CN115737645B

CN115737645B - Application of AT56 combined with sorafenib in peripheral T cell lymphoma

Info

Publication number: CN115737645B
Application number: CN202211355818.6A
Authority: CN
Inventors: 王欣; 胡顺凤; 周香香
Original assignee: Shandong Provincial Hospital Affiliated to Shandong First Medical University
Current assignee: Shandong Provincial Hospital Affiliated to Shandong First Medical University
Priority date: 2022-11-01
Filing date: 2022-11-01
Publication date: 2023-05-23
Anticipated expiration: 2042-11-01
Also published as: CN115737645A

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an application of AT56 combined with sorafenib in peripheral T cell lymphoma. The invention reports for the first time that the PTGDS specific inhibitor AT56 can significantly inhibit the growth of PTCL and has time dependence and concentration dependence. In vivo and in vitro experimental results demonstrate that AT56 promotes the iron death process by inducing iron ion aggregation in PTCL cells. Furthermore, AT56 in combination with sorafenib showed better anti-tumor effect in both PTCL cells and mouse models. The invention can promote the deep research and clinical application of the AT56 and sorafenib combined medication scheme, provides solid theoretical basis and experimental basis for constructing a novel efficient combined medication strategy and optimizing a PTCL clinical diagnosis and treatment scheme, and has good practical popularization and application values.

Description

Application of AT56 combined with sorafenib in peripheral T cell lymphoma

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an application of AT56 combined with sorafenib in peripheral T cell lymphoma.

Background

The disclosure of this background section is only intended to increase the understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.

Peripheral T Cell Lymphoma (PTCL) is a malignant tumor derived from abnormal proliferation of T cells at different stages after thymus, and belongs to a special subtype among non-Hodgkin lymphomas. PTCL has low incidence and regional differences. The disease has high invasiveness, high heterogeneity in biological behaviors, clinical manifestations, prognosis and the like, rapid disease progression and most of diagnosis in late stages. At present, the treatment of PTCL mainly takes radiotherapy and chemotherapy, the progress of first-line treatment in clinic is stopped for a long time, a specific and efficient targeted treatment means is lacked, the 5-year survival rate of patients is less than 30%, and a novel medicine and combined drug scheme is needed to be discovered clinically so as to improve the clinical benefit of PTCL patients.

Sorafenib (Sorafenib), a multi-target kinase inhibitor, was approved by the FDA in 2005 for the treatment of non-surgical or distant metastatic hepatocellular carcinoma, advanced renal carcinoma, and advanced refractory differentiated thyroid carcinoma with local recurrence or metastasis. In addition, sorafenib can induce iron death in tumor cells by inhibiting cystine/glutamate antiport protein xCT, thereby participating in the development and progression of tumors. Iron death is a novel mode of apoptosis that relies on intracellular iron accumulation and the toxic lipid peroxide ROS. More and more preclinical studies indicate that induction of iron death is an effective treatment strategy for tumor treatment and resistance to drug resistance. The current research finds that sorafenib plays a remarkable anti-tumor role in various non-hodgkin lymphomas, and one phase II clinical test result shows that sorafenib has a certain treatment effect and good tolerance in recurrent refractory lymphomas, but the anti-tumor activity of single drug treatment is lower. Thus, there is a need to explore new combination regimens based on sorafenib in depth to improve the clinical efficacy of lymphoma patients.

PTGDS is a protein with dual functions of catalyzing prostaglandin metabolism and lipid transport, and AT56 is a small molecule inhibitor that targets PTGDS proteins with high specificity. Currently, studies on PTGDS and AT56 are mainly focused on neurological diseases, inflammation, immune disorders, parasitic infections, and the like. Recent studies have found that PTGDS can play an important role in the development and progression of a variety of tumors by regulating biological functions such as cell proliferation, cell cycle, distant metastasis and stem cell characteristics. However, AT present, research on PTGDS and AT56 in PTCL has not been reported AT home and abroad.

Disclosure of Invention

Aiming AT the defects existing in the prior art, the invention provides the application of AT56 combined with sorafenib in peripheral T cell lymphoma. According to the invention, the PTGDS specific inhibitor AT56 can obviously inhibit the growth of PTCL, and AT56 combined with sorafenib shows a good anti-tumor effect in PTCL cells and a mouse model for the first time. The invention clarifies the combined anti-tumor effect of the PTGDS specific inhibitor AT56 and sorafenib, discusses the regulating action and molecular mechanism of AT56 on the iron death process in PTCL, and lays a solid theoretical foundation for deep exploration of novel PTCL medicines and combined medicine schemes. Based on the above results, the present invention has been completed.

Specifically, the invention relates to the following technical scheme:

in a first aspect of the invention, there is provided the use of AT56 in combination with sorafenib for the manufacture of a medicament for the treatment of peripheral T cell lymphoma.

According to the invention, the concept of "treatment" means any measure suitable for the treatment of tumors and related diseases, or for the prophylactic treatment of such a represented disease or of a represented symptom, or for the avoidance of recurrence of such a disease, for example after the end of a treatment period or for the treatment of a symptom of a disease that has already developed, or for the pre-interventional prevention or inhibition or reduction of the occurrence of such a disease or symptom.

As described above, the present invention has found through studies that AT56 can enhance the proliferation inhibitory ability of sorafenib on PTCL cells and enhance the promotion of lipid peroxidation, indicating that it has a promoting effect on iron death process in PTCL. Further animal experiment results show that AT56 combined with sorafenib shows good anti-tumor effect in a mouse model, and is obviously superior to single administration.

Accordingly, in a second aspect of the invention there is provided the use of an AT56 as described above in combination with sorafenib in any one or more of the following:

a) Inhibit proliferation of peripheral T cell lymphoma cells or preparing a product that inhibits proliferation of peripheral T cell lymphoma cells;

b) Promoting the death of peripheral T cell lymphoma cell iron or preparing a product for promoting the death of peripheral T cell lymphoma cell iron;

c) Inhibit peripheral T cell lymphoma growth or prepare a product for inhibiting peripheral T cell lymphoma growth.

Wherein in said b), promotion of iron death of peripheral T cell lymphoma cells is at least manifested as promotion of iron ion aggregation in tumor tissue (cells) and/or promotion of lipid peroxidation of tumor tissue (cells).

In a third aspect of the invention, a pharmaceutical composition is provided, the active ingredients of which comprise AT56 and sorafenib.

The pharmaceutical composition has the effect of treating peripheral T cell lymphoma, and is specifically expressed as follows:

a) Inhibit proliferation of peripheral T cell lymphoma cells;

b) Promote iron death of peripheral T cell lymphoma cells;

c) Inhibit peripheral T cell lymphoma growth.

In a fourth aspect of the invention, there is provided a method of treating peripheral T cell lymphoma, said method comprising: the above pharmaceutical composition is administered to a subject.

The beneficial technical effects of one or more of the technical schemes are as follows:

the technical scheme reports that the PTGDS specific inhibitor AT56 can significantly inhibit the growth of PTCL for the first time and has time dependence and concentration dependence. In vivo and in vitro experimental results demonstrate that AT56 promotes the iron death process by inducing iron ion aggregation in PTCL cells. Furthermore, AT56 in combination with sorafenib showed better anti-tumor effect in both PTCL cells and mouse models. The invention can promote the deep research and clinical application of the AT56 and sorafenib combined medication scheme, provides solid theoretical basis and experimental basis for constructing a novel efficient combined medication strategy and optimizing a PTCL clinical diagnosis and treatment scheme, and has good practical popularization and application values.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention.

FIG. 1 is a schematic diagram of molecular structures and molecular formulas of AT56 and sorafenib; wherein A is AT56 and B is sorafenib.

Fig. 2 shows that AT56 significantly inhibited the proliferation level of PTCL cells and that inhibition was concentration-dependent and time-dependent in the examples of the present invention.

FIG. 3 is a thermal graph showing the differential gene expression associated with iron death in AT56 treated and control treated PTCL cells in the quantitative determination of TMT protein in accordance with an embodiment of the present invention.

FIG. 4 shows the anti-lymphoma effect of AT56 (100. Mu.M) on an iron death agonist (5. Mu.M sorafenib and 5. Mu.M Erastin) in the examples of the present invention; wherein A is a proliferation inhibition result graph of PTCL by different treatment modes; b is a graph of lipid peroxidation promotion results of PTCL by different treatment modes.

FIG. 5 is a graph showing the combined anti-lymphoma effect of AT56 and sorafenib in a PTCL mouse xenograft model according to an embodiment of the present invention; wherein A is the tumor growth curve of mice in different treatment groups; b is a tumor picture after anatomical separation in mice of different treatment groups.

FIG. 6 shows the anti-lymphoma effect of iron death inhibitor Ferrostatin-1 in the examples of the present invention in part in response to AT 56; wherein A is the proliferation inhibition effect of the AT56 can be partially recovered after the Ferrositin-1 is added; b is that the addition of Ferrositin-1 can partially restore the promotion effect of AT56 on lipid peroxidation.

Fig. 7 shows the expression levels of iron death-related proteins in PTCL cells after AT56 treatment in examples of the present invention.

FIG. 8 shows the anti-lymphoma effect of the iron chelator DFO in part in response to AT56 in an embodiment of the invention; wherein, A is the proliferation inhibition of the partial recovery AT56 after adding DFO; b is the promotion of lipid peroxidation by the addition of DFO, which may be partially recovered by AT 56.

FIG. 9 is a graph showing the concentration levels of iron ions in tumor tissues of mice of different treatment groups in the examples of the present invention.

Detailed Description

It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present application. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments in accordance with the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.

In one exemplary embodiment of the invention, there is provided the use of AT56 in combination with sorafenib for the manufacture of a medicament for the treatment of peripheral T cell lymphoma.

Wherein the AT56 (C ₂₅ H ₂₇ N ₅ ) Is a potent, selective and orally active inhibitor of lipocalin prostaglandin D synthase with CAS number 162640-98-4, having the structural formula shown in FIG. 1A.

The sorafenib (C) ₂₁ H ₁₆ ClF ₃ N ₄ O ₃ ) Is a multi-target kinase inhibitor, with CAS number 284461-73-0, and its structural formula shown in FIG. 1B.

The molar ratio of AT56 to sorafenib is 1-20:1.

accordingly, in yet another embodiment of the present invention, there is provided the use of the above AT56 in combination with sorafenib in any one or more of the following:

The molar ratio of AT56 to sorafenib is 1-20:1.

a) In-c), the product may be a pharmaceutical or a test article, in particular a test agent, for use in basic research for constructing relevant tumor cells or animal models for studying e.g. iron death related mechanism problems in tumor cells.

When the product is a medicament, the medicament further comprises at least one non-pharmaceutically active ingredient. The non-pharmaceutically active ingredient comprises a pharmaceutically acceptable carrier, excipient and/or diluent.

In yet another embodiment of the present invention, a pharmaceutical composition is provided, the active ingredients of which comprise AT56 and sorafenib.

The molar ratio of AT56 to sorafenib is 1-20:1.

a) Inhibit proliferation of peripheral T cell lymphoma cells;

b) Promote iron death of peripheral T cell lymphoma cells;

c) Inhibit peripheral T cell lymphoma growth.

In yet another embodiment of the present invention, the pharmaceutical composition further comprises at least one non-pharmaceutically active ingredient. The non-pharmaceutically active ingredient comprises a pharmaceutically acceptable carrier, excipient and/or diluent.

In yet another embodiment of the present invention, the non-pharmaceutically active ingredient comprises:

pharmaceutically compatible inorganic or organic acids or bases, polymers, copolymers, block copolymers, monosaccharides, polysaccharides, ionic and nonionic surfactants or lipids; pharmacologically harmless salts (preferably sodium chloride), flavoring agents, vitamins (preferably vitamin a or vitamin E, tocopherol or provitamin), antioxidants (preferably ascorbic acid), and stabilizers and/or preservatives.

The administration forms of the pharmaceutical preparation include: liquid dosage forms, solid dosage forms, external preparations and sprays;

in yet another embodiment of the invention, the following dosage forms are included: true solutions, colloids, microparticle dosage forms, emulsion dosage forms, mixed rotation dosage forms, tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, freeze-dried powder injection, inclusion compounds, landfill agents, patches and liniment.

In yet another embodiment of the present invention, there is provided a method of treating peripheral T cell lymphoma comprising: the above pharmaceutical composition is administered to a subject.

The subject is an animal, preferably a mammal, most preferably a human, who has been the subject of treatment, observation or experiment.

The invention is further illustrated by the following examples, which are not to be construed as limiting the invention. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention.

Examples

Experimental method

1. Cell culture

The humanized PTCL cell lines Karpas-299 and Myla3676 used in the present study were purchased from ATCC, cultured in a 37℃cell incubator containing 5% CO2 using IMDM complete medium containing 10% fetal bovine serum, and the medium was replaced every 2-3 days and passaged, and cells with a good growth state were taken for subsequent experiments.

2.Cell counting kit-8 (CCK-8) detection of proliferation levels of PTCL cells

AT56, sorafenib, erastin, ferrostatin-1 (Fer-1) and Deferoxamine (DFO) powders were dissolved in DMSO to prepare respective stock solutions having concentrations of 125mM, 50mM, 10mM and 500 mM. PTCL cells in the logarithmic growth phase and in good condition were collected, the supernatant was removed after centrifugation, the cells were resuspended in IMDM complete medium and counted under a microscope per unitHole 10 ⁴ Inoculating PTCL cells to 96-well cell culture plate at a concentration of 90 μl, adding 10 μl of different concentrations of drugs according to concentration conversion formula, arranging blank hole and three repeated holes, mixing by gentle shaking, and placing 96-well plate in a solution containing 5% CO ₂ Continuously culturing in a 37 ℃ cell incubator; after 48-72 hours, 10 mu LCCK8 reagent is added into each hole, air bubbles are not required to be generated, after the mixture is incubated for 1-4 hours at 37 ℃ in a dark place, an enzyme-labeled instrument is used for detecting the absorbance value (OD value) of each hole at 450nm, and a growth curve of PTCL cells is drawn according to the detection result.

TMT (Tandem Mass Tag) quantitative protein detection and bioinformatics analysis

PTCL cells in the logarithmic growth phase and in good condition were collected at 3X 10 per well ⁵ PTCL cells were seeded onto 6-well cell culture plates AT 2mL, AT56 was added AT a final concentration of 100. Mu.M according to the concentration conversion formula, and a blank well and three duplicate wells were set. The 6-well plate was placed in a chamber containing 5% CO ₂ The culture was continued in a 37℃cell incubator for 48 hours, and the cells were collected. Protein extraction, quantification, enzyme digestion, isotope labeling, peptide enrichment and mass spectrometry detection are completed by Beijing nozaku source science and technology company. Biological information analysis is carried out according to TMT detection results, and the biological information analysis is carried out according to p<0.05、FC(Fold Change)>Screening for iron death-related differentially expressed proteins between the AT56 dosing group and the control group and mapping the heat.

Detection of lipid peroxidation in PTCL cells by C11-BODIPY method

PTCL cells grown well were resuspended using IMDM complete medium and seeded in 6-well plates at 3X 10 ⁵ Every 2mL of cells, adding the drug with the specified concentration, slightly shaking, and placing a 6-well plate at 37 ℃ with 5% CO ₂ Cells were collected after 48 hours incubation in an incubator. Diluting the C11-BODIPY probe with serum-free IMDM medium to a final concentration of 2.5. Mu.M, adding to PTCL cells, gently stirring to resuspend the cells, and standing at 37deg.C with 5% CO ₂ Incubate in incubator for 30 min in dark, gently invert mix every 5 min to allow sufficient contact of cells with probes. After the probe loading was completed, the cells were washed with serum-free IMDM medium, and repeated 3 times to remove the C11-BODIPY probe that did not enter the cellsA needle. Cells were resuspended using pre-chilled PBS, the cell suspension was filtered through a 400 mesh screen, and the cells were transferred to a labeled flow tube and the positive cell fraction was detected on-machine.

5. Construction of PTCL xenograft mouse model

Female 3-4 week old SCID Beign mice used in this study were purchased from Experimental animals, inc., vitreoscilla, beijing, and were kept in a sterile, constant temperature (25 ℃) and constant humidity (70%) environment using sterile drinking water, feed and litter. The animal research process strictly follows ARRIE rules and the rules of ethical committee for animal protection and research use in Shandong provincial hospitals. The experimental mice were divided into 4 groups according to a simple random method, a control group, an AT56 group, a sorafenib group, and an AT 56-combined sorafenib group, each group of 6 mice. PTCL cells in the logarithmic growth phase and in good condition were collected and the cell density was adjusted to 1X 10 ⁸ And adding an equal volume of Matrigel matrix gel per mL, uniformly mixing, taking 200 mu L of mixed solution, inoculating the mixed solution to the outer skin of the right lower limb of the mouse, forming an oval skin by naked eyes, and continuing to feed in a sterile environment. After the tumor is above the skin surface, the drug treatment is started, sorafenib (80 mg/kg) is administered by intraperitoneal injection, and AT56 (80 mg/kg) administration is realized by gastric lavage. Each day of vernier calipers measures tumor body size and records, a=tumor long diameter, b=tumor short diameter, tumor volume (V) = (a×b) ² ) X 0.5, and tumor growth curves are drawn. Mice were sacrificed 15 days later by cervical spining, and the complete subcutaneous tumors were dissected and stored in photographs. After the tissue of the mice is collected, the mice are respectively placed in 4% paraformaldehyde and an ultralow temperature refrigerator at the temperature of minus 80 ℃ for storage for later experiments.

6. Protein extraction and Western blotting analysis (Western blotting)

Collecting the PTCL cells after treatment, re-suspending and washing the cells by using precooled PBS, repeating for 3 times, centrifuging to remove the supernatant, adding a proper amount (40-150 mu L) of protein lysate (RIPA: PMSF: phosphatase inhibitor volume ratio is 100:1:1) according to the size of the cell mass), uniformly mixing by vortex oscillation, incubating on ice for 30 minutes, centrifuging at 12000rpm for 30 minutes at 4 ℃, sucking the supernatant containing protein, detecting the protein concentration by using a BCA method, and mixing the supernatant with a Loading Buffer according to volume 3:1, and carrying out metal bath at 100 ℃ for 10min to denature protein, and placing the mixture in a refrigerator at-80 ℃ for standby. According to the molecular weight of target protein, preparing 7.5-12% electrophoresis gel, placing in electrophoresis tank, adding proper quantity of electrophoresis buffer solution, adding 40 mug protein or 4 mug protein marker into each sample-adding hole in turn, making 200V constant pressure electrophoresis, and according to the positions of protein marker and bromophenol blue, determining electrophoresis time. PVDF film with the size corresponding to the gel is cut in advance and soaked in methanol for 1-2 minutes. Immersing the cut gel, PVDF membrane and transfer printing filter paper into a transfer film buffer solution, and respectively placing from bottom to top: transfer filter paper-PVDF film-gel-transfer filter paper, carefully remove bubbles, then put into a semi-dry transfer tank, transfer film for 30 minutes under the condition of constant pressure of 10V. PVDF membranes were placed in 10% skim milk formulated with TBST and blocked for 1-2 hours at room temperature. The PVDF membrane was then placed in a suitable concentration of primary antibody solution formulated with primary antibody diluent and incubated overnight at 4℃on a shaking table. After washing the PVDF membrane for 3 times by TBST, placing the PVDF membrane in a secondary antibody solution prepared by 10% skimmed milk, incubating the PVDF membrane for 1 hour at room temperature, washing the PVDF membrane again, dripping a developing solution (A solution: B solution=1:1) prepared in advance, imaging the PVDF membrane by using an Amersham Image 600 gel imaging instrument, analyzing the gray value of a strip by using Image J software, and evaluating the expression level of a target molecule.

7. Colorimetric detection of iron ion levels in mouse tissues

The iron ion detection kit was purchased from abcam company (cat# ab 8366). Fresh mouse tumor tissue was collected, washed 3 times with pre-chilled PBS, dried and weighed on a precision electronic balance. 100-200 mu L of buffer solution is added according to the weight of the tissue, the tissue is broken by ultrasonic vibration (5 minutes), the tissue is centrifuged for 10 minutes by 16000g of a centrifuge at 4 ℃, and the supernatant is collected and then placed on ice for storage. According to the instruction of the kit, 100 mu L of standard substance/tissue supernatant is added into a 96-well plate, balanced for 30 minutes in a constant-temperature incubator at 37 ℃,100 mu L of iron ion probe is added, mixed evenly by light shaking, and the mixture is further placed in the constant-temperature incubator at 37 ℃ to be incubated for 60 minutes in a dark place. The absorbance value (OD value) of each hole at 593nm is detected by using an enzyme-labeled instrument, and the concentration level of the iron ions in the tumor tissue of the mouse is calculated by drawing an iron ion concentration standard curve according to the absorbance value.

Experimental results:

1. FIG. 1 shows a schematic molecular structure and molecular formula of AT56 and sorafenib.

2. Figure 2 shows that AT56 significantly inhibited the proliferation level of PTCL cells and that inhibition was concentration-dependent and time-dependent. The results demonstrate the in vitro anti-lymphoma effect of AT 56.

3. FIG. 3 shows heat maps of differential gene expression associated with iron death in AT56 treated and control treated PTCL cells in TMT protein quantification. The results demonstrate the modulation of iron death-related gene expression levels in PTCL cells by AT 56.

4. Figure 4 shows that AT56 enhances the anti-lymphoma effect of iron death agonists (sorafenib and Erastin). In FIG. 4A, AT56 (100. Mu.M), sorafenib (5. Mu.M) and Erastin (5. Mu.M) alone inhibited the proliferation level of PTCL cells, and the proliferation inhibition capacity of sorafenib (5. Mu.M) and Erastin (5. Mu.M) was enhanced by the addition of AT56 (100. Mu.M). In FIG. 4B, the addition of AT56 (100. Mu.M) enhances the promotion of lipid peroxidation by sorafenib (5. Mu.M) and Erastin (5. Mu.M). The results demonstrate the promoting effect of AT56 on iron death processes in PTCL.

5. Fig. 5 shows the combined anti-lymphoma effect of AT56 and sorafenib in PTCL mouse xenograft model. Fig. 5A shows tumor growth curves for different groups of mice, where both AT56 and sorafenib alone inhibited lymphoma growth, and addition of AT56 enhanced sorafenib in vivo antitumor effects. Figure 5B shows images of tumors from different groups of mice after dissection, with the tumor size and volume of the combination group significantly smaller than the control group and the single drug treatment group. The results demonstrate the in vivo combined antitumor effect of AT56 and sorafenib.

6. FIG. 6 shows that the iron death inhibitor Ferrostatin-1 partially reverts to the anti-lymphoma effect of AT 56. In FIG. 6A, AT56 inhibits proliferation of PTCL cells, and the proliferation inhibiting effect of AT56 is partially recovered after the addition of Ferrostatin-1. In FIG. 6B, the addition of Ferrosistatin-1 partially reverted the promotion of lipid peroxidation by AT 56. The results indicate the dependence of the anti-tumor effect of AT56 on iron death in PTCL.

7. Fig. 7 shows the expression levels of iron death-related proteins in PTCL cells after AT56 treatment. As AT56 concentration increased, the expression level of iron-death-related transcription factor NRF2 decreased, and the expression level of negative regulator KEAP1 of NRF2 increased significantly, indicating activation of iron death process in PTCL cells. The iron death process is driven primarily by iron ion aggregation and lipid peroxidation, and we have further explored the driving factors for iron death in PTCL cells receiving AT56 treatment by conducting experiments. Western blotting showed that NCOA4, FTH and FTL expression levels were significantly elevated in PTCL cells receiving AT56 treatment, indicating abnormal aggregation of iron ions in PTCL cells. However, the expression level of the key enzyme for lipid metabolism, ACSL4, was decreased, and the expression levels of the oxidative regulatory molecules xCT and GPX4 were increased, indicating compensatory inhibition of lipid metabolism and peroxidation in PTCL cells. The results demonstrate potential modulatory effects of AT56 on iron death and iron ion aggregation.

8. Figure 8 shows that the iron chelator DFO can partially revert to the anti-lymphoma effect of AT 56. In FIG. 8A, AT56 inhibits the proliferation of PTCL cells, and the proliferation inhibiting effect of AT56 is partially restored after DFO is added. In fig. 8B, DFO addition partially reverts to the promotion of lipid peroxidation by AT 56. The results indicate the dependence of the iron death induction of AT56 on iron ion aggregation.

9. Fig. 9 shows that the concentration levels of iron ions in the tumor tissues of mice in different groups, the level of iron ions in the tumor tissues of the mice in the AT56 single drug group is higher than that in the blank group, and the combination of AT56 and sorafenib can significantly promote the aggregation of iron ions in the tumor tissues.

Taken together, the above studies for the first time found that PTGDS-specific inhibitor AT56 significantly inhibited the growth of PTCL, and was time-dependent and concentration-dependent. In vivo and in vitro experimental results demonstrate that AT56 promotes the iron death process by inducing iron ion aggregation in PTCL cells. Furthermore, AT56 in combination with sorafenib showed better anti-tumor effect in both PTCL cells and mouse models.

The invention can promote the deep research and clinical application of the AT56 and sorafenib combined medication scheme, and provides a solid theoretical basis and experimental basis for constructing a novel efficient combined medication strategy and optimizing a PTCL clinical diagnosis and treatment scheme.

Finally, it should be noted that: the foregoing description is only a preferred embodiment of the present invention, and the present invention is not limited thereto, but it is to be understood that modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art, although the present invention has been described in detail with reference to the foregoing embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

Application of AT56 combined with sorafenib in preparing peripheral T cell lymphoma therapeutic drugs; the molar ratio of AT56 to sorafenib is 1-20:1.
use of at56 in combination with sorafenib in any one or more of the following:

a) Preparing a product for inhibiting proliferation of peripheral T cell lymphoma cells;

b) Preparing a product for promoting iron death of peripheral T cell lymphoma cells;

c) Preparing a product for inhibiting growth of peripheral T cell lymphoma;

the molar ratio of AT56 to sorafenib is 1-20:1.
3. the use according to claim 2, wherein in b) the promotion of iron death of peripheral T cell lymphoma cells is at least manifested as promotion of iron ion aggregation in tumor tissue and/or promotion of lipid peroxidation of tumor tissue.
4. The use according to claim 2, wherein in a) -c) the product is a pharmaceutical or test product.
5. The use according to claim 4, wherein the test article is in particular a test agent for use in basic research for constructing relevant tumor cells or animal models.
6. The use according to claim 2, wherein when the product is a medicament, the medicament further comprises at least one non-pharmaceutically active ingredient.
7. The use according to claim 6, wherein the non-pharmaceutically active ingredient comprises a pharmaceutically acceptable carrier, excipient.
8. A pharmaceutical composition characterized in that the active ingredients thereof comprise AT56 and sorafenib;

the pharmaceutical composition has the effect of treating peripheral T cell lymphoma, and is specifically expressed as follows:

a) Inhibit proliferation of peripheral T cell lymphoma cells;

b) Promote iron death of peripheral T cell lymphoma cells;

c) Inhibiting peripheral T cell lymphoma growth;

the molar ratio of AT56 to sorafenib is 1-20:1.
9. the pharmaceutical composition of claim 8, further comprising at least one non-pharmaceutically active ingredient.
10. The pharmaceutical composition of claim 9, wherein the non-pharmaceutically active ingredient comprises a pharmaceutically acceptable carrier, excipient.