Application of sequential combination CAR T in preparation of antitumor drugs based on improvement of tumor microenvironment

Abstract

The invention discloses a sequential anti-tumor strategy based on combination of improved tumor microenvironment (TME) and CAR T and application of the sequential anti-tumor strategy, belongs to the technical field of biological medicine, and particularly relates to a sequential anti-tumor strategy based on combination of improved tumor microenvironment (TME) and CAR T. According to the strategy, tumor fibrosis and an acidic microenvironment are improved at the same time by applying a tumor-associated fibroblast (CAF) targeted drug and a proton pump inhibitor, and then sequential injection of CAR T is performed for combined treatment of solid tumors. Wherein the CAF targeting drug is selected from all-trans retinoic acid, calcipotriol, losartan and minetil, the proton pump inhibitor is selected from lansoprazole, omeprazole, pantoprazole, rabeprazole and esomeprazole, and the CAR T receptor or ligand is selected from mesothelin, protein tyrosine kinase 7, NKG2D, CD47, B7-H3, MUC1 and HER2; the sequential administration strategy firstly destroys a compact physical barrier of a tumor microenvironment, improves acidity of a tumor site, ensures infiltration, survival and proliferation of CAR T at the tumor site, and then utilizes specificity of CAR T to inhibit growth of solid tumors in a targeted manner, especially triple negative breast cancer with a high malignancy degree.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to an application of sequential combination of CART based on improving tumor microenvironment in the preparation of anti-tumor drugs. Specifically, tumor-associated fibroblast (CAF) targeted drugs and proton pump inhibitors synergistically improve the tumor microenvironment, followed by sequential injection of CAR T in the treatment of solid tumors. Background Art

[0002] T cells (CAR T) that have been genetically engineered to express chimeric antigen receptors on their surface have received extensive attention in the field of cancer immunotherapy. Six products have been approved by the FDA for the treatment of B-cell acute lymphoblastic leukemia and multiple myeloma, with a complete remission rate of up to 90% in patients with relapsed or refractory B-cell malignancies. However, despite the significant therapeutic effect of CAR T cells on hematological tumors, clinical data show that it is basically ineffective against solid tumors. Studies on this significant difference in efficacy have increasingly found that the tumor microenvironment (TME) associated with solid tumors has a profound impact that cannot be ignored. The tumor microenvironment is defined as the complex and rich multicellular environment in which tumors occur. Unlike hematological tumors, the TME of solid tumors presents a specific niche that is conducive to tumor progression, including physical barriers, multiple immunosuppressive mechanisms, and various biochemical factors. Therefore, CAR T therapy for solid tumors faces greater challenges. Specifically, the dense extracellular matrix, abnormal tumor vasculature, and elevated interstitial pressure in the TME hinder the infiltration of CAR T cells. In addition, nutrient deficiency and metabolic waste products (such as lactate and H) are also important factors in the development of solid tumors. + The accumulation of CAR T cells in tumor tissues forms an immunosuppressive environment that inhibits the activity of CAR T cells. Therefore, innovative technical methods or treatment strategies are urgently needed to enable CAR T cells to overcome these obstacles and improve their efficacy.

[0003] At present, the research on CAR T in solid tumors mainly focuses on selecting the best target (especially tumor-specific antigens), modifying the CAR structure, or combining immune drugs to enhance T cell response However, there is little research on improving the tumor microenvironment to enhance the infiltration, survival and proliferation of CART in the tumor site.

[0004] Tumor-associated fibroblasts (CAFs), as a hub for communication between multiple cells in the tumor stroma, are a type of stromal cell present in the interstitial tissue of tumors. When CAFs are activated by various inflammatory cytokines produced by cancer cells, immune cells, and stromal cells, CAFs rapidly proliferate and produce extracellular matrix in the enclosed tumor space, forming a physical barrier. Therefore, regulating CAFs appears to be a promising strategy to overcome the physical barriers of the TME to increase the infiltration of CAR T at the tumor site.

[0005] However, regulating a single factor is not sufficient to improve the complex TME. Acidic microenvironment, as a driver of cancer, can induce macrophage polarization to the immunosuppressive M2 phenotype, thereby promoting tumor invasion and resisting solid tumor treatment. In addition, the acidic TME induced by anaerobic metabolism of cancer cells reduces the production of cytokines, perforin, and granzyme B, thereby directly weakening the activity and function of effector T cells.

[0006] Therefore, by combining CAF-targeted drugs and proton pump inhibitors to improve tumor fibrosis and acidic microenvironment, respectively, followed by sequential injection of CAR T to exert specific anti-tumor effects, the feasibility of the treatment strategy of improving the microenvironment before anti-tumor is verified, providing a strategic reference for enhancing the efficacy of CAR T, and providing valuable insights and practical guidance for the treatment of other solid cancers based on CAR T cells, which has important clinical significance. Summary of the invention

[0007] Purpose of the invention:

[0008] The combination of CAF-targeted drugs and proton pump inhibitors can synergistically improve the fibrosis and acidity of TME, and promote the survival, proliferation and immunotherapy effects of CART cells in the tumor site.

[0009] Technical solution:

[0010] An application of sequential combination of CAR T in the preparation of anti-tumor drugs based on improving tumor microenvironment, characterized in that a CAF targeted drug or a pharmaceutically acceptable salt thereof and a proton pump inhibitor or a pharmaceutically acceptable salt thereof improve tumor fibrosis and acidic microenvironment, followed by sequential injection of CAR T for combined treatment of solid tumors.

[0011] CAF-targeted drugs bind to receptors on the nucleus of CAF cells, re-regulate the function of CAF, restore activated CAF to a quiescent state or reduce CAF activation; proton pump inhibitors bind to ATPase on tumor cells and inhibit H + The excretion of these substances improves the acidic environment at the tumor site.

[0012] At the same time, CAF-targeted drugs reduce the production of extracellular matrix and α-SMA, increasing the effectiveness of drug delivery and immunotherapy, while proton pump inhibitors can lead to acidification and accumulation of reactive oxygen species in tumor cells, hindering the proliferation and migration of cancer cells, enhancing and improving the tumor microenvironment and synergizing the anti-tumor function of CAR T.

[0013] In one aspect, the CAF targeting drug is selected from at least one of all-trans retinoic acid, calcipotriol, losartan and minnelide or a pharmaceutically acceptable salt thereof; preferably all-trans retinoic acid (ATRA).

[0014] The proton pump inhibitor is selected from one of lansoprazole, omeprazole, pantoprazole, rabeprazole and esomeprazole or a pharmaceutically acceptable salt thereof; preferably lansoprazole (LPZ).

[0015] The ligand or receptor of CAR T is selected from mesothelin, protein tyrosine kinase 7, NKG2D, CD47, B7-H3, MUC1, HER2; preferably NKG2D.

[0016] On the other hand, the single dose of the CAF targeted drug is 0.01-400 mg / kg, and preferably the single dose of all-trans retinoic acid is 0.1-50 mg / kg; more preferably 2.5 mg / kg.

[0017] The single dosage of the proton pump inhibitor is 0.01 to 500 mg / kg, and the single dosage of lansoprazole is preferably 0.1 to 100 mg / kg, and more preferably 2.5 mg / kg.

[0018] The single dose of CAR T is 1000 to 2×10 7 cells / g, preferably 1×10 4 ~2×10 6 cells / g; more preferably 1×10 5 cells / g.

[0019] The pharmaceutical composition is a CAF-targeted drug, a proton pump inhibitor and pharmaceutically acceptable excipients, and is in the form of oral liquid, granules, capsules, pills, tablets, patches or injections.

[0020] The pharmaceutically acceptable excipients are selected from one or more of diluents, excipients, fillers, binders, disintegrants, absorption enhancers, surfactants, adsorption carriers, and lubricants.

[0021] The pharmaceutical composition is preferably all-trans retinoic acid, a proton pump inhibitor, poloxamer 407, and poloxamer 188, and is in the form of an injection.

[0022] The sequential administration method is injection administration, specifically peritumoral injection of the drug composition, and intratumoral injection of NKG2D-CAR T cells after an interval of two days.

[0023] The above tumor is a solid tumor; preferably, it is triple-negative breast cancer, ovarian cancer, cervical cancer, fallopian tube cancer, prostate cancer, lung cancer, gastric cancer, liver cancer, kidney cancer, pancreatic cancer, primary peritoneal cancer or colon cancer, and most preferably, it is triple-negative breast cancer.

[0024] Specifically, the individual mechanisms of all-trans retinoic acid and lansoprazole were used to prepare poloxamer thermosensitive gel (ATRA / LPZ@gel), which was injected peritumorally to improve tumor fibrosis and acidic microenvironment. Two days later, NKG2D-CART / ATRA / LPZ was injected intratumorally to achieve targeted treatment of triple-negative breast cancer. Among them, NKG2D is frequently expressed in samples of breast cancer patients, but is expressed at a low level or not in their healthy tissues, which minimizes the clinical risk of non-tumor targeted toxicity of CAR-T therapy; the co-culture of all-trans retinoic acid and lansoprazole with NKG2D-CAR T achieved significant proliferation of CAR T and secretion of cytokines, and promoted CAR T to CD8+, which has a more tumor-killing effect. + CAR T transforms and enhances immune response.

[0025] Beneficial effects: Compared with the prior art, the present invention has the following advantages: (1) It can simultaneously improve the tumor microenvironment, including fibrosis, acidic and multiple niches, and synergistically enhance the function of CAR T; (2) It reshapes the tumor microenvironment at a safe drug concentration and combines CAR T to specifically target tumor cells, showing excellent safety and efficacy, and has clinical significance; (3) The present invention has low cost, is not limited to currently preferred drugs or tumor models, and has a wide range of applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is an inverted fluorescence image of the inhibitory effect of ATRA on activated CAFs in Example 1;

[0027] Figure 2 The Western blot results of LPZ inhibiting the expression of V-ATPase in Example 1; a is the WB result, and b is the statistical result;

[0028] Figure 3 The flow cytometry results of the NKG2D-CAR T proliferation experiment in Example 2; wherein a is the flow cytometry detection result, and b is the statistical result;

[0029] Figure 4 The results of cytokine determination in Example 2; a is TNF-α, b is IFN-γ, and c is IL-2;

[0030] Figure 5 The results of the combined efficacy experiment of ATRA and LPZ with NKG2D-CAR T in Example 2;

[0031] Figure 6Scanning electron microscope images of the thermosensitive gel in Example 3; a is Gel, b is ATRA / LPZ@gel

[0032] Figure 7 The release curves of ATRA (a) and LPZ (b) in the thermosensitive gel in Example 3;

[0033] Figure 8 The results of the effects of different groups of treatment on the pH of tumor tissues in Example 4;

[0034] Fig. 9 The Masson staining results of tumor tissues of different groups after treatment in Example 4;

[0035] Fig.10 The results of α-SMA expression in tumor tissues of different groups after treatment in Example 4;

[0036] Fig.11 The results of CD31 expression in tumor tissues of different groups after treatment in Example 4;

[0037] Fig.12 CD3 in Example 4 + Distribution results of NKG2D-CAR T in tumor tissues;

[0038] Fig.13 The quality results and appearance pictures of tumor tissues in different groups after treatment in Example 5;

[0039] Fig.14 The results of HE staining of tumor tissues in different groups after treatment in Example 5;

[0040] Fig.15 The results of TUNEL staining of tumor tissues in different groups after treatment in Example 5;

[0041] Fig.16 The CD4 and CD8 staining results of tumor tissues of different groups after treatment in Example 5;

[0042] Fig.17 The weight change results and liver and kidney function evaluation results of mice in Example 6; a is weight change, b is ALT, c is AST, d is BUN, and e is UA;

[0043] Fig.18 These are the HE staining results of the main organs of mice in Example 6. DETAILED DESCRIPTION

[0044] The technical solution of the present invention is further illustrated below in conjunction with the embodiments, which does not limit the present invention in any way. Without departing from the technical solution of the present invention, any modification or change made to the present invention that is easily implemented by a person of ordinary skill in the art will fall within the scope of the claims of the present invention.

[0045] NKG2D-CAR T cells were prepared according to CN113896801A.

[0046] Example 1: Mechanism of ATRA and LPZ

[0047] 1. Inhibitory effect of all-trans retinoic acid (ATRA) on activated CAFs

[0048] The human embryonic lung fibroblast cell line MRC-5 cells were co-cultured with the human triple-negative breast cancer cell line MDA-MB-231 cells to successfully obtain tumor-associated fibroblasts, namely activated MRC-5 cells. Specifically, the culture medium containing ATRA (15μM) was updated every day, and a control group was set up. After 7 days, the supernatant was aspirated, and after repeated washing, 4% paraformaldehyde was added to each well for fixation, and then 0.5% Triton x-100 was added, and then blocked with 5% BSA. The blocking solution was aspirated, and Rabbit anti-α-SMAprimary antibody was added to each well, and it was left to stand at 4°C overnight. The primary antibody solution was aspirated, and the washing was repeated. 500μL Anti-Rabbit IgG was added, and then the washing was repeated, and the inverted fluorescence microscope was used to take pictures.

[0049] Depend on Figure 1 As shown, it is proved that various growth factors and cytokines secreted by tumor cells can promote the transformation of MRC-5 into CAF and promote its proliferation; ATRA can significantly inhibit the activity of activated MRC-5 cells and the expression of α-SMA.

[0050] 2. LPZ inhibits the expression of V-ATPase

[0051] MDA-MB-231 cells in the logarithmic growth phase were incubated and treated with LPZ (10 μM) for 48 h in an acidic (pH 6.4) environment, and a control group was set up, with 3 replicates in each group. After 48 h, the supernatant was removed, and after repeated washing, the lysis buffer was added to extract the total protein, and the total protein was quantitatively analyzed using the BCA protein kit, and the expression of V-ATPase was analyzed using Western Blot, with α-tubulin as the internal reference protein.

[0052] Depend on Figure 2 As shown in Figure 5, LPZ can significantly reduce the expression of V-ATPase in MDA-MB-231 cells and reduce H +The excretion of TNF-α helps to increase the acidic extracellular pH, change the tumor metabolic environment, affect the growth cycle of tumor cells, and inhibit their proliferation.

[0053] Example 2: All-trans retinoic acid ATRA and lansoprazole LPZ enhance the NKG2D-CAR T

[0054] 1. NKG2D-CAR T cell proliferation assay

[0055] According to the instructions of the carboxyfluorescein succinimidyl ester (CFSE) kit, NGK2D-CAR T cells were stained. Subsequently, NKG2D-CAR T cells alone or NKG2D-CAR T cells and target cells were inoculated in a 12-well plate, and different concentrations of ATRA or LPZ were added and incubated for 48 hours. Three replicates were set for each group, and a negative control group (unstained NKG2D-CAR T cells) and a positive control group (untreated group) were set at the same time. After 48 hours, the culture medium was collected, centrifuged at 1000rpm for 5 minutes, resuspended with an appropriate amount of PBS, and the intracellular fluorescence intensity was detected by flow cytometry. The lower the fluorescence intensity, the more times the cells proliferated.

[0056] from Figure 3 It can be seen that when ATRA, LPZ and NGK2D-CAR T were administered simultaneously, NGK2D-CAR T cells proliferated more significantly.

[0057] 2. Cytokine Assay

[0058] NKG2D-CAR T cells and MDA-MB-231 cells were inoculated in 12-well plates, and the optimal concentrations of ATRA and LPZ were added and incubated for 48 hours, with 6 replicate wells for each concentration. After 48 hours, the culture medium was collected, centrifuged, and the supernatant was taken for detection according to the instructions of the ELISA kit. Finally, the OD value of each well was determined, and TNF-α, IFN-γ and IL-2 were quantitatively analyzed using the fitted standard curve.

[0059] from Figure 4 It can be seen that when ATRA and LPZ were co-incubated with NKG2D-CAR T, the secretion of three proinflammatory factors (TNF-α, IFN-γ, and IL-2) was significantly the highest, which further illustrates the synergistic effect of ATRA and LPZ with NKG2D-CAR T to enhance the immune response.

[0060] 3. Anti-tumor evaluation of NKG2D-CAR T cells

[0061] NGK2D-CAR T cells and MDA-MB-231 cells were inoculated in 96-well plates at a ratio of 1:2, 1:1, and 2:1, and specific concentrations of ATRA or LPZ were added to the NGK2D-CAR T cell group at the same time, and incubated together for 48 hours. Six replicate wells were set in each group, and a single NGK2D-CAR T cell group, a single target cell group, and a maximum target cell lysis group were set at the same time. 1 hour before the scheduled detection time point, LDH releaser was added to the maximum target cell lysis group, and the mixture was repeatedly blown several times to mix, and then cultured. After the scheduled time was reached, the culture plate was centrifuged. 120 μL of the supernatant from each well was taken and added to the corresponding wells of a new 96-well plate, 60 μL of LDH detection working solution was added to each well, and the absorbance value of each well was measured at 490 nm using an enzyme reader. Cytotoxicity was calculated as follows.

[0062]

[0063] Among them, OD c Represents the OD value of the group co-cultured with NGK2D-CAR T cells and target cells, OD z OD e OD o Represent the OD values ​​of the maximum target cell lysis group, the NGK2D-CAR T cell group alone, and the target cell group alone, respectively.

[0064] The results are as follows Figure 5 As shown, the presence of ATRA and LPZ can significantly increase the toxicity of NKG2D-CAR T to tumor cells.

[0065] Example 3: Preparation and characterization of ATRA / LPZ@gel

[0066] 1. Preparation of ATRA / LPZ@gel

[0067] Weigh an appropriate amount of poloxamer 407 and poloxamer 188 accurately and place them in a beaker, add PBS to mix, prepare gel by cold dissolution method, and let stand overnight at 4°C to obtain a uniform blank gel (Gel). Weigh ATRA and LPZ accurately and place them in a vial, add anhydrous ethanol to dissolve them by ultrasonic, then add blank gel, stir in an ice bath for 30 minutes, and let stand at 4°C to obtain ATRA / LPZ@gel.

[0068] 2. Characterization of ATRA / LPZ@gel

[0069] The blank gel (Gel) and the drug-loaded gel (ATRA / LPZ@gel) were freeze-dried, and the freeze-dried sample slices were taken and treated with gold. The microstructure of the gel was observed using a cold field electron scanning microscope (SEM). The drug-loaded gel was placed in an EP tube, PBS was added as the release medium, and stirred in a 37°C water bath. 100 μL of samples were taken at 1, 2, 4, 8, 12, 24, 48, 72, and 96 h, and the same volume of fresh medium was added at the same time. The peak areas of ATRA and LPZ were detected by HPLC, and the corresponding concentrations were calculated by substituting into the standard curve. Finally, the cumulative release percentage (Q%) was calculated.

[0070]

[0071] Among them, C1, C2, C i-1 , C i are the concentrations of ATRA / LPZ in the release medium at the 1st, 2nd, i-1, and i time points, respectively; V1 represents the total volume of the release medium; V2 represents the volume of the sample taken; and M represents the dosage.

[0072] like Figure 6 As shown in , the gel system has a dense network structure and can be used as an excellent drug carrier; Figure 7 As shown, within 48 hours, the release curve tends to be stable with a slight downward trend, indicating that the drug is completely released within 48 hours. At the same time, it is observed that the gel has been completely degraded at this time.

[0073] Example 4: Microenvironment improvement of ATRA / LPZ@gel

[0074] 1 × 10 5 The NSG mouse transplant tumor model was established with MDA-MB-231 cell suspension of 100 cells / g. Animal operations were in compliance with the Animal Ethics Committee of China Pharmaceutical University. When the tumor volume reached about 100 mm 3 At the time of the study, tumor-bearing mice were randomly divided into four groups and treated with normal saline (as a control group), ATRA@gel, LPZ@gel, and ATRA / LPZ@gel, respectively. All samples were administered by peritumoral injection at a concentration of 2.5 mg / kg. Two days later, the mice were euthanized and the tumor tissues were removed. The pH value of the maximum cross-sectional area of ​​the tumor was measured using a CK multifunctional skin pH meter. The tissue sections were subsequently stained with Masson staining and immunostained for α-SMA and CD31. To further evaluate the function of CAR T cells in vivo, CAR T cells were injected into the tumor for a second time two days after the first treatment, and the tumor sections were analyzed by CD3 immunostaining.

[0075] like Figure 8As shown in the figure, the maximum cross-sectional pH of the tumor in the control group was 6.44, indicating that the interior of the tumor tissue was acidic. After A / L@gel treatment, the cross-sectional pH of the tumor increased to 6.85, and the acidic microenvironment was significantly improved. Fig. 9 As shown in the figure, in the negative control group, the blue collagen content in the tumor tissue was abundant and distributed throughout the tumor tissue. After A / L@gel treatment, the collagen content in the tumor tissue decreased, destroying the dense collagen structure. Fig.10 and 11 As shown in Figure 3, after A / L@gel treatment, the expression of α-SMA in tumor tissue decreased, indicating that CAFs in the tumor area were reduced and the fibrotic microenvironment was improved. At the same time, the expression of CD31 increased, indicating that the tumor vascular system was expanded and IFP was reduced; Fig.12 As shown, CD3 marked T cells, the CD3 fluorescence expression in the ATRA / LPZ@gel+NKG2D-CAR T group not only increased, but also distributed more evenly inside the tumor, indicating that it promoted the infiltration and survival of NKG2D-CAR T in tumor tissue, facilitated the full contact between CAR T and tumor cells, and exerted an anti-tumor effect.

[0076] Example 5: Sequential anti-tumor study of ATRA / LPZ@gel and NKG2D-CAR T

[0077] The tumor volume (100 mm 3 ) tumor-bearing mice were randomly divided into the following five groups: (1) control group: peritumoral saline injection on day 1, and intratumoral saline injection on day 3; (2) ATRA / LPZ@gel group: peritumoral ATRA / LPZ@gel injection on day 1, and intratumoral saline injection on day 3; (3) NGK2D-CAR T group: peritumoral saline injection on day 1, and intratumoral NGK2D-CAR T cell injection on day 3; (4) ATRA / LPZ@gel+NGK2D-CAR T group: peritumoral ATRA / LPZ@gel injection on day 1, and intratumoral NGK2D-CAR T cell injection on day 3; (5) ATRA / LPZ@gel+NGK2D-CAR T / ATRA / LPZ group: peritumoral ATRA / LPZ@gel injection on day 1, and intratumoral NGK2D-CAR T / ATRA / LPZ injection on day 3. In the ATRA / LPZ@gel group, the concentrations of ATRA and LPZ were both 2.5 mg / kg, and the concentration of NKG2D-CAR T cells was 1×10 5 cells / g. The tumor volume and body weight of mice in each group were measured at the designated time points. After the experiment, tumor tissues were harvested and subjected to H&E staining, TUNEL staining, and CD4 and CD8 immunohistochemistry to evaluate histopathological characteristics and immune cell infiltration.

[0078] like Fig.13 , 14 As shown in Figures 15, the anti-tumor effect of ATRA / LPZ@gel alone was poor, and no obvious apoptosis occurred in tumor cells, mainly playing a role in improving the tumor microenvironment. The anti-cancer effect of ATRA / LPZ@gel+NKG2D-CAR T / ATRA / LPZ was the best, the dense structure of the tumor matrix was destroyed, and a large area of ​​apoptosis occurred in the tumor area, indicating that ATRA and LPZ also play a role in promoting the immune response of NKG2D-CAR T; Fig.16 As shown, after sequential administration of ATRA / LPZ@gel and NKG2D-CAR T / ATRA / LPZ, CD4 + NKG2D-CAR T cells were slightly reduced, but CD8 + The number of NKG2D-CAR T cells increased significantly, indicating that ATRA and LPZ promoted the transformation of NKG2D-CAR T cells to a more cytotoxic phenotype.

[0079] Example 6: In vivo safety evaluation

[0080] The body weight of mice was weighed during the experiment and the changes were recorded. The levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea nitrogen (BUN) and urea (UA) in the plasma of mice were measured after the experiment. HE staining analysis was performed on the main tissues (heart, liver, spleen, lung, and kidney) to evaluate whether each tissue was damaged.

[0081] like Fig.17 and 18 As shown, after treatment in each treatment group, there was no significant change in the weight of mice, indicating that each treatment group had no obvious toxicity to mice; there was no significant change in the levels of AST, ALT, BUN and UA, indicating that each treatment group had no damage to liver and kidney function; there was no obvious organ lesion in each experimental tissue, indicating that the drug caused relatively little damage to the body.

Claims

1. An application of a drug composition for improving tumor microenvironment in combination with CAR T cells in the preparation of an anti-tumor drug, characterized in that: The drug composition for improving tumor microenvironment includes a tumor-associated fibroblast CAF targeted drug and a proton pump inhibitor.

2. The use according to claim 1, characterized in that: The CAF targeting drug is selected from at least one of the following compounds or their pharmaceutically acceptable salts: all-trans retinoic acid, calcipotriol, losartan and minnelide; the proton pump inhibitor is selected from at least one of lansoprazole, omeprazole, pantoprazole, rabeprazole and esomeprazole or their pharmaceutically acceptable salts.

3. The use according to claim 1, characterized in that: The ligand or receptor in the CAR T cells is selected from mesothelin, protein tyrosine kinase 7, NKG2D, CD47, B7-H3, MUC1, and HER2.

4. The use according to claim 2, characterized in that: The CAF targeted drug is selected from the compound all-trans retinoic acid, and the proton pump inhibitor is selected from the compound lansoprazole.

5. The use according to claim 3, characterized in that: The ligand or receptor of the CAR T cell is NKG2D.

6. The use according to claim 1, characterized in that: The pharmaceutical composition is a CAF-targeted drug, a proton pump inhibitor and pharmaceutically acceptable excipients, and is in the form of oral liquid, granules, capsules, pills, tablets, patches or injections.

7. The use according to claim 6, characterized in that The pharmaceutically acceptable excipients are selected from one or more of diluents, excipients, fillers, binders, disintegrants, absorption promoters, surfactants, adsorption carriers, and lubricants.

8. The use according to claim 7, characterized in that: The pharmaceutical composition forms a poloxamer thermosensitive gel.

9. The use according to claim 1, characterized in that: The tumor is triple-negative breast cancer, ovarian cancer, cervical cancer, fallopian tube cancer, prostate cancer, lung cancer, stomach cancer, liver cancer, kidney cancer, pancreatic cancer, primary peritoneal cancer or colon cancer.

10. The solid tumor according to claim 9, characterized in that: The solid tumor is selected from triple-negative breast cancer.

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