Use of exosomes in preparation of a medicament for preventing and treating chemotherapy-induced alopecia

Abstract

The application provides application of stem cell exosomes in preparation of a medicament for preventing and treating post-chemotherapy alopecia. The stem cell exosomes provided by the application can change related signal pathways, promote hair follicle repair and regeneration, and shorten the hair follicle resting phase, so as to achieve the effects of strengthening hair roots, preventing alopecia and promoting hair growth, and have a remarkable curative effect on treating and preventing post-chemotherapy alopecia.

Description

Technical Field

[0001] This application relates to the field of stem cell technology, and in particular to the application of stem cell exosomes in the preparation of drugs for the prevention and treatment of hair loss after chemotherapy. Background Technology

[0002] One of the most common side effects for cancer patients undergoing chemotherapy is hair loss, occurring 1-2 weeks after treatment. This post-chemotherapy hair loss has a significant psychological impact on many patients, particularly women. Unlike androgen-induced hair loss, post-chemotherapy hair loss has no effective treatment. Currently, the only clinical approach is to use a -15°C medical ice cap to apply cold compresses to the scalp during chemotherapy to reduce blood flow to the scalp capillaries and thus decrease hair follicle cell apoptosis. However, this method is rarely used due to its cumbersome procedure and poor user experience.

[0003] Common chemotherapy drugs primarily work by blocking the cell cycle, leading to a significant proportion of hair loss. Hair growth mainly involves three phases: anagen (growth), telogen (resting), and catagen (transitional), with approximately 90% of hair follicles in the anagen phase. Chemotherapy-induced hair loss primarily affects hair follicles in the anagen phase. The main characteristic of anagen follicles is the proliferation of epithelial compartments, where hair matrix cells exhibit maximum proliferative activity in order to form the hair shaft. Chemotherapy drugs can cause a sudden cessation of mitotic activity in hair matrix cells, resulting in a reduction and narrowing of the proximal keratinized hair shaft, leading to hair duct rupture and hair loss. This hair loss is rapid and extensive (80%–90%), typically occurring within days to weeks after chemotherapy.

[0004] In recent years, there have been reports on the use of scalp cooling therapy to prevent chemotherapy-induced hair loss in cancer patients during chemotherapy. However, the effectiveness and safety of scalp cooling remain to be discussed. There are no reports on the use of exosomes for the treatment and prevention of chemotherapy-induced hair loss. Traditional applications of exosomes in the field of hair loss (e.g., patent CN115227721A) are mainly focused on androgenetic alopecia, seborrheic alopecia, and post-hair transplant relapse, and have not been used for the treatment and prevention of post-chemotherapy hair loss. Summary of the Invention

[0005] Based on this, this application provides the application of stem cell exosomes in the preparation of drugs for preventing and treating hair loss after chemotherapy, in order to address the lack of exosome-related drugs for treating hair loss after chemotherapy.

[0006] The technical solution of this application is to provide the application of stem cell exosomes in the preparation of drugs for preventing and treating hair loss after chemotherapy.

[0007] In one embodiment, the post-chemotherapy hair loss is hair loss caused by chemotherapy drugs.

[0008] In one embodiment, the drug comprises an active ingredient and pharmaceutically acceptable excipients, the active ingredient including stem cell exosomes.

[0009] In one embodiment, the stem cell exosomes are selected from at least one of embryonic stem cell exosomes and induced pluripotent stem cell exosomes.

[0010] In one embodiment, the stem cell exosomes are human stem cell exosomes or mouse stem cell exosomes.

[0011] In one embodiment, the dosage form of the drug is a tablet, capsule, granule, pill, injection, or sustained-release formulation.

[0012] In one embodiment, the drug is a tablet, and the pharmaceutically acceptable excipient is selected from at least one of diluents, binders, disintegrants, lubricants, and humectants.

[0013] In one embodiment, the diluent is selected from at least one of starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, and calcium carbonate.

[0014] And / or, the adhesive is selected from at least one of starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, and polyethylene glycol.

[0015] And / or, the disintegrant is selected from at least one of starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium, carboxymethyl starch sodium, polyoxyethylene, sorbitol, fatty acid esters and sodium dodecyl sulfonate.

[0016] And / or, the lubricant is selected from at least one of talc, silica, stearate, tartaric acid, liquid paraffin, and polyethylene glycol.

[0017] And / or, the wetting agent is selected from at least one of water, ethanol and isopropanol.

[0018] In one embodiment, the drug is an injectable preparation, and the pharmaceutically acceptable excipient is selected from at least one of solubilizers, pH adjusters, and osmotic pressure adjusters.

[0019] In one embodiment, the solubilizer is selected from at least one of ethanol, isopropanol, propylene glycol, polyethylene glycol, poloxamer, lecithin, and hydroxypropyl-β-cyclodextrin.

[0020] And / or, the pH adjuster is selected from at least one of citrate, phosphate, carbonate, acetate, hydrochloric acid and hydroxide.

[0021] And / or, the osmotic pressure regulator is selected from at least one of sodium chloride, mannitol, glucose, phosphate, citrate and acetate.

[0022] This application provides the use of stem cell exosomes in the preparation of drugs for treating hair loss after chemotherapy. The stem cell exosomes provided by this application can alter related signaling pathways, promote hair follicle repair and regeneration, and shorten the resting phase of hair follicles, thereby achieving the effects of strengthening hair roots, preventing hair loss, and promoting hair growth. It has significant efficacy in treating and preventing hair loss after chemotherapy. Attached Figure Description

[0023] Figure 1 Verification of the morphology and expression of exosomes derived from hiPSC;

[0024] Figure 2 Verification of the morphology and expression of exosomes derived from hUC-MSC;

[0025] Figure 3 A schematic diagram illustrating the successful establishment of a mouse model of hair loss induced by cyclophosphamide chemotherapy.

[0026] Figure 4 A schematic diagram illustrating the treatment strategy for exosome therapy in cases of hair loss following chemotherapy.

[0027] Figure 5 Figure showing the comparison results between the iPS exosome group and the hUC-MSC exosome group for exosome therapy for hair loss after chemotherapy;

[0028] Figure 6 A schematic diagram illustrating the strategy for preventing hair loss after exosome therapy with chemotherapy.

[0029] Figure 7 The results of iPSC exosome therapy in preventing post-chemotherapy hair loss are shown in the figure.

[0030] Figure 8 To investigate the molecular mechanism of iPSC exosomes in preventing post-chemotherapy hair loss;

[0031] Among them, hiPSC-Eoxs: iPSC exosomes; Huc-MSC-Eoxs: MSC exosomes;

[0032] concentration; diameter; density; cyclophosphamide; relative expression; CD9, CD63, Tsg101: surface markers of exosomes. Detailed Implementation

[0033] The present application will be further described in detail below with reference to the embodiments and examples. It should be understood that these embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. Furthermore, numerous specific details are set forth in the following description to provide a fuller understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0034] Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains.

[0035] the term

[0036] The terms “combinations thereof,” “any combination thereof,” and “any combination thereof” as used in this application include all suitable combinations of any two or more of the listed items.

[0037] In this application, the term "suitable" as used in "suitable combination", "suitable method", "any suitable method", etc., refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0038] In this application, terms such as "preferred," "better," "more suitable," and "ideal" are merely used to describe implementation methods or embodiments that achieve better results, and should be understood not to limit the scope of protection of this application.

[0039] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0040] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0041] Chemotherapy-induced alopecia (CIA) is a common toxic side effect of comprehensive cancer treatment. Chemotherapy drugs can cause the sudden cessation of mitotic activity in hair follicle cells, leading to a reduction and narrowing of the proximal keratinized hair shaft, resulting in hair duct rupture and hair loss. The hair loss is rapid and extensive (80%–90%), and usually occurs within days to weeks after chemotherapy.

[0042] This application provides, in one aspect, the use of stem cell exosomes in the preparation of drugs for preventing and treating hair loss after chemotherapy. The use in drugs for preventing and treating hair loss after chemotherapy refers to their application in both treating and preventing hair loss following chemotherapy.

[0043] Stem cells are a type of cell with unlimited or immortal self-renewal capacity, capable of producing at least one type of highly differentiated daughter cell. They are classified by origin into embryonic stem cells, adult stem cells, and induced pluripotent stem cells (iPS cells). Embryonic stem cells are primitive, highly undifferentiated cells that can differentiate into more than two hundred cell types under specific conditions. Adult stem cells are undifferentiated cells found in differentiated tissues; these cells are capable of self-renewal and can specialize to form the cells that make up that type of tissue. Adult stem cells are present in various tissues and organs of the body.

[0044] Currently discovered adult stem cells mainly include: hematopoietic stem cells, bone marrow mesenchymal stem cells, neural stem cells, liver stem cells, muscle satellite cells, skin epidermal stem cells, intestinal epithelial stem cells, retinal stem cells, and pancreatic stem cells. Induced pluripotent stem cells are pluripotent stem cells obtained by reprogramming terminally differentiated somatic cells through the introduction of specific transcription factors.

[0045] Exosomes are small extracellular vesicles with a diameter of 30–150 nm. They are stable, have low immunogenicity, are non-teratogenic, and can cross the blood-brain barrier. Due to their ease of cell fusion, they can release their contents into target cells, serving as engineered carriers for drugs or drug delivery systems. Almost all cell types in the body can release exosomes, which reside extensively in body fluids. Exosomes are rich in various bioactive molecules, carrying information through proteins, lipids, and nucleic acids, mediating cell communication, proliferation, and apoptosis, participating in angiogenesis, immune regulation, and fibrosis control, and playing an important role in epigenetic regulation. Exosomes are considered an important pathway for stem cells to function through paracrine effects, providing a new strategy for cell-free therapy while avoiding the risks associated with stem cell transplantation.

[0046] In this embodiment, the stem cell exosomes are embryonic stem cell exosomes. Of course, in other embodiments, the stem cell exosomes are not limited to embryonic stem cell exosomes, but can also be other types of stem cell exosomes, such as induced pluripotent stem cell exosomes.

[0047] In one specific example, the stem cell exosomes are mouse stem cell exosomes or human stem cell exosomes. Of course, in other embodiments, the stem cell exosomes can also be of other animal origins, such as dogs, cats, etc.

[0048] This application demonstrates through mouse studies that stem cell exosomes can significantly improve and prevent post-chemotherapy hair loss caused by chemotherapy drugs.

[0049] Another aspect of this application provides a medicament for treating hair loss following chemotherapy, comprising an active ingredient and pharmaceutically acceptable excipients, said active ingredient including stem cell exosomes.

[0050] An active ingredient refers to any component that provides pharmacological activity or other direct effects in the diagnosis, cure, alleviation, treatment, or prevention of disease, or that affects the structure or any function of the human or other animal body. Specifically, in this embodiment, the active ingredient includes stem cell exosomes. In one embodiment, the active ingredient is a stem cell exosome.

[0051] In one specific example, the active ingredient is at least one of embryonic stem cell exosomes and induced pluripotent stem cell exosomes.

[0052] Optionally, stem cell exosomes are not limited to embryonic stem cell exosomes and induced pluripotent stem cell exosomes mentioned above, but may also be exosomes secreted by other types of stem cells. It is understood that in some embodiments, the active ingredient is not limited to stem cell exosomes, but may also be other ingredients effective in treating chemotherapy-induced alopecia syndrome.

[0053] In a specific example, the active ingredient of the aforementioned drug is a combination of stem cell exosomes and other active ingredients.

[0054] Pharmaceutically acceptable excipients are pharmaceutically acceptable auxiliary materials or carriers that are compatible with other components of a pharmaceutical preparation and suitable for contact with the tissues or organs of the recipient (e.g., human or animal). They do not cause, or cause very minor, toxic, irritant, allergic reactions, immunogenicity, or other complications during use.

[0055] Optionally, the aforementioned drugs for post-chemotherapy hair loss may be oral preparations such as tablets, disintegrating tablets, capsules, granules, pills, and oral liquids, or injectable preparations such as injections, lyophilized powder injections, large-volume infusions, liposome injections, and microsphere injections; of course, they may be ordinary preparations, or novel or special preparations such as sustained-release preparations, controlled-release preparations, targeted preparations, nano-preparations, etc.

[0056] In a specific example, the drug for treating post-chemotherapy hair loss is a tablet. To formulate the aforementioned drug for treating post-chemotherapy hair loss into a tablet, various excipients known in the art can be used as pharmaceutically acceptable excipients. Specifically, since the drug for treating post-chemotherapy hair loss is a tablet, the pharmaceutically acceptable excipient is selected from at least one of diluents, binders, wetting agents, disintegrants, and lubricants.

[0057] Further, the diluent is selected from at least one of starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, and calcium carbonate. The humectant is selected from at least one of water, ethanol, and isopropanol. The binder is selected from at least one of starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, and polyethylene glycol. The disintegrant is selected from at least one of starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium, sodium carboxymethyl starch, polyoxyethylene, sorbitol, fatty acid ester, and sodium dodecyl sulfonate. The lubricant is selected from at least one of talc, silica, stearate, tartaric acid liquid paraffin, and polyethylene glycol. Of course, tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0058] In a specific example, the aforementioned drug for treating post-chemotherapy hair loss is a special capsule formulation such as a capsule, soft capsule, targeted capsule, or fast-dissolving capsule. To make the aforementioned drug for treating post-chemotherapy hair loss a special capsule formulation such as a capsule, soft capsule, targeted capsule, or fast-dissolving capsule, the aforementioned stem cell exosomes can be mixed with diluents, flow aids, etc., and then the resulting mixture can be directly placed into hard capsules, soft capsules, or special capsules. Of course, in some embodiments, the aforementioned stem cell exosomes can also be first mixed with at least one of a diluent, binder, and disintegrant to form particles, microspheres, microspheres, liposomes, etc., before being placed into hard capsules, soft capsules, or special capsules.

[0059] Optionally, the above-mentioned drug for treating post-chemotherapy hair loss is an injectable preparation. To prepare the above-mentioned drug for treating post-chemotherapy hair loss into an injectable preparation, water, ethanol, isopropanol, propylene glycol, polyethylene glycol, or mixtures thereof can be used as solvents, and an appropriate amount of pharmaceutically acceptable excipients commonly used in the art can be added before use.

[0060] Specifically, the aforementioned drugs for treating post-chemotherapy hair loss are injectable formulations, and pharmaceutically acceptable excipients are selected from at least one of solubilizers, pH adjusters, and osmotic pressure adjusters. Specifically, the solubilizer is selected from at least one of ethanol, isopropanol, propylene glycol, polyethylene glycol, poloxamer, lecithin, and hydroxypropyl-β-cyclodextrin. The pH adjuster is selected from at least one of citrate, phosphate, carbonate, acetate, hydrochloric acid, and hydroxide. The osmotic pressure adjuster is selected from at least one of sodium chloride, mannitol, glucose, phosphate, citrate, and acetate. If preparing a lyophilized powder for injection, mannitol, glucose, etc., may also be added as a supporting agent. Furthermore, if necessary, colorants, preservatives, fragrances, and other additives may also be added to the pharmaceutical preparation.

[0061] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0062] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0063] Example 1 Cell Culture

[0064] I. iPS cell culture:

[0065] iPSC cells were obtained from ATCC, ATCC number: ACS-1011. Pellacin was obtained from Nuwacell, catalog number: RP01002. ncTarget serum-free medium was purchased from Nuwacell, catalog number: RP01020. ROCK inhibitor was purchased from Beyotime, catalog number: SC6664.

[0066] The culture flask was coated with hydrin at a working concentration of 1 μg / cm³. 2 Incubate at room temperature (25°C) for at least 1 hour. When using, aspirate the coating solution with a pipette. After cell resuscitation, add Rock inhibitor (10mM stock solution diluted 1:4000) and mix thoroughly. Seed the cells in culture dishes at a density of 10%–20%, changing the medium daily until the cells reach 80% passage.

[0067] II. Mesenchymal stem cell culture:

[0068] Primary and passaged umbilical cord mesenchymal stem cells were cultured using mesenchymal culture medium (eMSC Medium from Yuansheng Biotechnology). The mesenchymal stem cell batch number was UC2019001, and the cells were passaged every three days.

[0069] Example 2: Preparation and Identification of Exosomes

[0070] I. Exosome preparation:

[0071] 1. Thaw the collected cell culture supernatant at 4°C and centrifuge at 500g for 5 minutes at 4°C to remove dead cells.

[0072] 2. Collect the supernatant and centrifuge at 2000g for 20 minutes at 4℃ to remove cell debris.

[0073] 3. Collect the supernatant and centrifuge at 4℃ and 10000g for 30 minutes to remove larger vesicles.

[0074] 4. Collect the supernatant and centrifuge at 100,000g for 100 minutes at 4°C to collect exosomes.

[0075] 5. The precipitate after ultracentrifugation was resuspended in 50 ml of PBS and then further purified by ultracentrifugation at 100,000 g for 100 min at 4 °C.

[0076] 6. Resuspend the exosomes in an appropriate amount of PBS, filter sterilize using a 0.22 μm filter, determine the exosome concentration using the Brand Ford method, aliquot, and store at -80℃.

[0077] II. Exosome Identification:

[0078] The following three methods are used to identify exosomes, and the specific steps include:

[0079] (1) After negative staining of exosomes, their morphology was observed using transmission electron microscopy. In the experiment, the copper mesh used for negative staining was destaticated under vacuum for 2 min. 5 μL of diluted sample was dropped onto the surface of the copper mesh, and excess sample was removed with filter paper. 4 μL of uranium acetate dye was added and stained for 1 min. The residual dye was removed with filter paper, and 4 μL of dye solution was added again. After 1 min, the solution was removed and dried at room temperature before observation under electron microscopy.

[0080] (2) Dynamic light scattering was used to analyze the particle size of exosomes. Pure water and PBS were used as controls. 10 μL of diluted sample was added to the detection tube for detection.

[0081] (3) The expression of exosome surface markers CD9, CD63 and Tsg101 was detected by Western blotting.

[0082] Depend on Figure 1 As shown in Figure A, electron microscopy imaging after negative staining reveals that exosomes exhibit a typical cup-shaped structure or a hemispherical structure with one side concave. Figure 1 As shown in Figures B to C, particle size analysis of the isolated exosomes revealed that most exons had diameters ranging from 80 to 140 nm using dynamic light scattering. Figure 1 As shown in Figure D, CD63 and Tsg101 were expressed in the collected exosomes, with CD63 highly expressed, Tsg101 moderately expressed, and GAPDH rarely or almost not expressed. Figure 2 As shown in A to D, the same results were obtained for human embryonic stem cell exosomes.

[0083] Example 3: Establishment of a cyclophosphamide-induced hair loss model in mice after chemotherapy

[0084] The growth cycle of mouse hair follicles differs slightly from that of humans. In adults, 90% of hair follicles are in the anagen phase, and chemotherapy drugs can cause hair loss during the anagen phase. However, mouse hair follicles are mainly in the telogen phase. The modeling strategy involves first removing the hair from the mice to induce the hair follicles to be in the anagen phase, and then administering chemotherapy drugs.

[0085] Modeling method: After anesthetizing C57BL / 6 mice, a 1:1 mixture of rosin and paraffin was heated and melted, then applied to the back of the mice. After solidification and hardening, the mixture was peeled off to induce hair follicles into the anagen phase and hair growth. The hair removal area for each mouse was the back. New hair was observed growing on the back of the mice after 9 days. Figure 3 As shown, hair had grown on the back of the control group after 9 days, while the back of the chemotherapy group remained pink, indicating that the model was successfully established and further experiments could be conducted.

[0086] Example 4: Exosome therapy for post-chemotherapy hair loss

[0087] An efficacy study was conducted using exosomes from multiple sources to treat chemotherapy-induced hair loss.

[0088] Treatment experimental method: Mice were treated with wax paper for hair removal. On day 9, cyclophosphamide was injected via tail vein to establish a mouse model of chemotherapy-induced alopecia (CIA). The normal control group received an equal volume of PBS solution, while all other groups received a single subcutaneous injection of cyclophosphamide 150 mg / kg to induce severe hair loss. The treatment strategy was as follows: Figure 4 As shown.

[0089] The mice were divided into the following groups: a control group (n=10), a cyclophosphamide group (n=10), an iPS exosome group (n=10), and an hUC-MSC exosome group (n=10). On days 11 and 13, mice were subcutaneously injected at multiple points on their backs with either 1 μg / μl of exosomes or 100 μl of PBS. Overall hair growth was assessed and hair follicle damage was analyzed until day 20.

[0090] The results are as follows Figure 5 As shown, the results on day 20 (D20) revealed significant hair growth in the iPSC exosome group (iPSC-ex), with grayish-black hairs appearing on the back, while almost no new hair growth was observed in the MSC exosome group (MSC-ex). Therefore, as... Figure 5 As shown in Figure A, the iPSC exosome group was significantly superior to the MSC exosome group and the control group, while there was no difference between the MSC exosome group and the control group. Furthermore, longitudinal sections of skin tissue from each group were stained with hematoxylin and eosin (HE), as shown in Figure A. Figure 5As shown in Figure B, the iPSC-ex group significantly reduced CIA-induced follicular enlargement, while the MSC-ex group showed no significant difference compared to the CIA group, consistent with the aforementioned results. Further investigation of signaling pathways related to follicular inflammation and regeneration revealed that, for example... Figure 5 As shown in C-D, both the iPSC-ex and MSC-ex groups significantly reduced inflammatory symptoms in hair follicle tissue. iPSC-ex significantly increased the expression of insulin-like growth factor (IGF), while there was no significant difference between the MSC-ex group and the Control group.

[0091] Example 5: Exosomes for the prevention of post-chemotherapy hair loss

[0092] The key difference in prevention and treatment strategies between chemotherapy-induced hair loss and other types of hair loss lies in the predictability of hair loss after chemotherapy, thus allowing for drug intervention before chemotherapy. Therefore, an experiment was designed to involve exosomes from multiple sources before chemotherapy.

[0093] Prevention Experiment Methods: On day-3 (D-3) and day-1 (D-1), mice were subcutaneously injected at multiple points on their backs with either 1 μg / μl of exosomes or 100 μl of PBS. On day 0 (D0), mice were treated with wax paper for hair removal. On day 9, cyclophosphamide was injected via the tail vein to establish a mouse model of chemotherapy-induced alopecia (CIA). Except for the normal control group, which received an equal volume of PBS solution, all other groups received a single subcutaneous injection of cyclophosphamide 150 mg / kg to induce severe hair loss. Overall hair growth was assessed and hair follicle damage was analyzed until day 20.

[0094] The mice were then divided into three groups: a control group (10 mice), a cyclophosphamide group (CIA) (10 mice), and an iPS exosome group (iPS-ex) (10 mice). The modeling procedure is as follows: Figure 6 As shown.

[0095] Prevention experiment results such as Figure 7 As shown, similar to the treatment group, the results on day 20 (D20) showed significant hair growth in the iPSC exosome group (iPSC-ex), with grayish-black hairs growing on the back. Therefore, as Figure 7 As shown in Figure A, the iPSC exosome group was significantly superior to the control group. Furthermore, longitudinal sections of skin tissue from each group were stained with hematoxylin and eosin (HE), as shown in Figure A. Figure 7 As shown in Figure B, the iPSC-ex group significantly reduced CIA-induced follicular enlargement. Compared to the treatment group, more hair grew, indicating that iPSC-ex is superior to treatment in preventing post-chemotherapy hair loss.

[0096] The off-target effects of chemotherapy are usually caused by stimulating apoptosis in normal, highly proliferating cells. Hair loss is no exception; excessive apoptosis and loss of proliferative capacity of hair matrix keratinocytes are the causes of related hair loss. Among them, p53-mediated apoptosis is an important pathogenic mechanism of chemotherapy-induced hair loss. The downstream Fas ligand activated by p53 interacts with the Fas receptor, stimulating prespase-8, which activates caspase-3 during proteolytic cleavage, leading to apoptosis. Next, p53 immunoblotting experiments were performed on the back skin tissue of mice in each group on day 20 (p53 antibody, purchased from Beyotime).

[0097] The results are as follows Figure 8 As shown in Figure A, iPSC-ex significantly reduced P53 expression in the skin of bald patches. Furthermore, KI67 (ABCAM, catalog number AB16667) staining of skin tissue sections yielded statistical results as follows: Figure 8 As shown in Figure B, the iPSC-ex prevention group exhibited significantly increased proliferative capacity. Furthermore, skin tissue sections were stained with TUNEL (ROCHE, catalog number 11966006001), and the statistical results are shown below. Figure 8 As shown in Figure C, the apoptosis level in the iPSC-ex prevention group was significantly lower than that in the CIA group.

[0098] The embodiments described above are merely illustrative of several implementation methods of this application, intended to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Furthermore, it should be understood that after reading the above teachings of this application, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the scope of protection of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. Application of stem cell exosomes in the preparation of drugs for the prevention and treatment of hair loss after chemotherapy; The hair loss mentioned above is hair loss caused by chemotherapy drugs; The drug comprises an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient includes stem cell exosomes; The stem cell exosomes are induced pluripotent stem cell exosomes; The chemotherapy drug is cyclophosphamide.

2. The application according to claim 1, characterized in that, The stem cell exosomes are human stem cell exosomes or mouse stem cell exosomes.

3. The application according to any one of claims 1 to 2, characterized in that, The dosage form of the drug is tablets, capsules, granules, pills, or injections.

4. The application according to claim 3, characterized in that, The drug is a tablet, and the pharmaceutically acceptable excipient is selected from at least one of diluents, binders, disintegrants, lubricants, and humectants.

5. The application according to claim 4, characterized in that, The diluent is selected from at least one of starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, and calcium carbonate; And / or, the adhesive is selected from at least one of starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone and polyethylene glycol; And / or, the disintegrant is selected from at least one of starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium, carboxymethyl starch sodium, polyoxyethylene, sorbitol, fatty acid ester and sodium dodecyl sulfate; And / or, the lubricant is selected from at least one of talc, silica, stearate, liquid paraffin and polyethylene glycol; And / or, the wetting agent is selected from at least one of water, ethanol and isopropanol.

6. The application according to claim 4, characterized in that, The drug is an injectable preparation, and the pharmaceutically acceptable excipients are selected from at least one of solubilizers, pH adjusters, and osmotic pressure adjusters.

7. The application according to claim 6, characterized in that, The solubilizer is selected from at least one of ethanol, isopropanol, propylene glycol, polyethylene glycol, poloxamer, lecithin, and hydroxypropyl-β-cyclodextrin; And / or, the pH adjuster is selected from at least one of citrate, phosphate, carbonate, acetate, hydrochloric acid and hydroxide; And / or, the osmotic pressure regulator is selected from at least one of sodium chloride, mannitol, glucose, phosphate, citrate and acetate.

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