Traditional Chinese medicine compound based on iPSC exosome as well as preparation method and application of traditional Chinese medicine compound
By loading lipid-soluble and hydrophilic components onto iPSC exosomes, traditional Chinese medicine complexes were prepared, solving the problems of co-loading and targeted delivery of multiple types of drugs, improving drug loading rate and drug synergy, and significantly improving the treatment effect of depression.
Patent Information
- Application Number
- CN202511901098.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-16
- Publication Date
- 2026-03-17
AI Technical Summary
Existing technologies cannot achieve efficient co-loading of multiple types of drugs (such as lipophilic traditional Chinese medicine, hydrophilic components and nucleic acid drugs), precise targeted delivery across the blood-brain barrier, and controllable release and quantitative evaluation of drug synergistic effects. In particular, in the treatment of depression, there are problems such as single drug loading, poor targeting, and unclear synergistic mechanism.
By loading lipid-soluble and hydrophilic components onto iPSC exosomes, a traditional Chinese medicine complex was prepared using ultrasound and co-incubation methods. This approach enables the simultaneous delivery of lipid-soluble and hydrophilic components, enhancing targeting and drug loading capacity, and improving delivery efficiency across the blood-brain barrier.
It achieves efficient co-loading of multiple types of drugs, improves drug loading and encapsulation rates, enhances drug targeting and synergistic effects, and significantly improves the efficacy of depression treatment.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of biological medicine, in particular to an iPSC exosome-based traditional Chinese medicine compound and a preparation method and application thereof. BACKGROUND
[0002] At present, the treatment of depression mainly relies on single drug delivery systems, such as chemical synthetic drugs or natural drugs alone. In recent years, exosomes have been explored as drug delivery tools due to their natural nanocarrier characteristics, but they have problems such as single type of drug loading, poor targeting, and unclear synergistic mechanism. Existing researches mainly focus on the loading of single type of drug (such as small molecules or nucleic acids) in exosomes, and lack of systematic solutions for multi-drug synergistic delivery. In addition, the low blood-brain barrier penetration efficiency and insufficient targeting accuracy in the treatment of depression limit the therapeutic effect, and there is an urgent need to develop a new delivery platform with high drug loading and precise targeting.
[0003] Current common technical means include: liposome-encapsulated small molecule drugs, which can improve solubility but cannot achieve tissue-specific targeting; viral vector delivery of gene drugs, which has high transfection efficiency but has immunogenicity risk; unmodified exosome loading of single drug, which has good biocompatibility but low drug loading and no active targeting ability. Among them, the loading rate of liposome technology for lipophilic drugs can reach 15-20%, but the encapsulation efficiency for hydrophilic drugs is less than 5%; viral vectors can achieve gene transfection efficiency of more than 90%, but may cause inflammation; the drug loading capacity of natural exosomes is usually less than 10 μg / mg protein, and lacks targeted modification.
[0004] Existing technologies cannot simultaneously meet the following needs: efficient co-loading of multiple types of drugs (such as lipophilic traditional Chinese medicine, hydrophilic ingredients, and nucleic acid drugs); precise targeted delivery across the blood-brain barrier; controllable release and quantitative evaluation of drug synergistic effect. Specifically, liposomes are difficult to stably encapsulate nucleic acid drugs, viral vectors cannot be compatible with small molecule drugs, and unmodified exosomes cannot break through the blood-brain barrier and lack drug diversity. SUMMARY
[0005] In order to solve the above technical problems, the present application provides an iPSC exosome-based traditional Chinese medicine compound and a preparation method and application thereof.
[0006] In a first aspect, the present application provides an iPSC exosome-based traditional Chinese medicine compound, which is characterized by being prepared by loading liposoluble ingredients and hydrophilic ingredients on iPSC exosomes. The diameter of the iPSC exosomes is 30-150 nm, and the total drug loading capacity of the iPSC exosomes is 40-60 μg / mg protein. The liposoluble component is selected from one or more of pachymic acid, curcumin; the loading amount of the liposoluble component is 18-33 μg / mg of protein; The hydrophilic component is selected from one or more of ginsenoside Rg1, baicalin; the loading amount of the hydrophilic component is 13-30 μg / mg of protein.
[0007] Preferably, the total drug loading amount of the iPSC exosome is 45-55 μg / mg of protein.
[0008] Preferably, the loading amount of the liposoluble component is 28-32 μg / mg of protein.
[0009] Preferably, the loading amount of the hydrophilic component is 13-22 μg / mg of protein.
[0010] In a second aspect, the application provides a preparation method of the traditional Chinese medicine compound, characterized in that the method comprises the following steps: Preparation of iPSC exosome: iPSCs are cultured, and when the confluence reaches 80%, the culture medium is replaced with an enriched culture medium, and the exosomes are harvested continuously for 4 days, followed by filtration, ultrafiltration, and chromatography; The hydrophilic component is loaded into the iPSC exosome by ultrasonic method or electroporation to obtain an intermediate; The liposoluble component is loaded into the intermediate by co-incubation method or thin film hydration method to obtain the traditional Chinese medicine compound.
[0011] Preferably, the basic culture medium comprises 60-70 mg / L of L-ascorbic acid, 12-16 μg / L of sodium selenite, 530-560 mg / L of NaHCO3, 18-20 mg / L of insulin, 9-12 mg / L of transferrin, 90-110 μg / L of FGF2, and 1.5-2.5 μg / L of TGFβ1 in DMEM / F12 medium; and the enriched culture medium comprises 60-70 mg / L of L-ascorbic acid and 12-16 μg / L of sodium selenite in DMEM / F12 medium.
[0012] Preferably, the process parameters of the ultrasonic method are as follows: power, 150-250 W; working time, 2-4 s; interval time, 4-6 s; and total ultrasonic time, 10-20 min.
[0013] Preferably, the process parameters of the ultrasonic method are as follows: power, 180-220 W; working time, 2-4 s; interval time, 4-6 s; and total ultrasonic time, 13-17 min.
[0014] Preferably, the process parameters of the co-incubation method are as follows: temperature, 37℃; and incubation time, 1-2 h to obtain the intermediate.
[0015] In a third aspect, the application provides the use of the iPSC exosome-based traditional Chinese medicine compound in the preparation of a drug for treating depression.
[0016] To sum up, the technical scheme of the application has the following effects: The iPSC exosome-based traditional Chinese medicine compound provided by the application breaks through the limitation of single drug delivery, realizes the synchronous delivery of liposoluble components and hydrophilic component traditional Chinese medicine small molecules, overcomes the poor targeting of exosomes, solves the problems of single drug type, unclear synergistic mechanism and insufficient efficacy verification means of exosomes, effectively improves the drug loading rate and encapsulation efficiency of the iPSC exosome-based traditional Chinese medicine compound, and has a broad application prospect in the drug for treating depression. BRIEF DESCRIPTION OF DRAWINGS
[0017] Figure 1 Fig. 4 is a fluorescence result diagram of each organ of the mice after the traditional Chinese medicine compound was dropped into the nose of the mice in Test Example 2.
[0018] Figure 2 Fig. 5 is a result diagram of the open field test and forced swimming test after the traditional Chinese medicine compound was dropped into the nose of the depression model mice in Test Example 3. DETAILED DESCRIPTION
[0019] The application will be further described in detail below in combination with examples, comparative examples and performance test experiments, and these examples cannot be understood as limiting the scope of the application.
[0020] The composition of the base medium used in the application is: DMEM / F12 medium containing 64 mg / L of L-ascorbic acid (Sigma-Aldrich A8960), 14 μg / L of sodium selenite (Sigma-Aldrich S5261), 543 mg / L of NaHCO3, 19.4 mg / L of insulin (Puxitang I10022), 10.7 mg / L of transferrin, 100 μg / L of FGF2, and 2 μg / L of TGFβ1. The composition of the enrichment medium is: DMEM / F12 medium containing 64 mg / L of L-ascorbic acid, 14 μg / L of sodium selenite, 543 mg / L of NaHCO3, and 19.4 mg / L of insulin. Pachyman acid is from Chengdu Puruihua Science and Technology Development Co., Ltd.; Baicalin is from Henan Standard Material Research Center; Curcumin is from Chengdu Puruihua Science and Technology Development Co., Ltd.; Ginsenoside is from Shanghai Aladdin Biochem Technology Co., Ltd.
[0021] EXAMPLE
[0022] Example 1
[0023] Embodiment 1 provides an iPSC exosome-based traditional Chinese medicine compound and a preparation method thereof.
[0024] The preparation method of the iPSC exosome-based traditional Chinese medicine compound in Embodiment 1 is shown as follows.
[0025] (1) Preparation of iPSC exosome (1.1) iPSC resuscitation and subculture Take the iPSC cryopreservation tube from the liquid nitrogen tank and quickly place it in a 37°C water bath for rapid thawing for 2 min; transfer the thawed cell suspension to a centrifuge tube containing 10 mL of pre-warmed 37°C basal medium, centrifuge at 1000 rpm for 20 sec, and discard the supernatant.
[0026] Resuspend the cell pellet with 2 mL of iPSC basal medium, evenly inoculate into a culture dish containing 10 mL of basal medium, and place in a 37°C CO2 incubator for culture. Observe the cell morphology daily, and when the cell confluence reaches 80%, discard the basal medium, wash twice with sterile PBS to remove residual medium and cell metabolic waste; add 10 mL of enriched medium and continue to culture in the incubator. Starting from the first day of changing the enriched medium, collect the culture supernatant at the same time every day for 4 consecutive days. After each collection of supernatant, immediately supplement an equal amount of exosome-enriched medium to the culture dish to ensure that the cells remain in an appropriate culture environment and maintain the efficiency of exosome secretion.
[0027] (1.2) Isolation and purification of iPSC exosome Preliminary filtration to remove impurities: combine the iPSC culture supernatant collected for 4 days and transfer it to a 50 mL centrifuge tube. First, filter it through a 0.22 μm sterile filter to remove large particle impurities such as cell debris, apoptotic bodies, and microorganisms in the supernatant, and collect the filtered clear liquid. Ultrafiltration concentration: divide the filtered liquid into 100 kDa molecular weight ultrafiltration centrifuge tubes, centrifuge at 3000 x g for 15 min at 4°C. When the concentrated volume reaches 1 / 15 of the original volume, stop the centrifugation. Add sterile PBS buffer to the original volume, centrifuge at 3000 x g for 15 min, and repeat this step 2 times to remove small molecule impurities and residual proteins in the medium, obtaining the crude exosome extract.
[0028] Chromatographic purification: The SEC column was equilibrated with sterile PBS buffer at a flow rate of 0.5 mL / min until the column pressure stabilized and the baseline was flat. The crude exosome extract was loaded onto the top of the SEC column and eluted with PBS buffer at a flow rate of 0.5 mL / min, collecting 0.5 mL of eluent from each tube. The particle concentration and size distribution in each collection tube were analyzed using a nanoparticle tracking analyzer. Fractions with particle sizes in the range of 30-150 nm were collected and combined to obtain the purified iPSC exosome suspension, which was stored at 4°C.
[0029] (2) Ultrasonic loading of hydrophilic components: Take the purified iPSC exosome suspension with a protein content of 10 μg / mL (detected by BCA method), add 2 μL of ginsenoside Rg1 hydrophilic component drug stock solution to 10 mL of exosome suspension (containing 100 μg of exosome protein) (the amount added is calculated according to the mass ratio of exosome to drug of 10:1, and the amount of drug added is 10 μg); wherein, the hydrophilic component drug stock solution is prepared by diluting with DMSO to a drug concentration of 5 mg / mL.
[0030] The mixture was transferred to sterile EP tubes and placed in an ice bath. Ultrasonic treatment was then performed using an ultrasonic homogenizer: power set to 200W, working time 3 seconds, interval time 5 seconds, total ultrasonic time 12 minutes. Temperature increases during ultrasonication were avoided to prevent exosome rupture or drug inactivation. The intermediate was obtained after ultrasonication.
[0031] (3) Co-incubation method for loading lipid-soluble components: Add 2 μL of pamoate lipid-soluble component drug stock solution to the above intermediate (the amount added is calculated according to the mass ratio of exosomes to drug of 10:1, and the amount of drug added is 10 μg); wherein, the lipid-soluble component drug stock solution is prepared by diluting with DMSO to a drug concentration of 5 mg / mL.
[0032] The mixture was gently blown and stirred until homogeneous, then placed in a constant temperature shaker at 37°C and 50 rpm and incubated in the dark for 2 hours to obtain the herbal compound.
[0033] Examples 2-4 Examples 2-4 provide a traditional Chinese medicine complex based on iPSC exosomes and its preparation method.
[0034] In Example 2: In step (2), baicalin was used to replace ginsenoside Rg1 in equal amounts.
[0035] In Example 3: In step (3), an equal amount of curcumin was used to replace pachymic acid.
[0036] In Example 4: The process parameters for the ultrasonic method in step (2) are: power 150W, working time 3s, interval time 5s, and total ultrasonic time 20min.
[0037] In Example 5: The process parameters for the ultrasonic method in step (2) are: power 360W, working time 3s, interval time 5s, and total ultrasonic time 10min.
[0038] The specific differences between the above embodiments and Embodiment 1 are as follows:
[0039] Comparative Example Comparative Example 1 Comparative Example 1 provides a traditional Chinese medicine complex based on iPSC exosomes and its preparation method.
[0040] The preparation method of the traditional Chinese medicine complex based on iPSC exosomes in Comparative Example 1 is shown below.
[0041] (1) Preparation of iPSC exosomes The method is the same as in Example 1.
[0042] (2) Co-incubation method for loading lipid-soluble components and hydrophilic components Add 2 μL of pamoate lipid-soluble drug stock solution (calculated based on an exosome to drug mass ratio of 10:1, with the drug amount being 10 μg) and 2 μL of ginsenoside Rg1 hydrophilic drug stock solution (calculated based on an exosome to drug mass ratio of 10:1, with the drug amount being 10 μg) to 10 mL of exosome suspension (containing 100 μg of exosome to drug mass ratio). The lipid-soluble drug stock solution was prepared by diluting with DMSO to a drug concentration of 5 mg / mL, and the hydrophilic drug stock solution was prepared by diluting with DMSO to a drug concentration of 5 mg / mL.
[0043] The mixture was gently blown and stirred until homogeneous, then placed in a constant temperature shaker at 37°C and 50 rpm and incubated in the dark for 2 hours. After incubation, the herbal compound was obtained.
[0044] Comparative Example 2 Comparative Example 2 provides a traditional Chinese medicine complex based on iPSC exosomes and its preparation method.
[0045] The preparation method of the traditional Chinese medicine complex based on iPSC exosomes in Comparative Example 2 is shown below.
[0046] (1) Preparation of iPSC exosomes The method is the same as in Example 1.
[0047] (2) Loading lipid-soluble components by ultrasound: Add 2 μL of pamoate lipid-soluble component drug stock solution to 10 mL of exosome suspension (containing 100 μg of exosome protein) (the amount added is calculated according to the mass ratio of exosome to drug of 10:1, and the amount of drug added is 10 μg); wherein, the lipid-soluble component drug stock solution is prepared by diluting with DMSO to a drug concentration of 5 mg / mL.
[0048] The mixture was transferred to sterile EP tubes and placed in an ice bath. Ultrasonic treatment was then performed using an ultrasonic homogenizer: power set to 200W, working time 3 seconds, interval time 5 seconds, total ultrasonic time 12 minutes. Temperature increases during ultrasonication were avoided to prevent exosome rupture or drug inactivation. The intermediate was obtained after ultrasonication.
[0049] (3) Co-incubation method for loading hydrophilic components: Add 2 μL of ginsenoside Rg1 hydrophilic component drug stock solution to the above intermediate (the amount added is calculated according to the mass ratio of exosomes to drug of 10:1, and the amount of drug added is 10 μg); wherein, the hydrophilic component drug stock solution is prepared by diluting with DMSO to a drug concentration of 5 mg / mL.
[0050] The mixture was gently blown and stirred until homogeneous, then placed in a constant temperature shaker at 37°C and 50 rpm and incubated in the dark for 2 hours to obtain the intermediate of the traditional Chinese medicine complex.
[0051] Performance testing Experimental Example 1 The drug loading and encapsulation efficiency of the traditional Chinese medicine complex were analyzed by HPLC using a C18 column.
[0052] By ultracentrifugation, 10 mL of the traditional Chinese medicine complex suspension was ultracentrifuged at 4℃ and 100,000×g for 70 min. The exosomes (containing encapsulated drugs) precipitated to the bottom of the tube. The supernatant was removed, and the exosome precipitate was resuspended in PBS. The mixture was then ultracentrifuged at 4℃ and 100,000×g for 70 min. The supernatant was removed, and the precipitate was resuspended in 500 μL of PBS. The drug concentration (C1) was then determined.
[0053] Simultaneously, the drug concentration (C2) in the untreated Chinese herbal compound suspension was detected.
[0054] Encapsulation efficiency EE (%) = C1 / C2 × 100%; Drug loading (μg / mg protein) = (exosomal protein content in C1 / 10mL traditional Chinese medicine complex suspension) × 100%; Test results are shown in Table 1.
[0055] Table 1. Detection results in the examples and comparative examples.
[0056] Experimental Example 2 Nasal drops were administered to monitor drug distribution in the brain and systemic exposure.
[0057] (1) DiR-labeled traditional Chinese medicine complex: The traditional Chinese medicine complex was incubated with DiR fluorescent dye (final concentration 5 μM) in the dark for 30 min, and the free dye was removed by column chromatography.
[0058] (2) Animal grouping: C57BL / 6 mice (SPF grade, 6-8 weeks old) were randomly divided into time point groups (0.5h, 1h, 2h, 4h, 8h, 24h, 72h), with 3 mice in each group.
[0059] (3) Nasal administration: tilt the mouse head 45° and slowly drip DiR-labeled Chinese medicine compound into each nostril to avoid choking.
[0060] (4) In vivo imaging detection: At each time point, the whole body of the mouse was scanned using a small animal in vivo imaging instrument (excitation wavelength 748nm, emission wavelength 780nm) to record the intensity and distribution area of fluorescence signals in the brain and the whole body.
[0061] (5) Organ collection and imaging observation: After in vivo imaging, the mice were euthanized, and the main organs such as brain, heart, liver, spleen, lung, kidney, and small intestine were aseptically collected. The residual blood was rinsed with physiological saline and the moisture was absorbed with filter paper. The organs were placed in the same position, and the fluorescence intensity of each gram of tissue was detected and quantitatively analyzed using an in vivo imaging system (ROI area setting).
[0062] Test results: such as Figure 1 The image shows the fluorescence results of various organs after administering the traditional Chinese medicine compound via nasal drops to mice. According to the fluorescence results of various organs in mice, the traditional Chinese medicine compound mainly appeared in the brain, lungs, stomach and intestines. No fluorescence was observed in metabolic organs such as the liver and kidneys. This indicates that nasal administration can improve the efficiency of drug delivery to the target site, reduce the liver toxicity of the drug, and improve safety.
[0063] Experimental Example 3 Mice were given LPS intraperitoneally to simulate depression, and the efficacy of the drug was verified by nasal drops of a traditional Chinese medicine compound.
[0064] (1) Animal grouping: C57BL / 6 mice (SPF grade, 6-8 weeks old) were divided into control group (Con), model group (LPS), model group Chinese medicine compound treatment group (LPS+Exo+Tcm), and model group iPSC exosome treatment group (LPS+Exo), with 3 mice in each group.
[0065] (2) Preparation of depression model: Mice in the model group, the model group treated with traditional Chinese medicine compound, and the model group treated with iPSC exosomes alone were injected intraperitoneally with 0.5 mg / kg of LPS daily for 5 consecutive days. Mice in the control group were injected intraperitoneally with an equal amount of PBS daily for 5 consecutive days.
[0066] (3) Nasal administration: Starting from day 6, administer 10 μL of PBS to the control group and model group each time, 10 μL of the drug to the traditional Chinese medicine complex treatment group in the model group each time, and 10 μL of exosomes (purified iPSC exosome suspension prepared in Example 1) to the iPSC exosome treatment group alone in the model group each time; administer continuously for 7 days.
[0067] (4) Behavioral testing: After the administration of the drug, the open field test and the forced swimming test were performed to detect the depressive state of the mice.
[0068] The open field test is used to assess an animal's spontaneous activity, exploratory behavior, and anxiety-like behavior. Results such as total distance traveled, total distance traveled in the central region, time spent in the central region, and total number of entries into the central region are measured. Figure 2 As shown.
[0069] As shown in the figure, compared with the control group (Con), the total distance of movement in the model group (LPS) was significantly reduced (P<0.001); while the total distance of movement was significantly increased after treatment with the traditional Chinese medicine compound treatment group (LPS+Exo+TCM) and the iPSC exosome treatment group alone (LPS+Exo) in the model group, and the total distance of movement after treatment with the traditional Chinese medicine compound treatment group was higher than that after treatment with iPSC exosome alone.
[0070] Compared with the control group, the model group showed a significant decrease in total central region distance, central region residence time, and total number of central region entries. However, after treatment with the traditional Chinese medicine compound treatment group and the iPSC exosome treatment group alone, the total central region distance, central region residence time, and total number of central region entries were all significantly increased. Furthermore, the total central region distance, central region residence time, and total number of central region entries were higher in the traditional Chinese medicine compound treatment group than in the iPSC exosome treatment group alone.
[0071] The forced swimming test is used to assess depressive-like behavior; the results of the abandonment time test are as follows: Figure 2 As shown in the figure, the abandonment time in the model group was significantly prolonged compared to the control group; however, the abandonment time was significantly shortened in both the traditional Chinese medicine compound treatment group and the iPSC exosome treatment group alone, with the abandonment time after treatment with the traditional Chinese medicine compound being shorter than that after treatment with iPSC exosomes alone.
[0072] The above experimental results show that LPS modeling significantly induces behavioral abnormalities in animals, including reduced spontaneous activity, increased anxiety-like behavior, and exacerbated depressive-like behavior. Both exosomes (Exo) and traditional Chinese medicine complex (Exo+TCM) can reverse these abnormalities, and the improvement effect of the traditional Chinese medicine complex is better than that of exosome treatment alone.
[0073] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.
Claims
1. An iPSC exosome-based traditional Chinese medicine compound, characterized in that, Prepared by loading the liposoluble component and the hydrophilic component into iPSC exosomes; The iPSC exosomes have a diameter of 30-150 nm, and a total drug loading capacity of 40-60 μg / mg of protein. The liposoluble component is selected from one or more of pachymic acid and curcumin, and the loading amount of the liposoluble component is 18-33 μg / mg of protein. The hydrophilic component is selected from one or more of ginsenoside Rg1 and baicalin, and the loading amount of the hydrophilic component is 13-30 μg / mg of protein.
2. The traditional Chinese medicine compound based on iPSC exosomes according to claim 1, characterized in that, The iPSC exosomes have a total drug loading capacity of 45-55 μg / mg of protein. 3.The traditional Chinese medicine compound based on iPSC exosomes of claim 1, characterized in that, The loading amount of the liposoluble component is 28-32 μg / mg of protein. 4.The traditional Chinese medicine compound based on iPSC exosomes of claim 1, characterized in that, The loading amount of the hydrophilic component is 13-22 μg / mg of protein.
5. A method for preparing the traditional Chinese medicine compound according to any one of claims 1-4, characterized in that, The method comprises the following steps: Preparation of iPSC exosomes: iPSCs are cultured in a basic medium, and when the confluence reaches 80%, the medium is replaced with an enriched medium, and exosomes are harvested by filtration, ultrafiltration and chromatography for 4 consecutive days; The hydrophilic component is loaded into the iPSC exosomes by ultrasonic method or electroporation to obtain an intermediate; The liposoluble component is loaded into the intermediate by co-incubation method or thin film hydration method to obtain the traditional Chinese medicine compound.
6. The preparation method of claim 5, characterized in that, The basic medium comprises 60-70 mg / L of L-ascorbic acid, 12-16 μg / L of sodium selenite, 530-560 mg / L of NaHCO3, 18-20 mg / L of insulin, 9-12 mg / L of transferrin, 90-110 μg / L of FGF2 and 1.5-2.5 μg / L of TGFβ1 in DMEM / F12 medium; and the enriched medium comprises 60-70 mg / L of L-ascorbic acid and 12-16 μg / L of sodium selenite in DMEM / F12 medium.
7. The preparation method of claim 5, characterized in that, The ultrasonic method has the following process parameters: power 150-250 W, working time 2-4 s, interval time 4-6 s, and total ultrasonic time 10-20 min.
8. The preparation method of claim 7, characterized in that, The ultrasonic method has the following process parameters: power 180-220 W, working time 2-4 s, interval time 4-6 s, and total ultrasonic time 13-17 min.
9. The preparation method of claim 5, characterized in that, The co-incubation method has the following process parameters: temperature 37℃, and incubation time 1-2 h.
10. Use of the traditional Chinese medicine compound based on iPSC exosomes according to any one of claims 1-4 in the preparation of a medicament for treating depression.