Application of TRPV1 receptor agonist or inhibitor and low-temperature protective agent

Abstract

The invention relates to application of a TRPV1 receptor agonist or inhibitor and a low-temperature protective agent. Compared with the prior art, the TRPV1 receptor stimulant or inhibitor is added into the preserving fluid, so that the damage problem of cells and tissues in the low-temperature preservation process of 4 DEG C is remarkably reduced, and the TRPV1 receptor stimulant or inhibitor can protect the integrity of cell membranes of various cells and reduce lysis caused by low-temperature preservation; the protection performance of the preserving fluid is improved, and the low-temperature storage life of cells and tissues is prolonged; and the TRPV1 receptor agonists such as capsaicin are natural compounds, have low toxicity and are suitable for wide application. The method is easy to operate, raw materials are easy to obtain, and preparation and use in various common biology laboratories and clinics are facilitated.

Description

Technical Field

[0001] The present invention generally relates to the field of pharmaceutical technology, and in particular to the use of a TRPV1 (Transient Receptor Potential Vanilloid 1) receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotope-labeled substance, hydrate, solvate or pharmaceutically acceptable salt thereof, and a cryoprotectant. Background Art

[0002] Cryopreservation of cells or tissues is a key technology in modern medicine, biological research and biotechnology. It can effectively prolong the storage time of cells and tissues and maintain their biological activity. It is widely used in fields such as organ transplantation and blood storage. However, problems such as cell lysis, death or loss of function often occur during cryopreservation, which seriously affects the preservation quality and clinical application effect. Therefore, optimizing cryopreservation technology and improving the survival rate of cells and tissues have important scientific significance and practical application value. It is of great significance to develop new cryoprotectants that have good biocompatibility, high preservation efficiency and high safety. Summary of the invention

[0003] Based on this, the present application provides the use of a TRPV1 receptor agonist or inhibitor, or its analogs, geometric isomers, tautomers, isotope labels, hydrates, solvates or pharmaceutically acceptable salts in the preparation of a cryoprotectant.

[0004] On the other hand, the present application also provides the use of a TRPV1 receptor agonist or inhibitor, or its analogs, geometric isomers, tautomers, isotope labels, hydrates, solvates or pharmaceutically acceptable salts in the preparation of a composition for preventing and / or reducing cell or tissue cryopreservation damage.

[0005] On the other hand, the present application also provides a cryoprotectant, which comprises a buffered salt solution for maintaining normal pH and osmotic pressure of tissues or cells and about 1-150 µM of a TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt thereof.

[0006] In another aspect, the present application also provides a method for preventing and / or reducing cell or tissue cryopreservation damage, comprising adding the cryoprotectant described herein to the external environment of the cell or tissue.

[0007] The present application found that TRPV1 receptor agonists or inhibitors can improve the survival rate of various cells at low temperatures and protect cell morphology. They can reduce the hemolysis rate and lipid peroxidation level of red blood cells during storage; and improve the cell state of skin cryopreservation. The present application proposes that TRPV1 receptor agonists or inhibitors can be used as additives for cell and tissue cryoprotectants, have a wide range of applications, and have a cryoprotective effect on various cell lines. They have high clinical value.

[0008] Compared with the prior art, this application significantly reduces the damage problem of cells and tissues during 4°C low-temperature storage by adding TRPV1 receptor agonists or inhibitors to the preservation solution. TRPV1 receptor agonists or inhibitors can protect the cell membrane integrity of various cells and reduce the lysis caused by low-temperature storage; improve the protective performance of the preservation solution and extend the low-temperature storage period of cells and tissues; and capsaicin and other TRPV1 receptor agonists are natural compounds with low toxicity and are suitable for wide application. This application is simple to operate, and the raw materials are easily available, which is convenient for preparation and use in various general biological laboratories and clinics.

[0009] Other features and advantages of the present application will be described in the following description, and partly become apparent from the description, or be understood by implementing the present application. Other advantages of the present application can be realized and obtained by the schemes described in the description and the drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The accompanying drawings are used to provide an understanding of the technical solution of the present application and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the technical solution of the present application and do not constitute a limitation on the technical solution of the present application.

[0011] Figure 1 It was shown that capsaicin increased the survival rate of LO2 cells at a low temperature of 4°C.

[0012] Figure 2 It was shown that capsaicin increased the survival rate of SY5Y cells at a low temperature of 4°C.

[0013] Figure 3 It was shown that Lorividin increased the survival rate of LO2 cells at a low temperature of 4°C.

[0014] Figure 4 It was shown that Lorividin increased the survival rate of BV2 cells at a low temperature of 4°C.

[0015] Figure 5 It was shown that Lorividin increased the survival rate of 293T cells at 4°C.

[0016] Figure 6 It was shown that capsaicin increased the survival rate of LO2 cells at a low temperature of 4°C.

[0017] Figure 7 It was shown that capsaicin and lorividin protected the cell morphology of BJ cells and K562 cells at low temperature and increased cell survival rate.

[0018] Figure 8 It was shown that capsaicin increased the survival rate of mouse ear skin cells at a low temperature of 4°C.

[0019] Fig. 9 It was shown that Lorividin reduced the hemolysis rate of erythrocytes at a low temperature of 4°C.

[0020] Fig.10 It was shown that capsaicin reduces the hemolysis rate of erythrocytes at a low temperature of 4°C.

[0021] Fig.11 The figure shows the morphology of red blood cells protected by capsaicin when stored at 4°C.

[0022] Fig.12 It was shown that capsaicin reduces the lipid peroxidation level of erythrocytes stored at 4°C. DETAILED DESCRIPTION

[0023] Unless otherwise stated, the technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the application belongs. When a certain amount, concentration or other value or parameter is expressed in the form of a range, a preferred range, or a preferred upper numerical limit and a preferred lower numerical limit, it should be understood that it is equivalent to specifically revealing any range by combining any pair of upper range limits or preferred numerical values ​​with any lower range limit or preferred numerical value, regardless of whether the range is specifically disclosed. Unless otherwise stated, the numerical ranges listed herein are intended to include the endpoints of the range and all integers and fractions (decimals) within the range.

[0024] The terms "about" and "approximately" when used with a numerical variable generally refer to the value of that variable and all values ​​of that variable are within experimental error (e.g., within a 95% confidence interval of the mean) or within ±10% of the specified value, or a wider range.

[0025] The expression "comprising" or its synonymous similar expressions "including", "containing" and "having" etc. are open-ended and do not exclude additional unrecited elements, steps or ingredients. The expression "consisting of excludes any element, step or ingredient not specified. The expression "consisting essentially of means that the scope is limited to the specified elements, steps or ingredients, plus the optional elements, steps or ingredients that do not materially affect the basic and novel characteristics of the claimed subject matter. It should be understood that the expression "comprising" encompasses the expressions "consisting essentially of" and "consisting of".

[0026] The expression "at least one" or "one or more" means 1, 2, 3, 4, 5, 6, 7, 8, 9 or more.

[0027] Currently, most of the existing preservation solutions are basic solutions, lacking effective protective additives, and unable to fully inhibit cell lysis and functional degradation. Generally, the preservation effect can only be maintained for a few hours to a few days, and it is difficult to meet the needs of longer preservation. Some protective agents may cause toxicity or side effects to cells or tissues, further limiting their application.

[0028] Capsaicin, a TRPV1 receptor agonist, is the active ingredient of chili peppers. Its chemical name is trans-8-methyl-N-vanillyl-6-nonenamide, and its molecular formula is C 18 H 27 NO 3 , the molecular weight is 305.412, and its chemical structure is shown below: .

[0029] In recent years, studies have found that capsaicin has a wide range of pharmacological effects, including antioxidant, anti-inflammatory, and anti-tumor activities. In terms of drug application, capsaicin is mainly used as an external analgesic in ointments and patches to relieve pain caused by arthritis, back pain, and neuropathy (such as postherpetic neuralgia). Among them, high-dose capsaicin patches (8%) are effective in relieving postherpetic neuralgia, HIV neuropathy, and diabetic neuropathy. However, the potential role of capsaicin in cryopreservation has not been studied.

[0030] We experimentally investigated the protective effects of TRPV1 receptor agonists or inhibitors on cells and tissues during cryopreservation at 4°C and found that they could significantly reduce the cell lysis rate and tissue cell death rate during the storage process. Subsequently, we verified the protective effect of the TRPV1 receptor agonist capsaicin in a mouse skin tissue cryopreservation model. Mouse skin tissue samples were placed in a preservation solution containing a 100 µM concentration of the TRPV1 receptor agonist capsaicin. After being stored at 4°C for 20 hours, tissue sections and immunohistochemistry revealed that the cell death rate in the TRPV1 receptor agonist capsaicin-treated group was significantly lower than that in the control group, and the integrity of the tissue structure was significantly improved.

[0031] In cryopreservation models of various cell lines (such as 293T, LO2, BV2, BJ, and K562), we used culture medium containing TRPV1 receptor agonists or inhibitors to perform cryopreservation experiments, and detected cell survival rates after the preservation. The results showed that the best effect was achieved when the concentration of TRPV1 receptor agonists or inhibitors was 10-100µM, and the cell survival rate was significantly higher than that of the control group.

[0032] Through the above experiments, we have confirmed the important role of TRPV1 receptor agonists or inhibitors in the cryopreservation of cells and tissues. TRPV1 receptor agonists or inhibitors can not only reduce the mortality of red blood cells and tissue cells, but also significantly increase the activity of cells after preservation. Compared with existing preservation solutions, TRPV1 receptor agonists or inhibitors are safe and effective additives that can significantly optimize cryopreservation technology and provide new solutions for cell and tissue preservation.

[0033] Therefore, in one aspect, the present application provides the use of a TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotope-labeled substance, hydrate, solvate or pharmaceutically acceptable salt thereof in the preparation of a cryoprotectant.

[0034] In some embodiments, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt in the cryoprotectant is about 1-150µM. In some embodiments, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt in the cryoprotectant is about 1µM, 2µM, 3µM, 4µM, 5µM, 6µM, 7µM, 8µM, 9µM, 10µM, 20µM, 30µM, 40µM, 50µM, 60µM, 70µM, 80µM, 90µM, 100µM, 110µM, 120µM, 130µM, 140µM or 150µM. In some embodiments, the concentration of the TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotopically labeled, hydrate, solvate or pharmaceutically acceptable salt thereof in the cryoprotectant is about 10-100 μM.

[0035] On the other hand, the present application also provides the use of a TRPV1 receptor agonist or inhibitor, or its analogs, geometric isomers, tautomers, isotope labels, hydrates, solvates or pharmaceutically acceptable salts in the preparation of a composition for preventing and / or reducing cell or tissue cryopreservation damage.

[0036] In some embodiments, in the composition, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt is about 1-150µM. In some embodiments, in the composition, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt is about 1µM, 2µM, 3µM, 4µM, 5µM, 6µM, 7µM, 8µM, 9µM, 10µM, 20µM, 30µM, 40µM, 50µM, 60µM, 70µM, 80µM, 90µM, 100µM, 110µM, 120µM, 130µM, 140µM or 150µM. In some embodiments, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotopically labeled, hydrate, solvate or pharmaceutically acceptable salt in the composition is about 10-100 μM.

[0037] On the other hand, the present application also provides a cryoprotectant, which comprises a buffered salt solution for maintaining normal pH and osmotic pressure of tissues or cells and about 1-150 µM of a TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt thereof.

[0038] In some embodiments, the cryoprotectant comprises about 1µM, 2µM, 3µM, 4µM, 5µM, 6µM, 7µM, 8µM, 9µM, 10µM, 20µM, 30µM, 40µM, 50µM, 60µM, 70µM, 80µM, 90µM, 100µM, 110µM, 120µM, 130µM, 140µM or 150µM of a TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotopically labeled, hydrate, solvate or pharmaceutically acceptable salt thereof. In some embodiments, the cryoprotectant comprises about 10-100µM of a TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotopically labeled, hydrate, solvate or pharmaceutically acceptable salt thereof.

[0039] In some embodiments, the buffered salt solution is selected from one or more of phosphate buffered saline (PBS) solution, Hank's balanced salt (HBS) solution, Dulbecco's modified eagle medium (DMEM), Alsever's solution, University of Wisconsin (UW) solution, and histidine-tryptophan-ketoglutarate (HTK) solution.

[0040] In some embodiments, the cryoprotectants described herein can be used for cryopreservation of cells, tissues, and organs.

[0041] In some embodiments, the TRPV1 receptor agonist is capsaicin or lorivid. In some embodiments, the TRPV1 receptor inhibitor is capsaicin.

[0042] As known to those skilled in the art, Nonivamide is a synthetic capsaicinoid, an agonist of TRPV1 receptor, and its chemical structure is shown in the following formula: .

[0043] Capsazepine is a selective TRPV1 receptor inhibitor, and its chemical structure is shown below: .

[0044] In some embodiments, other capsaicin analogs having a similar skeleton structure to capsaicin may also be used.

[0045] In another aspect, the present application also provides a method for preventing and / or reducing cell or tissue cryopreservation damage, comprising adding the cryoprotectant described herein to the external environment of the cell or tissue.

[0046] This application proves through experiments that the use of TRPV1 receptor agonists or inhibitors (such as natural compounds capsaicin, lorividin and capsaicin) at a concentration of 10μM or 100μM can improve the survival rate of various cells at low temperatures and protect cell morphology. It can reduce the hemolysis rate and lipid peroxidation level of red blood cells during storage; it can improve the cell state of skin stored at low temperatures.

[0047] The present application describes multiple embodiments, but the description is exemplary rather than restrictive, and it is obvious to those skilled in the art that there may be more embodiments and implementations within the scope of the embodiments described in the present application. Although many possible feature combinations are shown in the drawings and discussed in the specific embodiments, many other combinations of the disclosed features are also possible. Unless specifically limited, any feature or element of any embodiment may be used in combination with any other feature or element in any other embodiment, or may replace any other feature or element in any other embodiment.

[0048] The present application includes and contemplates combinations of features and elements known to those of ordinary skill in the art. The embodiments, features and elements disclosed in the present application may also be combined with any conventional features or elements to form a unique invention scheme. Any features or elements of any embodiment may also be combined with features or elements from other invention schemes to form another unique invention scheme. Therefore, it should be understood that any feature shown and / or discussed in the present application may be implemented individually or in any appropriate combination. Therefore, except for the limitations made according to the attached claims and their equivalents, the embodiments are not subject to other restrictions. In addition, various modifications and changes may be made within the scope of protection of the attached claims.

[0049] In addition, when describing representative embodiments, the specification may have presented the method and / or process as a specific sequence of steps. However, to the extent that the method or process does not rely on the specific order of the steps described herein, the method or process should not be limited to the steps of the specific order described. As will be understood by those of ordinary skill in the art, other sequences of steps are also possible. Therefore, the specific sequence of the steps set forth in the specification should not be interpreted as a limitation to the claims. In addition, the claims for the method and / or process should not be limited to the steps of performing them in the order written, and those skilled in the art can easily understand that these sequences can be changed and still remain within the spirit and scope of the embodiments of the present application.

[0050] The experimental methods in the following examples without specifying specific conditions are usually measured according to national standards. The experimental materials in the following examples without specifying the source are all commercially available raw materials. The equipment used in each step in the following examples is conventional equipment. If there is no corresponding national standard, it is carried out according to the general international standards, conventional conditions, or according to the conditions recommended by the manufacturer. Unless otherwise defined or specified, all professional and scientific terms used in this application have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equal to the recorded content can be applied to the method of this application.

[0051] Example Capsaicin (or Lorivid or Capsaicin) was added to the cell culture medium, and the cells or tissues were placed in a 4°C refrigerator overnight. The ratio of live and dead cells was measured the next day to compare the survival rates of the capsaicin (or Lorivid or Capsaicin) group and the control group at low temperatures. The results were verified on cell lines of various cell sources such as 293T, LO2, BV2, BJ, and K562, or mouse skin tissues, to explore the cryoprotective effect of capsaicin (or Lorivid or Capsaicin).

[0052] Take whole blood from mice to separate red blood cells, add it to Alder's solution for storage, divide the red blood cells into two equal parts, add 1μM, 10μM or 100μM capsaicin (or lorivid or capsaicin) to one part, and add an equal volume of solvent DMSO to the other part. Store in a 4℃ refrigerator, measure the red blood cell hemolysis rate every other week to observe whether capsaicin (or lorivid or capsaicin) can reduce the red blood cell hemolysis rate.

[0053] Example 1. TRPV1 receptor agonists or inhibitors improve the survival rate of various cell lines at low temperature of 4°C Capsaicin (or Lorivid or Capsaicin) was added to DMEM cell culture medium containing 10% fetal bovine serum, and the cells were placed in a 4°C refrigerator overnight. The next day, Calcein-AM / PI live cell / dead cell double staining reagent was added to the cell culture medium, and the cells were kept at room temperature and away from light for 15 minutes. Under a fixed field of view of a fluorescence microscope, the number of green fluorescent (live cells) and red fluorescent (dead cells) cells were counted, the ratio of live and dead cells was determined, and the survival rate of the capsaicin (or Lorivid or Capsaicin) group and the control group (DMSO without capsaicin, Lorivid or Capsaicin) at low temperature was compared. It was verified on cell lines of different cell sources, including LO2, SY5Y, BV2, BJ, and K562, to explore the cryoprotective effect of capsaicin, Lorivid or Capsaicin.

[0054] The experimental results are shown in Figure 1-Figure 6 . Figure 1 It shows that capsaicin improves the survival rate of LO2 cells at a low temperature of 4°C. It can be seen from the figure that when the concentration of capsaicin is 100µM, it has a good protective effect on LO2 cells. Figure 2 It was shown that capsaicin increased the survival rate of SY5Y cells at a low temperature of 4°C. Figure 3 It shows that Lorividin improves the survival rate of LO2 cells at a low temperature of 4°C. It can be seen from the figure that when the concentration of Lorividin is 100µM, it has a good protective effect on LO2 cells. Figure 4 It shows that Lorividin improves the survival rate of BV2 cells at a low temperature of 4°C. It can be seen from the figure that when the concentration of Lorividin is 10µM or 100µM, it has a good protective effect on BV2 cells. Figure 5 It was shown that 100 μM Lorividin increased the survival rate of 293T cells at 4°C. Figure 6It is shown that capsaicin improves the survival rate of LO2 cells at a low temperature of 4°C. It can be seen from the figure that when the concentration of capsaicin is 1µM, 10µM or 100µM, it has a good protective effect on LO2 cells. Figure 7 It was shown that capsaicin and lorividin protected the cell morphology of BJ cells and K562 cells at low temperature and increased cell survival rate.

[0055] Example 2. Capsaicin preservation solution reduces the mortality rate of mouse skin tissue cells Freshly isolated dorsal skin of mouse auricle was placed in DMEM cell culture medium containing 10% fetal bovine serum with or without capsaicin (DMSO control) and stored in a refrigerator at 4°C. Subsequently, the cell death rate of mouse skin tissue was detected by histopathological hematoxylin-eosin (HE) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay (TUNEL) staining. The experimental results are shown in Figure 8 The results showed that capsaicin significantly reduced the death rate of mouse ear skin cells after being stored at 4°C for 20 hours.

[0056] Example 3. TRPV1 receptor agonist promotes the preservation of red blood cells at 4°C Cryopreservation of erythrocytes is a commonly used blood product preservation technology, but long-term storage at 4°C will significantly increase the lysis rate of erythrocytes, affecting its clinical use effect. For the first time, we explored the application of TRPV1 receptor agonists in the cryopreservation of erythrocytes. The specific method was to place the fresh C57 mouse erythrocyte suspension in Aldrich's solution containing 100µM capsaicin or lorivid. Since capsaicin or lorivid needs to be dissolved in DMSO, the control group was placed in Aldrich's solution containing an equal amount of DMSO and no capsaicin. All samples were stored at 4°C. The absorbance of the erythrocyte supernatant at 540nm was measured weekly to determine the hemolysis rate. On the 35th day after drug administration, the erythrocyte suspension was centrifuged in a 4°C centrifuge, the preservation solution was discarded, and the erythrocytes at the bottom were used to prepare scanning electron microscopy samples. The morphology of erythrocytes was detected by scanning electron microscopy (SEM). On the 21st day of erythrocyte storage, the malondialdehyde (MDA) content detection kit (Solebo, Cat. No.: BC0020) was used. Centrifuge the red blood cell suspension at 1200 rpm for 3 minutes, take 100 μL of supernatant, add 300 μL of MDA detection working solution and 100 μL of reagent 3. After the mixture is kept in a 100°C water bath for 60 minutes, it is placed in an ice bath for cooling and centrifuged at 10000g for 10 minutes at room temperature. Take the supernatant to a 96-well plate, measure the absorbance of each sample at 532nm and 600nm, and calculate the difference between OD532 and OD600.

[0057] The experimental results are shown in Figure 9-12 As shown in the figure, Fig. 9It was shown that Lorividin reduced the hemolysis rate of erythrocytes at a low temperature of 4°C. Fig.10 It was shown that capsaicin reduces the hemolysis rate of erythrocytes at a low temperature of 4°C. Fig.11 The figure shows the morphology of red blood cells protected by capsaicin when stored at 4°C. Fig.12 It was shown that capsaicin reduces the lipid peroxidation level of erythrocytes stored at 4°C.

[0058] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limitations on the present application. Ordinary technicians in this field can change, modify, replace and modify the above embodiments within the scope of the present application.

Claims

1. Use of a TRPV1 receptor agonist or inhibitor, or its analogs, geometric isomers, tautomers, isotope-labeled substances, hydrates, solvates or pharmaceutically acceptable salts in the preparation of a cryoprotectant.

2. The use according to claim 1, wherein In the cryoprotectant, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt is about 1-150 μM.

3. The use according to claim 2, wherein In the cryoprotectant, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt is about 10-100 μM.

4. Use of a TRPV1 receptor agonist or inhibitor, or its analogs, geometric isomers, tautomers, isotope-labeled substances, hydrates, solvates or pharmaceutically acceptable salts in the preparation of a composition for preventing and / or reducing damage caused by cryopreservation of cells or tissues.

5. The use according to claim 4, wherein In the composition, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt is about 1-150 μM.

6. The use according to claim 5, wherein In the composition, the concentration of the TRPV1 receptor agonist or inhibitor, or its analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt is about 10-100 μM.

7. The use according to any one of claims 1 to 6, wherein The TRPV1 receptor agonist is capsaicin or lorivid; the TRPV1 receptor inhibitor is capsaicin.

8. A cryoprotectant comprising a buffered salt solution for maintaining normal pH and osmotic pressure of tissues or cells and about 1-150 µM of a TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt thereof.

9. The cryoprotectant according to claim 8, wherein The cryoprotectant comprises about 10-100 μM of a TRPV1 receptor agonist or inhibitor, or an analog, geometric isomer, tautomer, isotope label, hydrate, solvate or pharmaceutically acceptable salt thereof.

10. The cryoprotectant according to claim 8 or 9, wherein The TRPV1 receptor agonist is capsaicin or lorivid; the TRPV1 receptor inhibitor is capsaicin.

11. The cryoprotectant according to any one of claims 8 to 10, wherein The buffered salt solution is selected from one or more of PBS solution, HBS solution, DMEM, Aldrich solution, UW solution and HTK solution.

12. A method for preventing and / or reducing cryopreservation damage of cells or tissues, comprising adding the cryoprotectant according to any one of claims 8 to 11 to the external environment of the cells or tissues.

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