuse large B-cell lymphoma.

Description

MALT1-targeted inhibitors in the development of MALT1-dependent tumors Application in Therapeutic Drugs

technical field

The invention belongs to the field of biomedicine, and more specifically, the invention relates to the application of MALT1 targeting inhibitors in the preparation of MALT1-dependent tumor treatment drugs.

Background technique

Diffuse large B cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma, which can be divided into three subtypes according to its gene expression profile: germinal center B cell type ( Germinal center B cell like DLBCL (GCB-DLBCL), activated B cell like DLBCL (ABC-DLBCL) and primary mediastinal B cell lymphoma (PMBL). Among them, ABC-DLBCL has the highest degree of malignancy and is strongly resistant to existing immunochemotherapy. After classic R-CHOP treatment, the five-year survival rate of GCB-DLBCL can reach 76%, while the five-year survival rate of ABC-DLBCL is only about 30%. Therefore, it is important to find effective drugs and treatments for ABC-DLBCL Clinical problems that need to be solved urgently. Studies have shown that the important feature of ABC-DLBCL is the continuous activation of the NF-κB signaling pathway. Since the activation of this signaling pathway can promote the proliferation and survival of B cells and inhibit apoptosis in B cells, its continuous activation may be the cause of An important reason for the poor prognosis of ABC-DLBCL subtypes, suggesting that inhibiting the sustained activation of NF-κB signaling is the key to the treatment of ABC-DLBCL. However, since NF-κB is widely expressed in various tissues and organs, it is a transcription factor with extensive and important functions. After being activated, it enters the nucleus and binds to target DNA, participates in regulating the transcription and expression of a large number of genes, and directly inhibits NF-κB. It will lead to severe toxic side effects, so NF-κB itself is not suitable as a drug treatment-related target.

Studies have shown that the CBM complex composed of CARMA1 (also known as CARD11), BCL10, and MALT1 plays an important role in mediating NF-κB activation in lymphocytes. Mutations or translocations of CARMA1, BCL10, and MALT1 genes are often seen in ABC-DLBCL tumor cells, leading to enhanced or sustained activation of CARMA1, BCL10, and MALT1, which in turn causes excessive activation of NF-κB and tumorigenesis . RNA interference experiments show that the CBM complex is very important for the survival and proliferation of ABC-DLBCL cells, and inhibiting the expression of CBM can selectively kill ABC-DLBCL cells (V.N.Ngo et al., A loss-of-function RNAinterference screen for molecular targets in cancer., Nature, vol.441, no.7089, pp.106–110, 2006). Therefore, selectively blocking CBM signaling would be a very promising strategy for the treatment of ABC-DLBCL. In the CBM complex, both CARMA1 and BCL10 are scaffolding proteins without enzymatic activity, while MALT1 is a protease similar to caspase, which can inactivate negative regulators of NF-κB such as A20 and CYLD through its enzymatic cleavage activity, thereby Promote the activation of NF-κB (I.S.Afonina et al., MALT1-a universal soldier: multiple strategies to ensure NF-κB activation and target gene expression., FEBS J., vol.282, pp.1-12, 2015). In ABC-DLBCL, MALT1 is often continuously activated and leads to the continuous activation of NF-κB signaling pathway, specifically inhibiting the activity of MALT1 can reduce the production of growth factors and tumor suppressors, resulting in ABC-DLBCL cell death and Growth inhibition (U.Ferch et al., Inhibition of MALT1 protease activity is selectively toxic for activated Bcell-like diffuse large B cell lymphoma cells., J.Exp.Med., vol.206, no.11, pp.2313-2320, 2009; S. Hailfinger et al., Essential role of MALT1 protease activity inactivated B cell-like diffuse large B-cell lymphoma., Proc. Natl. Acad. Sci. U.S.A., vol.106, no.47, pp.19946-19951, 2009). More importantly, MALT1 knockout mice have no special phenotype and do not cause severe physiological damage to mice (A.A. Ruefli-Brasse et al. Regulation of NF-κB-Dependent Lymphocyte Activation and Development by Paracaspase,"Science(80- .)., vol.302, no.5650, pp.1581-1584, 2003), indicating that specifically inhibiting MALT1 will not cause serious side effects to patients. Therefore, MALT1 is a very potential treatment for ABC-DLBCL safe and effective drug targets.

In recent years, finding a targeted inhibitor of MALT1 for the treatment of ABC-DLBCL has become a research hotspot. Although some inhibitors have been discovered, their own shortcomings limit the further development and utilization of these inhibitors as clinical drugs. Z-VRPR-FMK is a polypeptide inhibitor that can specifically and irreversibly inhibit the activity of MALT1 (F.Rebeaud et al., The proteolytic activity of the paracaspase MALT1 is key in T cell activation., Nat.Immunol., vol.9 , no.3, pp.272-281, 2008), but its cell permeability is not high; phenothiazine derivatives are allosteric inhibitors of MALT1, but because they are also dopamine receptor antagonists, so May produce side effects such as off-target effects; MI-2 is an important small molecule inhibitor of MALT1 discovered in recent years, which can irreversibly inhibit the enzyme activity (L.Fontan et al., MALT1 smallmolecule inhibitors specifically suppress ABC-DLBCL in vitro and in vivo., Cancer Cell, vol.22, no.6, pp.812-24, 2012), but due to its solubility and stability, it still needs further optimization; and β-lapachone can inhibit The activity of MALT1, but its half-growth inhibitory concentration (IG50) to ABC-DLBCL cells is 1-10uM, and the killing or inhibition needs to be further optimized and improved (S.M.Lim et al., Identification of β-Lapachone Analogs as Novel MALT1 Inhibitors To Treat an AggressiveSubtype of Diffuse Large B-Cell Lymphoma, J.Med.Chem., p.acs.jmedchem.5b01415, 2015).

Therefore, finding novel MALT1 inhibitors is of great significance for the targeted therapy of malignant tumor ABC-DLBCL.

Contents of the invention

The purpose of the present invention is to provide the application of MALT1 targeting inhibitor in the preparation of MALT1 activity-dependent tumors such as diffuse large B-cell lymphoma therapeutic drugs.

In the first aspect of the present invention, the use of a compound or a pharmaceutically acceptable salt or precursor thereof in the preparation of a MALT1 inhibitor is provided; wherein the compound is:

In another preferred example, the MALT1 inhibitor prevents or treats MALT1 activity-dependent tumors.

In another preferred example, the MALT1 activity-dependent tumor includes but not limited to activated B-cell type (ABC-DLBCL) diffuse large B-cell lymphoma.

In another preferred example, the compound inhibits MALT1, thereby inhibiting CARMA1 or its family protein, the CBM complex composed of BCL10 and MALT1, so as to play a preventive or therapeutic role.

In another preferred example, the CARMA1 family proteins include (but not limited to): CARMA2, CARMA3, and CARD9.

In another aspect of the present invention, a method for preparing a MALT1 inhibitor is provided, the method comprising: mixing a compound with a pharmaceutically acceptable carrier to obtain a MALT1 inhibitor; the structural formula of the compound is:

Other aspects of the invention will be apparent to those skilled in the art from the disclosure herein.

Description of drawings

Figure 1. Gel filtration chromatography peak shape of LZ-MALT1 and SDS-PAGE results of peak products.

Figure 2. The inhibitory effect of positive control Z-VRPR-FMK on MALT1.

Fig. 3. The dose-response curve of small molecule compound WX28 inhibiting MALT1.

Figure 4. The growth inhibitory effect of WX28 on DLBCL cells.

detailed description

Through in-depth research and screening, the present inventor screened out a compound with the function of inhibiting MALT1 from a large number of compounds, and the compound has a significant inhibitory effect. The invention provides a potential drug for clinical treatment of MALT1-dependent tumors such as diffuse large B-cell lymphoma.

Compounds and their uses

It has been determined in the art that MALT1 is an enzyme similar to Caspase, and its protease cleavage activity is very important for the survival of tumor cells in malignant tumor diffuse large B-cell lymphoma, especially ABC-DLBCL lymphoma, and inhibiting its activity can be effective It can selectively inhibit and kill ABC-DLBCL tumor cells. Therefore, it was confirmed that a substance that inhibits MALT1 (MALT1 inhibitor) can prevent or treat diffuse large B-cell lymphoma, particularly ABC-DLBCL type lymphoma.

The inventors previously established a method for rapidly and effectively detecting MALT1 enzyme activity, and applied it to high-throughput screening of MALT1 small molecule inhibitors. Through high-throughput screening of 30,000 small molecule compounds, the inventors screened and further verified and confirmed a small molecule compound with an IC50 of less than 1 nM and a good dose-response curve. The small molecule compound is called WX28, and its structural formula is as follows:

The present invention also includes isomers, solvates, precursors of WX28, or their pharmaceutically acceptable salts, as long as they have the same or substantially the same function as WX28. The "pharmaceutically acceptable salt" refers to the salt formed by the reaction of the compound with inorganic acid, organic acid, alkali metal or alkaline earth metal, etc. These salts include (but are not limited to): (1) salts formed with inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid; (2) salts formed with organic acids such as acetic acid, oxalic acid, succinic acid, tartaric acid , methanesulfonic acid, maleic acid, or arginine. Other salts include those formed with alkali or alkaline earth metals such as sodium, potassium, calcium or magnesium, in the form of esters, carbamates, or other conventional "prodrugs". Compounds possess one or more asymmetric centers. These compounds may thus exist as racemic mixtures, individual enantiomers, individual diastereoisomers, diastereomeric mixtures, cis or trans isomers.

The "precursor of the compound" refers to that the precursor of the compound undergoes metabolism or chemical reaction in the patient's body to transform into the compound of the present invention, or a salt or solution containing the compound of the present invention when administered in an appropriate way.

Those skilled in the art should understand that after knowing the structure of the compound of the present invention, the compound of the present invention can be obtained by various methods well known in the art and using known raw materials, such as chemical synthesis or from organisms (such as animals or plants) ), these methods are included in the present invention.

The synthesized compound can be further purified by column chromatography, high performance liquid chromatography and the like.

Based on the inventor's new discovery, the present invention provides the use of the compound of the present invention or its isomer, solvate, precursor, or their pharmaceutically acceptable salts for the preparation of MALT1 inhibitors.

The MALT1 inhibitor prevents or treats MALT1 activity-dependent tumors such as diffuse large B-cell lymphoma, especially activated B-cell type (ABC-DLBCL) diffuse large B-cell lymphoma.

As used herein, the term "MALT1-dependent tumor" is used interchangeably with "MALT1 activity-dependent tumor" or "tumor dependent on MALT1 activity".

pharmaceutical composition

The present invention also provides a pharmaceutical composition for inhibiting MALT1, comprising: (a) an effective amount of the WX28 compound of the present invention, or its isomer, solvate, precursor, or their pharmaceutically an acceptable salt; and (b) a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition can be used to prevent or treat MALT1-dependent tumors such as diffuse large B-cell lymphoma, especially activated B-cell type (ABC-DLBCL) diffuse large B-cell lymphoma.

In the present invention, a "pharmaceutically acceptable" ingredient is a substance suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation and allergic reactions), ie having a reasonable benefit/risk ratio.

In the present invention, "pharmaceutically acceptable carrier" is a pharmaceutical carrier used to deliver the compound of the present invention, its isomer, solvate, precursor, or their pharmaceutically acceptable salt to animals or humans. Or food acceptable solvents, suspending agents or excipients. The carrier can be liquid or solid.

In the present invention, the pharmaceutical composition contains 0.0001-50% by weight of the compound of the present invention, its isomer, solvate, precursor, or their pharmaceutically acceptable salt. Preferably, the pharmaceutical composition contains 0.001-20% by weight of the compound of the present invention, its isomer, solvate, precursor, or their pharmaceutically acceptable salt. The effective administered dose of the compound of the present invention as an active ingredient may vary with the mode of administration and the severity of the disease to be treated.

The dosage form of the pharmaceutical composition of the present invention can be various, as long as it is a dosage form that can make the active ingredient effectively reach the body of a mammal. For example, it can be powder injection or injection. According to the convenience and needs of treatment, those skilled in the art can choose a dosage form that is convenient to use. The compounds of the present invention or pharmaceutical compositions thereof may also be stored in sterile devices suitable for injection or infusion.

screening method

The invention establishes a fluorescence luminescence method to detect the activity of MALT1, and performs high-throughput screening on more than 30,000 small molecule compounds. After screening and verification, the small molecular compound capable of effectively inhibiting the enzyme activity of MALT1 of the present invention is obtained.

Therefore, the present invention also provides a method for screening MALT1 inhibitors, said method comprising: (1) fusing the caspase-like domain fragment of MALT1 with the leucine zipper dimer fragment to obtain LZ-MALT1 recombinant fusion protein; (2) Contact the candidate substance with the recombinant fusion protein of step (1) and the MALT1 substrate; (3) Detect the fluorescence value of the AMC fluorescent group generated in the system of (2), if the fluorescence value occurs after adding the candidate substance Significantly lower, the candidate substance is a MALT1 inhibitor.

In a preferred mode of the present invention, when screening, in order to more easily observe the change of the interaction between the enzyme and the substrate, a control group can also be set, and the control group can be the said candidate substance without adding The interaction system of recombinant fusion protein and MALT1 substrate.

In a preferred mode of the present invention, during screening, a positive control is also set, and the fluorescence value generated by the positive control is used as a reference, thereby improving the accuracy of screening.

As a preferred mode of the present invention, the method also includes: further cell experiments and/or animal experiments are carried out on the obtained potential substances to further select and determine the effects on inhibiting MALT1 and further inhibiting MALT1-dependent tumors such as diffuse large B Substances with a more pronounced effect on cell lymphoma.

The potential substances for inhibiting MALT1 obtained by the screening method of the present invention can constitute a screening library, so that people can finally screen out substances that are more effective in inhibiting MALT1 and then inhibiting MALT1-dependent tumors such as diffuse large B-cell lymphoma. remarkable substance.

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Experimental methods not indicating specific conditions in the following examples are usually according to conventional conditions such as edited by J. Sambrook et al., Molecular Cloning Experiment Guide, Third Edition, Science Press, 2002, or according to the conditions described in the manufacturer suggested conditions.

Embodiment 1, LZ-MALT1 protein expression and purification

Because the full-length MALT1 and its caspase-like domain (340-789 amino acids) exist as monomers in physiological solution, the enzymatic cleavage activity is low, and MALT1 needs to form a dimer to better exert the enzymatic cleavage activity. After analysis and research, the inventors fused the caspase-like domain (340-789) fragment of MALT1 with the leucine zipper dimer fragment to construct an in vitro recombinant protein of LZ-MALT1.

The amino acid sequence of MALT1 (340-789) is as follows (SEQ ID NO: 1):

AKDKVALLIGNMNYREHPKLKAPLVDVYELTNLLRQLDFKVVSLLDLTEYEMRNAVDEFLLLLDKGVYGLLYYAGHGYENFGNSFMVPVDAPNPYRSENCLCVQNILKLMQEKETGLNVFLLDMCRKRNDYDDTIPILDALKVTANIVFGYATCQGAEAFEIQHSGLANGIFMKFLKDRLLEDKKITVLLDEVAEDMGKCHLTKGKQALEIRSSLSEKRALTDPIQGTEYSAESLVRNLQWAKAHELPESMCLKFDCGVQIQLGFAAEFSNVMIIYTSIVYKPPEIIMCDAYVTDFPLDLDIDPKDANKGTPEETGSYLVSKDLPKHCLYTRLSSLQKLKEHLVFTVCLSYQYSGLEDTVEDKQEVNVGKPLIAKLDMHRGLGRKTCFQTCLMSNGPYQSSAATSGGAGHYHSLQDPFHGVYHSHPGNPSNVTPADSCHCSRTPDAFISS

The leucine zipper dimer fragment is composed of amino acids of GCN4 (251-281), and the sequence is as follows, positions 3-33 ("HM" restriction enzyme site sequence in the following sequence):

HM KQLEDKVEELLSKNYHLENEVARLKKLVGER (SEQ ID NO: 2);

Between the GCN4 (251-281) zipper fragment and the MALT1 (340-789) fragment, there is a spacer fragment composed of "GSGSGSGS (SEQ ID NO: 3)", which is used to separate the two fragments to avoid steric hindrance . Formation of LZ-MALT1.

The amplified product of LZ-MALT1 (340-789) obtained above was inserted into the pET-28a plasmid (purchased from Novagen) at the Nde1/Not1 restriction site to obtain a recombinant plasmid containing the fusion fragment.

The recombinant plasmid obtained above was transformed into a Rosetta cell line for expression, and was initially purified by Ni-NTA (Qiagen, Valencia, CA) affinity chromatography, and then passed through a gel filtration chromatography column Superdex 200HR 10/300 (GE Healthcare, UK) After further purification, the target protein LZ-MALT1 was collected.

As shown in Figure 1, the peak of the target protein began to appear at about 12-14mL. SDS-PAGE gel was used to verify that the protein under this peak shape was indeed LZ-MALT1, with a purity of more than 95%. The obtained LZ-MALT1 protein was stored in a buffer containing 20 mM Tris (pH 7.5), 150 mM NaCl and 5 mM DTT.

Example 2, High-throughput screening of LZ-MALT1-specific small molecule inhibitors

The screening system established by the present inventors includes LZ-MALT1 and Ac-LVSR-AMC, wherein Ac-LVSR-AMC is the substrate of MALT1, MALT1 can recognize and cut the LVSR site, and the released AMC group can emit Fluorescence, so the enzymatic activity of MALT1 can be characterized by the detected fluorescence value. For determining the optimal high-throughput screening reaction conditions, the inventors set a series of LZ-MALT1 concentrations (25nM, 50nM, 100nM, 200nM, etc.) and substrate concentrations (62.5uM, 125uM, 250uM, etc.), and did two Two-dimensional orthogonal experiment, detect and record the fluorescence value every 30 seconds, the results are shown in Figure 2, the fluorescence value has a linear relationship with time, as the substrate concentration increases, the reaction signal increases, and as the enzyme concentration increases, the reaction signal It also increased accordingly, further proving that the purified LZ-MALT1 has good activity and can be used for subsequent high-throughput screening.

Ac-LVSR-AMC: Synthesized by Shanghai Qiangyao Biotechnology Co., Ltd.

The present inventor evaluates screening quality with Z factor, and Z factor is a kind of coefficient that evaluates data variation degree and response dynamic range, and calculation formula is: 1-3*(σ _p +σ _n )/(|μ _p -μ _n |), where σ _p/n is the standard deviation value of the positive/negative control, μ _p/n is the average value of the positive/negative control, and the Z factor greater than 0.5 means that the experiment can effectively distinguish signal from noise, which can be used for high-pass Quantitative screening (S.Inst and B.Parent, "Asimple statistical parameter for use in evaluation and validation of highthroughput screeining assays," J.Biomol.Screen., vol.4, no.2, pp.67–74, 1999) .

Considering the effect of high-throughput screening and the amount of enzyme and substrate, the inventors finally selected the combination of 25nMLZ-MALT1 and 80μM Ac-LVSR-AMCc, using Z-VRPR-FMK as a positive control, and using only the substrate BufferA was used as a negative control. After 90 minutes of reaction, the Z factor was 0.699, which was within the optimal range of 0.5-1.

Based on the above studies, the high-throughput screening reaction system was established as follows: Add the materials shown in Table 1 to Buffer A (20mM HEPES pH 7.5, 10mM KCl, 1.5mM MgCl ₂ , 1mM EDTA, 1mM DTT, 0.01% Triton X- 100), 20 μl/well.

Table 1

LZ-MALT1 25nM Ac-LVSR-AMC 80μM compound to be tested 10μM

The reaction was carried out in a 384-well plate (Greiner Bio One, Wemmel, Belgium, Catalog #784076), and the fluorescent signal of the reaction was detected with Envision Multilabel Reader (Perkin-Elmer, Waltham, MA). The emission wavelength and excitation wavelength were 360nm and 465nm, respectively. . In order not to miss slow-binding inhibitors, LZ-MALT1 and the test compound were pre-incubated at room temperature for 30 minutes, and then the substrate Ac-LVSR-AMC was added, at 0 minutes and 90 minutes after the addition of Ac-LVSR-AMC The signals are detected and recorded separately. In order to exclude the self-luminescence effect of the compound, the activity of MALT1 was characterized by the signal difference (T90-T0) at two time points.

The formula for the final inhibition rate is: [test compound fluorescence _(T90-T0) -negative control fluorescence _(T90-T0) ]/[positive control fluorescence _(T90-T0) -negative control fluorescence _(T90-T0) ]*100.

Among them, the known MALT1 inhibitor 50nM Z-VRPR-FMK was used as the positive control, and BufferA containing only the substrate was used as the negative control. Screening was performed with an inhibition rate of 40% as a threshold.

As a result, a total of 30007 compounds in the compound library (obtained from WuXi AppTec) were screened.

Embodiment 3, the effect verification of compound

For the small molecular compounds screened in Example 2, the effects of these compounds were verified by dose-response experiments, with the half inhibitory concentration (IC50) being less than 20 μM and having a good dose-response curve as the standard.

From these compounds, a small molecule compound with a strong inhibitory effect, an S-shaped dose-response curve, and an IC50 of less than 1 nM was obtained as a MALT1 inhibitor, as shown in Figure 3 and Table 2.

Table 2

The compounds in Table 2 can effectively inhibit the activity of MALT1 in vitro, are effective MALT1 inhibitors, and can be used as targeted drugs for the clinical treatment of ABC-DLBCL.

Example 4. Candidate compounds have specific growth inhibitory effects on ABC-DLBCL cell lines

The activity of MALT1 plays an important role in the proliferation of ABC-DLBCL cells, and the sensitivity of ABC-DLBCL cells and GCB-DLBCL cells to Z-VRPR-FMK, a positive inhibitor of MALT1, is different.

WX28 is the small molecule inhibitor with the lowest IC50 value and the best effect in the in vitro enzyme activity inhibition experiment. The inventors further verified its inhibitory effect through cell proliferation inhibition experiments.

In order to test whether the small molecule compound WX28 obtained by the screening of the present invention has a selective inhibitory effect on ABC-DLBCL, the inventors used different concentration gradients (0.01uM, 0.1uM, 1uM, 10uM, 100uM) of WX28 to act on two On ABC-DLBCL cell lines HBL-1, TMD8 and a GCB-DLBCL cell line OCI-LY1 (each cell line was purchased from ATCC), the number of cells was measured after 0 h and 72 h.

The results of the cell proliferation inhibition experiment showed that the compound WX28 can exert an inhibitory effect, and the ability to inhibit the ABC-DLBCL cell line TMD8 cells is the best, as shown in Figure 4.

Therefore, compound WX28 has the potential to inhibit cell growth efficiently.

discuss

ABC-DLBCL is currently the most malignant lymphoma, and it has strong drug resistance to the immunochemical R-CHOP therapy commonly used in clinical practice. One of its characteristics is the continuous activation of the NF-κB signaling pathway with growth-promoting and anti-apoptotic effects. Therefore, in theory, finding targeted drugs to block NF-κB signaling in ABC-DLBCL is an important strategy for the treatment of ABC-DLBCL. effective method. However, since NF-κB is a transcription factor with a wide range of important biological activities, it is not suitable as a therapeutic target by itself.

In B cells, antigen binding to the BCR induces Src family kinases to phosphorylate the tyrosines of the ITAM domains of CD79A and CD79B, and then the tyrosine kinase Syk is activated by binding to the phosphorylated ITAMs, thereby triggering the activation of the CD79A and CD79B ITAM domains. Signaling cascade reaction including Bruton's tyrosine kinase (BTK), phospholipase Cγ (phospholipase Cγ) and protein kinase Cβ (protein kinase Cβ, PKC-β). PKC-β phosphorylates CARM1, prompting it to recruit BCL10 and MALT1 to form a CBM complex, thereby activating IκB kinase (IKK), and finally stimulating the NF-κB signaling pathway. On the other hand, activated MALT1 cleaves and inactivates NF-κB negative feedback inhibitors such as A20 and CYLD through its enzymatic activity, further promoting the activation of NF-κB signaling pathway. Mutations or translocations of the above signaling pathway molecules are common in ABC-DLBCL tumor cells, such as CARMA1 mutations, CD79A/B mutations, BCL10 and MALT1 gene translocations, etc., leading to continuous activation of the NF-κB signaling pathway.

It can be seen from the above that the CBM complex plays an important role in mediating NF-κB activation. The only enzyme-active MALT1 protein in the CBM complex is a popular molecular target for the development of targeted drugs for the treatment of ABC-DLBCL in recent years: first, inhibiting the activity of MALT1 can effectively inhibit the signaling of NF-κB, thereby selectively inhibiting even Killing ABC-DLBCL cells, indicating the therapeutic effectiveness of targeting MALT1 enzyme activity; secondly, the MALT1 knockout mice showed some defects in the activation of T cells and B cells, but were phenotypically normal in other aspects, suggesting that the target The side effects of treating ABC-DLBCL with MALT1 enzyme activity will be relatively small; third, the caspase-like domain of MALT1 is unique in human genes, and inhibiting MALT1 will not produce extensive side effects caused by inhibiting other structurally similar proteins.

In recent years, the search for targeted inhibitors of MALT1 has become a research hotspot, but some existing compound inhibitors have different problems, poor druggability, and are difficult to be used for further clinical development. Therefore, the development of more effective MALT1 inhibitors with better druggability is of great significance for the treatment of ABC-DLBCL.

The inventors conducted high-throughput screening on 30,007 small molecules by establishing a high-throughput screening model, and obtained a compound WX28 with an IC50 of less than 1 nM. The invention provides a good foundation and guarantee for developing MALT1-targeted drugs to treat ABC-DLBCL.

All documents mentioned in this application are incorporated by reference in this application as if each were individually incorporated by reference. In addition, it should be understood that after reading the above teaching content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims (9)

1. the use of compound or its pharmaceutically acceptable salt or precursor in MALT1 inhibitor is prepared On the way；Wherein, the structural formula of described compound is：

2. purposes as claimed in claim 1, it is characterised in that described MALT1 inhibitor prevention Or treatment MALT1 dependent tumors.

3. purposes as claimed in claim 2, it is characterised in that described MALT1 dependent tumors Including：Diffusivity large B cell lymphoid tumor.

4. purposes as claimed in claim 1, it is characterised in that described diffusivity large B cell lymph Knurl is activating B cell type diffusivity large B cell lymphoid tumor.

5. the purposes as described in claim 1-4 is any, it is characterised in that described compound passes through suppression MALT1 processed, and then suppress the CBM of CARMA1 or its family protein, BCL10 and MALT1 compositions Compound, so as to play prevention or therapeutic action.

6. purposes as claimed in claim 5, it is characterised in that described CARMA1 family's egg It is white to include but is not limited to：CARMA2, CARMA3, CARD9.

7. a kind of method for preparing MALT1 inhibitor, it is characterised in that methods described includes：It will change Compound is mixed with pharmaceutically acceptable carrier, obtains MALT1 inhibitor；The knot of described compound Structure formula is：

8. method as claimed in claim 7, it is characterised in that described MALT1 inhibitor prevention Or treatment MALT1 dependent tumors.

9. method as claimed in claim 8, it is characterised in that described MALT1 dependent tumors Including but not limited to：Activating B cell type diffusivity large B cell lymphoid tumor.