Application of selenocysteine or derivative thereof in resisting chemotherapy injury

Abstract

The invention provides application of selenocysteine or a derivative thereof in resisting chemotherapy injury. Experiments prove that selenocysteine or a derivative thereof can promote proliferation and growth of hematopoietic stem cells, enhance functions of the hematopoietic stem cells, inhibit apoptosis of the hematopoietic stem cells, resist chemotherapy injury, promote recovery of hematopoietic stem cells, leukocytes and platelets after chemotherapy and prolong the lifetime after chemotherapy.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular relates to the application of selenocysteine or its derivatives in resisting chemotherapy damage. Background Art

[0002] Chemotherapy is a conventional treatment for malignant tumors, some autoimmune diseases, parasitic diseases, and other diseases. However, chemotherapy can cause a range of side effects, including bone marrow suppression, kidney disease, liver disease, gastrointestinal disease, mucosal damage, dry mouth, neurotoxicity, cardiotoxicity, ototoxicity, hair loss, and pulmonary fibrosis. The most common side effect is bone marrow suppression, which can induce anemia, thrombocytopenia, and neutropenia, leading to fatigue, increased bleeding, and an increased risk of serious infection, seriously endangering patients' lives and health.

[0003] Clinically, myelosuppression is primarily treated with symptomatic treatment to improve the decline in blood cells. Commonly used blood cell-boosting drugs include shark liver alcohol, lecithin, vitamin B12, folic acid, and iron supplements. Hematopoietic factors such as granulocyte colony-stimulating factor and erythropoietin are also used. Conventional blood cell-boosting drugs are slow-acting and poorly effective, while cytokines are more effective but have significant adverse reactions and can lead to hematopoietic stem cell depletion, resulting in unsatisfactory clinical results.

[0004] Therefore, there is an urgent need to find a safer and more effective drug to resist chemotherapy damage. Summary of the Invention

[0005] In view of this, in order to make up for the deficiencies of the prior art, the present invention is proposed.

[0006] The first aspect of the present invention provides the use of selenocysteine or its derivatives in the preparation of drugs for resisting chemotherapy damage.

[0007] In some embodiments, the selenocysteine derivative includes a compound in which an alkyl group is attached to a selenium atom.

[0008] In the present invention, the term "alkyl" includes fully saturated branched or unbranched (e.g., straight chain or linear) hydrocarbon moieties containing 1 to 20 carbon atoms. Preferably, the alkyl group contains 1 to 7 carbon atoms, more preferably 1 to 4 carbon atoms. Representative examples of alkyl moieties include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl.

[0009] In a specific embodiment, the alkyl group is selected from methyl, and the selenocysteine derivative is selenomethylselenocysteine.

[0010] In some embodiments, the selenomethylselenocysteine includes L-selenomethylselenocysteine, D-selenomethylselenocysteine, and DL-selenomethylselenocysteine.

[0011] In a specific embodiment, the selenomethylselenocysteine is L-selenomethylselenocysteine.

[0012] In some embodiments, the selenocysteine or its derivatives include selenocysteine or its derivatives themselves (hereinafter referred to as compounds), hydrates, pharmaceutically acceptable salts, solvates or crystalline forms.

[0013] In the present invention, pharmaceutically acceptable salts refer to acidic salts formed with inorganic and / or organic acids and basic salts formed with inorganic and / or organic bases. Furthermore, when the compound contains a basic moiety (such as, but not limited to, pyridine or imidazole) and an acidic moiety (such as, but not limited to, a carboxylic acid), zwitterions may form, and such zwitterions are included in the pharmaceutically acceptable salts described herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, but other salts are also useful. Pharmaceutically acceptable salts of the compounds can be formed, for example, by reacting the compound with an amount of an acid or base in a medium, such as a medium in which the salt precipitates or an aqueous medium (followed by lyophilization).

[0014] Specific pharmaceutically acceptable salts include those salts that are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reaction, etc., and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts (pharmaceutically acceptable salts) are well known in the art. Pharmaceutically acceptable salts of the compounds described herein include salts derived from suitable inorganic and organic acids, and inorganic and organic bases.

[0015] Examples of pharmaceutically acceptable nontoxic acid addition salts are salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, perchloric acid, or salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid. Salts formed using conventional methods in the art, such as ion exchange methods, are also included. Other pharmaceutically acceptable salts include: adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, gluconate, glycerophosphate, hemisulfate, heptanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, and ammonium salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium salts, and the like.

[0016] In the present invention, a hydrate refers to a compound combined with water.

[0017] In the present invention, the solvate refers to a form of a compound or salt thereof that is associated with a solvent, typically formed by a solvolysis reaction. This physical association may include hydrogen bonding. Conventional solvents include methanol, ethanol, acetic acid, DMSO, THF, diethyl ether, and the like. In some cases, the solvate can be isolated, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. Solvates include solvates in solution and isolatable solvates.

[0018] In the present invention, the crystalline form refers to the crystalline form of a compound with a specific crystal packing arrangement. Different crystalline forms generally have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystal shapes, photoelectric properties, stability, and solubility. Recrystallization solvents, crystallization rates, storage temperatures, and other factors can lead to one crystalline form dominating. Various polymorphs of a compound can be prepared by crystallization under different conditions. In the present invention, crystalline form also includes specific crystal states, such as amorphous.

[0019] In some embodiments, the chemotherapy includes, but is not limited to, administration of chemotherapeutic drugs, e.g., paclitaxel, alkylating agents such as nitrogen mustards (e.g., mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil), nitrosoureas (e.g., carmustine (BCNU), lomustine (CCNU), semustine (methyl-CCNU)), ethylenimines / methylmelamines (e.g., thriethylenemelamine (TEM), triethylene, thiophosphoramide (thiotepa), hexamethylmelamine (HMM, altretamine)), alkyl sulfonates ... sulfonates (e.g., busulfan), triazines (e.g., dacarbazine (DTIC)); antimetabolites such as folic acid analogs (e.g., methotrexate, trimetrexate), pyrimidine analogs (e.g., 5-fluorouracil, capecitabine, fluorodeoxyuracil, gemcitabine, cytosine arabinoside); arabinoside (AraC, cytarabine), 5-azacytidine, 2,2'-difluorodeoxycytidine), purine analogs (e.g., 6-mercaptopurine, 6-thioguanine, azathioprine, 2'-deoxycoformycin (pentostatin), erythrohydroxynonyladenine (EHNA), fludarabine phosphate, 2-chlorodeoxyadenosine (cladribine, 2-CdA));Antimitotic drugs developed from natural products (e.g., paclitaxel, vinca alkaloids (e.g., vinblastine (VLB), vincristine, vinorelbine), docetaxel, estramustine, estramustine phosphate), epipodophylotoxins (e.g., etoposide, teniposide), antibiotics (e.g., actimomycin D, daunomycin (rubidomycin), daunorubicon, doxorubicin, epirubicin, mitoxantrone, idarubicin, bleomycin, plicamycin (Guangshen), daunomycin (daunorubicin), daunorubicin ... mithramycin), mitomycin C, actinomycin), enzymes (e.g., L-asparaginase), biological response modifiers (e.g., interferon-α, IL-2, G-CSF, GM-CSF); miscellaneous agents, including platinum coordination complexes (e.g., cisplatin, carboplatin, oxaliplatin), anthracenediones (e.g., mitoxantrone), substituted ureas (i.e., hydroxyurea), methylhydrazine derivatives (e.g., N-methylhydrazine (MIH), procarbazine), adrenocortical suppressants (e.g., mitotane (o,p'-DDD), aminoglutethimide);Hormones, antagonists, including adrenocortical steroid antagonists (e.g., prednisone and equivalents, dexamethasone, aminoglutethimide), progestins (e.g., hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrolacetate), estrogens (e.g., diethylstilbestrol, ethinylestradiol, and equivalents); antiestrogens (e.g., tamoxifen), androgens (e.g., testosterone propionate); One or more of the following: propionate, fluoxymesterone and its equivalents), anti-androgens (e.g., flutamide, gonadotropin-releasing hormone analogs, leuprolide), and non-steroidal anti-androgens (e.g., flutamide).

[0020] In some embodiments, the chemotherapeutic drug is selected from the class of antimetabolites.

[0021] In a specific embodiment, the chemotherapeutic drug is selected from 5-fluorouracil.

[0022] In the present invention, the term "resistance" includes prevention and / or treatment. As used herein, "prevention" means starting administration before chemotherapy and hindering the onset of damage caused thereby. As used herein, "treatment" means that the subject has developed at least one damage during the chemotherapy period, and the further progression and / or symptoms of the damage are slowed and / or alleviated.

[0023] In some embodiments, examples of subjects include, but are not limited to, mammals, e.g., humans, non-human primates, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, transgenic non-human animals. In a specific embodiment, the subject is a human.

[0024] In some embodiments, the chemotherapy injury includes but is not limited to myelosuppression, renal disease, liver disease, gastrointestinal disease, mucosal damage, dry mouth, neurotoxicity, cardiotoxicity, ototoxicity, alopecia, pulmonary fibrosis.

[0025] In some embodiments, the chemotherapy insult is selected from myelosuppression.

[0026] In some embodiments, the bone marrow suppression includes, but is not limited to, a decrease or impairment of hematopoietic stem cells, red blood cells, white blood cells, or platelets.

[0027] In some embodiments, the hematopoietic stem cells include long-term hematopoietic stem cells and short-term hematopoietic stem cells.

[0028] In some embodiments, the subject is administered before, simultaneously with, and / or after chemotherapy. It is usually administered in a pharmaceutically effective amount. In some embodiments, it has been shown that daily intraperitoneal injection of 0.1-10 μg / kg, preferably daily intraperitoneal injection of 0.55 μg / kg of selenocysteine or its derivatives can effectively resist damage. The daily amount of selenocysteine or its derivatives can be administered in a single dose or in multiple divided doses, such as twice or three times, preferably in a single dose. According to one embodiment of the present invention, for example, 5 to 15 days, a single administration of 0.55 μg / kg of selenocysteine or its derivatives is administered to the subject, preferably during 10 days.

[0029] A second aspect of the present invention provides a pharmaceutical composition comprising selenocysteine or a derivative thereof, which can resist chemotherapy damage.

[0030] In some embodiments, the pharmaceutical composition further comprises other drugs with anti-chemotherapy damage effects and organ protection drugs.

[0031] In some embodiments, the other drugs with anti-chemotherapy damage effects and organ protection drugs include but are not limited to batyl alcohol, leucine, vitamin B12, folic acid, iron, hematopoietic factors, and cytokines.

[0032] In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0033] The pharmaceutically acceptable carriers and / or excipients described herein include any substance suitable for use in a subject without undue adverse side effects (e.g., toxicity, irritation, and allergic reactions), that is, any substance with a reasonable benefit / risk ratio. The pharmaceutically acceptable carriers and / or excipients are used, as needed, to enhance the stability of the formulation, improve its activity or bioavailability, or produce an acceptable taste or odor when administered orally. The pharmaceutical composition thus formulated can be administered by any suitable route known to those skilled in the art, as needed. When administering the pharmaceutical composition, a safe, suitable, and pharmaceutically effective amount of the pharmaceutical composition of the present invention is administered to the subject.

[0034] The pharmaceutical composition of the present invention may be contained in a pharmaceutically acceptable carrier and / or excipient, wherein the pharmaceutically acceptable carrier and / or excipient includes a diluent, a binder, a surfactant, a wetting agent, an adsorption carrier, a lubricant and / or a disintegrant. Among them, diluents include but are not limited to lactose, sodium chloride, glucose, urea, starch, and water; binders include but are not limited to starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, ethylcellulose, polyvinyl alcohol, polyethylene glycol, polyvinyl pyrrolidone, alginic acid and alginates, xanthan gum, hydroxypropyl cellulose and hydroxypropyl methylcellulose; surfactants include but are not limited to polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, monoglyceride of stearic acid, and cetyl alcohol; wetting agents include but are not limited to glycerol and starch; adsorption carriers include but are not limited to starch, lactose, bentonite, silica gel, kaolin, and bentonite; lubricants include but are not limited to zinc stearate, glyceride monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearyl fumarate, polyoxyethylene monostearate, monolauric sucrose ester, sodium lauryl sulfate, magnesium lauryl sulfate, and magnesium lauryl sulfate.

[0035] The appropriate dosage of the pharmaceutical composition of the present invention can be prescribed in a variety of ways depending on factors such as the formulation method, administration method, patient age, weight, sex, morbidity, diet, administration time, administration route, excretion rate, and reaction sensitivity. A skilled physician can usually easily determine the prescription and the desired effective dosage for treatment.

[0036] The pharmaceutical composition of the present invention can be administered in any convenient pharmaceutical dosage form, including parenteral dosage forms and enteral dosage forms.

[0037] Among them, the non-gastrointestinal dosage forms include injection dosage forms, respiratory tract dosage forms, cavity dosage forms, mucosal dosage forms, and skin dosage forms; the gastrointestinal dosage forms include tablets, granules, capsules, solutions, dry suspensions, powders, sustained-release agents, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0038] The injection dosage forms include but are not limited to intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections and intracavitary injections; the respiratory tract administration dosage forms include but are not limited to sprays, aerosols, and powder aerosols; the cavity administration dosage forms include but are not limited to suppositories, aerosols, effervescent tablets, drops, and pills, which can be used for, but are not limited to, the rectum, vagina, urethra, nasal cavity, and ear canal; the mucosal administration dosage forms include but are not limited to eye drops, nasal drops, eye ointments, gargles, sublingual tablets, adhesive tablets, and patches; the skin administration dosage forms include but are not limited to external solutions, lotions, liniments, ointments, plasters, pastes, and patches.

[0039] In some embodiments, the dosage form of the pharmaceutical composition is selected from an injectable dosage form.

[0040] In some embodiments, the injectable dosage form is selected from intracavitary injection.

[0041] In a specific embodiment, the intracavity injection is selected from an intraperitoneal injection.

[0042] The third aspect of the present invention provides any of the following methods:

[0043] (1) A method for promoting the proliferation and growth of hematopoietic stem cells in vitro, the method comprising administering selenocysteine or a derivative thereof;

[0044] (2) A method for enhancing the function of hematopoietic stem cells in vitro, the method comprising administering selenocysteine or a derivative thereof;

[0045] (3) A method for inhibiting apoptosis of hematopoietic stem cells in vitro, the method comprising administering selenocysteine or a derivative thereof;

[0046] (4) A method for promoting the proliferation and growth of hematopoietic stem cells after chemotherapy in vitro, the method comprising administering selenocysteine or a derivative thereof;

[0047] (5) A method for enhancing the function of hematopoietic stem cells after chemotherapy in vitro, the method comprising administering selenocysteine or a derivative thereof;

[0048] (6) A method for inhibiting apoptosis of hematopoietic stem cells after chemotherapy in vitro, the method comprising administering selenocysteine or a derivative thereof;

[0049] (7) A method for promoting the proliferation and growth of leukocytes after chemotherapy in vitro, the method comprising administering selenocysteine or a derivative thereof;

[0050] (8) A method for increasing platelet count after chemotherapy in vitro, the method comprising administering selenocysteine or a derivative thereof.

[0051] In some embodiments, the hematopoietic stem cells include long-term hematopoietic stem cells and short-term hematopoietic stem cells.

[0052] In some embodiments, the selenocysteine derivative includes a compound in which an alkyl group is attached to a selenium atom.

[0053] In the present invention, the term "alkyl" includes fully saturated branched or unbranched (e.g., straight chain or linear) hydrocarbon moieties containing 1 to 20 carbon atoms. Preferably, the alkyl group contains 1 to 7 carbon atoms, more preferably 1 to 4 carbon atoms. Representative examples of alkyl moieties include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl.

[0054] In a specific embodiment, the alkyl group is selected from methyl, and the selenocysteine derivative is selenomethylselenocysteine.

[0055] In some embodiments, the selenomethylselenocysteine includes L-selenomethylselenocysteine, D-selenomethylselenocysteine, and DL-selenomethylselenocysteine.

[0056] In a specific embodiment, the selenomethylselenocysteine is L-selenomethylselenocysteine.

[0057] A fourth aspect of the present invention provides any of the following applications:

[0058] (1) Application of selenocysteine or its derivatives in promoting the proliferation and growth of hematopoietic stem cells;

[0059] (2) Application of selenocysteine or its derivatives in enhancing the function of hematopoietic stem cells;

[0060] (3) Application of selenocysteine or its derivatives in inhibiting apoptosis of hematopoietic stem cells;

[0061] (4) Application of selenocysteine or its derivatives in promoting the proliferation and growth of hematopoietic stem cells after chemotherapy;

[0062] (5) Application of selenocysteine or its derivatives in enhancing the function of hematopoietic stem cells after chemotherapy;

[0063] (6) Application of selenocysteine or its derivatives in inhibiting apoptosis of hematopoietic stem cells after chemotherapy;

[0064] (7) Application of selenocysteine or its derivatives in promoting leukocyte proliferation and growth after chemotherapy;

[0065] (8) Application of selenocysteine or its derivatives in increasing platelet count after chemotherapy;

[0066] (9) The use of selenocysteine or its derivatives in the preparation of drugs for prolonging survival after chemotherapy.

[0067] In some embodiments, the selenocysteine derivative includes a compound in which an alkyl group is attached to a selenium atom.

[0068] In the present invention, the term "alkyl" includes fully saturated branched or unbranched (e.g., straight chain or linear) hydrocarbon moieties containing 1 to 20 carbon atoms. Preferably, the alkyl group contains 1 to 7 carbon atoms, more preferably 1 to 4 carbon atoms. Representative examples of alkyl moieties include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl.

[0069] In a specific embodiment, the alkyl group is selected from methyl, and the selenocysteine derivative is selenomethylselenocysteine.

[0070] In some embodiments, the selenomethylselenocysteine includes L-selenomethylselenocysteine, D-selenomethylselenocysteine, and DL-selenomethylselenocysteine.

[0071] In a specific embodiment, the selenomethylselenocysteine is L-selenomethylselenocysteine.

[0072] The present invention has the following advantages and beneficial effects:

[0073] The present invention provides the use of selenocysteine or its derivatives in resisting chemotherapy damage. Experiments have demonstrated that selenocysteine or its derivatives can promote the proliferation and growth of hematopoietic stem cells, enhance hematopoietic stem cell function, and inhibit hematopoietic stem cell apoptosis. Furthermore, selenocysteine can resist chemotherapy damage, promote the recovery of hematopoietic stem cells, white blood cells, and platelets after chemotherapy, and prolong survival after chemotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1 This is the experimental result of L-methylselenocysteine improving the function of mouse hematopoietic stem cells;

[0075] Figure 2 This is the experimental result of L-methylselenocysteine improving the function of hematopoietic stem cells in mice after chemotherapy;

[0076] Figure 3 This is the experimental result of L-methylselenocysteine improving the survival rate of mice after chemotherapy;

[0077] Figure 4 This is the experimental result of L-methylselenocysteine improving the recovery rate of white blood cells in mice after chemotherapy;

[0078] Figure 5 This is the experimental result of L-methylselenocysteine improving the platelet recovery rate in mice after chemotherapy;

[0079] Figure 6 This is the experimental result showing that dimethyl selenium cannot improve the recovery ability of hematopoietic stem cells in mice after chemotherapy;

[0080] Figure 7 This is the experimental result showing that dimethyl selenium cannot improve the survival rate of mice after chemotherapy;

[0081] Figure 8 This is the experimental result showing that dimethyl selenium cannot increase the recovery rate of white blood cells in mice after chemotherapy;

[0082] Figure 9 This is the experimental result showing that dimethyl selenium cannot increase the platelet recovery rate in mice after chemotherapy. DETAILED DESCRIPTION

[0083] The present invention will be further described below with reference to the following embodiments. The following description is merely a preferred embodiment of the present invention and does not limit the present invention in any other form. Any person skilled in the art may utilize the above disclosed technical content to make equivalent embodiments with equivalent variations. Any simple modification or equivalent variation of the following embodiments made in accordance with the technical essence of the present invention without departing from the content of the present invention shall fall within the scope of protection of the present invention.

[0084] Example 1 L-methylselenocysteine protects against chemotherapy damage

[0085] Experiment 1 Intraperitoneal injection of L-methylselenocysteine

[0086] Twelve wild-type C56bl / 6 mice were purchased from an animal center and randomly divided into two groups, an experimental group and a control group, with six mice in each group. The experimental group received intraperitoneal injections of 5 μg / kg of L-methylselenocysteine (MSC) once daily for 10 days, while the control group received an equal volume of solvent daily for a total of 10 injections. Ten days later, all mice were sacrificed by cervical dislocation. Tibias and femurs were dissected, and bone marrow was removed from the medullary cavity using a 1 ml syringe and dispersed into single cells using a pipette. The cell pellet was collected by centrifugation and resuspended in an appropriate amount of red blood cell lysis buffer. Lysis was continued on ice for 3 minutes. The cell pellet was collected by centrifugation, resuspended in an appropriate volume of red blood cell lysis buffer, and a cell aliquot was collected for cell counting. The remaining cells were labeled with the commonly used hematopoietic stem cell markers Lin-Sca-1+ c-Kit+ CD34-CD135- and Lin-Sca+ c-Kit+ CD34+CD135- to mark long-term hematopoietic stem cells and short-term hematopoietic stem cells, respectively. Hematopoietic stem cells within bone marrow mononuclear cells were quantified using flow cytometry. Specifically, 5 million bone marrow cells were added with 11.4 μL of a mixture of Lin Sca-1 c-Kit CD34 CD135 antibodies and incubated on ice for 1 hour. The cells were then washed once with 1 mL of PBS and centrifuged at 600 g for 5 minutes. The supernatant was discarded, and the cell pellet was resuspended in 500 μL of PBS. Another 5 million cells were added with 9.6 μL of a mixture of Lin Sca-1 c-Kit CD34 CD135 DAPI Apotracker™ Green antibodies and incubated on ice for 1 hour. The cells were then washed once with 1 mL of PBS and centrifuged at 600 g for 5 minutes. The supernatant was then discarded and the cell pellet was resuspended in 500 μl of PBS. Flow cytometry was then used for analysis. Graphpad 8.0 was used for t-test analysis. P < 0.05 was considered significant, ** P < 0.01, and *** P < 0.001.

[0087] The results are as follows Figure 1 As shown in the data, after 10 days of intraperitoneal injection of MSCs into mice, the number of long-term and short-term hematopoietic stem cells in mice increased significantly, and the apoptosis rates of long-term and short-term hematopoietic stem cells decreased significantly, indicating that supplementing with an appropriate amount of MSCs can significantly improve the function of hematopoietic stem cells in mice.

[0088] Experiment 2: Using the chemotherapy drug 5-fluorouracil to induce chemical damage to hematopoietic stem cells, it was found that after supplementation with MSCs, the recovery ability of the hematopoietic stem cells in mice was significantly enhanced.

[0089] Eight wild-type C57BL / 6 mice aged 6-8 weeks were randomly divided into two groups: a control group (n=4) and an experimental group (n=4). All eight mice were intraperitoneally injected with 150 mg / kg of 5-fluorouracil. On the second day, the experimental group received intraperitoneal injections of MSCs at a dose of 5 μg / kg once daily for 10 days. The control group received an equal volume of solvent intraperitoneally. Ten days later, all eight mice were sacrificed by cervical dislocation, and femurs and tibias were obtained as in Experiment 1. Graphpad 8.0 was used for t-test analysis. Significance was indicated as P < 0.05, **P < 0.01, and ***P < 0.001.

[0090] The results are as follows Figure 2 As shown in the data, 11 days after 5-fluorouracil treatment, the number of long-term and short-term hematopoietic stem cells in mice supplemented with MSCs was significantly higher than that in the control group, indicating that supplementation with MSCs can significantly improve the recovery ability of hematopoietic stem cells after 5-fluorouracil treatment and resist chemotherapy damage.

[0091] Experiment 3: Mice supplemented with methylselenocysteine had significantly longer survival times than mice in the control group.

[0092] Twenty 6-8 week old c57bl / 6 wild-type mice were selected and injected with 5-fluorouracil at a dose of 150 mg / kg by intraperitoneal injection. They were then randomly divided into two groups, a control group of 10 mice and an experimental group of 10 mice. On the second day, MSCs were injected intraperitoneally into the experimental group mice at a dose of 5 ug / kg once a day until the end of the experiment. The control group mice were injected with an equal amount of solvent every day. During the experimental period, all mice were injected with 5-fluorouracil once every seven days for two consecutive weeks, for a total of three injections. The survival period of the mice was recorded, and the results are as follows: Figure 3 We used GraphPad 8.0 to perform t-tests, with significance indicated by P < 0.05 and ***P < 0.001.

[0093] The results showed that after continuous injection of 5-fluorouracil, mice supplemented with MSCs had a longer survival period, indicating that supplementing with MSCs can resist 5-fluorouracil-induced chemotherapy damage and improve the survival rate of chemotherapy mice.

[0094] Experiment 4: Supplementation of MSCs can significantly increase the recovery rate of white blood cells damaged by chemotherapy drugs

[0095] Twelve 6-8 week old c57bl / 6 wild-type mice were randomly divided into two groups, a control group of 6 mice and an experimental group of 6 mice. These 12 mice were intraperitoneally injected with 5-fluorouracil at a dose of 150mg / kg. On the second day, the experimental group mice were intraperitoneally injected with MSC at a dose of 5ug / kg, while the control group was injected with an equal amount of solvent. The injections were continued once a day until the end of the experiment. On the 0th, 1st, 5th and 11th day of the experiment, venous blood was drawn from the mice into anticoagulant tubes, and the number of white blood cells in the venous blood was detected by routine blood tests. The results are as follows: Figure 4 We used GraphPad 8.0 for t-test, with significance indicated by P < 0.05 and ***P < 0.001.

[0096] The results showed that the number of white blood cells in the experimental group increased significantly compared with the control group, indicating that supplementing with methylselenocysteine can significantly increase the recovery rate of white blood cells in mice.

[0097] Experiment 5: Supplementation of methylselenocysteine can significantly restore the recovery rate of platelets damaged by chemotherapy drugs

[0098] The experimental method was the same as that of Experiment 4 in this embodiment. Venous blood was collected from mice on the 0th, 1st, 5th and 11th day of the experiment, and the platelet count was detected by routine blood tests. The results were as follows: Figure 5 We used GraphPad 8.0 to perform t-tests, with significance indicated by P < 0.05 and ***P < 0.001.

[0099] The results showed that the number of platelets in the experimental group increased significantly compared with the control group, indicating that supplementing with methylselenocysteine can significantly increase the platelet recovery rate in mice.

[0100] Example 2 Dimethylselenium cannot resist chemotherapy damage

[0101] Experiment 1 used the chemotherapy drug 5-fluorouracil to induce chemical damage to hematopoietic stem cells, and found that after supplementation with dimethylselenium (DMSe), the recovery ability of the hematopoietic stem cells of mice did not change significantly.

[0102] Eight 6-8-week-old C57BL / 6 wild-type mice were randomly divided into two groups: a control group (n=4) and an experimental group (n=4). All eight mice were intraperitoneally injected with 150 mg / kg of 5-fluorouracil. On the second day, the experimental group received an intraperitoneal injection of 5 μg / kg of DMSe once daily for 10 days. The control group received an equal volume of solvent intraperitoneally. Ten days later, all eight mice were sacrificed by cervical dislocation. Femurs and tibias were removed, and bone marrow was removed from the medullary cavity using a 1 ml syringe and dispersed into single cells using a pipette. The cell pellet was collected by centrifugation and resuspended in an appropriate amount of red blood cell lysis buffer. The cells were lysed on ice for 3 minutes. The cell pellet was collected by centrifugation, resuspended in an appropriate volume of red blood cell lysis buffer, and a portion of the cells was collected for cell counting. The remaining cells were labeled with the commonly used hematopoietic stem cell markers Lin-Sca-1+ c-Kit+ CD34-CD135- for long-term hematopoietic stem cells and Lin-Sca+ c-Kit+CD34+CD135- for short-term hematopoietic stem cells, respectively. Hematopoietic stem cells within bone marrow mononuclear cells were quantified using flow cytometry. Specifically, 5 million bone marrow cells were collected and 11.4 μl of a mixture of Lin-Sca-1, c-Kit, CD34, and CD135 antibodies was added. The cells were incubated on ice for 1 hour. The cells were then washed once with 1 ml of PBS and centrifuged at 600 g for 5 minutes. The supernatant was discarded, and the cell pellet was resuspended in 500 μl of PBS. Flow cytometric analysis was performed using t-tests in GraphPad 8.0. P < 0.05 was considered significant, and ns indicated no significant difference.

[0103] The results are as follows Figure 6 As shown in the figure, after the 11th day of 5-fluorouracil treatment, the number of long-term hematopoietic stem cells and short-term hematopoietic stem cells in the mice in the DMSe supplementation group did not change significantly compared with the control group, indicating that DMSe supplementation cannot significantly improve the recovery ability of hematopoietic stem cells after 5-fluorouracil treatment.

[0104] Experiment 2: There was no significant difference in the survival time of mice supplemented with DMSe compared with those in the control group.

[0105] Twenty wild-type C57BL / 6 mice aged 6-8 weeks were injected intraperitoneally with 150 mg / kg of 5-fluorouracil. They were then randomly divided into two groups: a control group of 10 mice and an experimental group of 10 mice. On the second day, the experimental group received an intraperitoneal injection of DMSe at a dose of 5 μg / kg once daily for the remainder of the experiment. The control group received an equal volume of solvent daily. During the experimental period, all mice received a 5-fluorouracil injection every seven days for two consecutive weeks, for a total of three injections. Mouse survival was recorded. t-tests were performed using GraphPad 8.0. P < 0.05 was considered significant, and ns indicated no significant difference.

[0106] The results are as follows Figure 7 As shown: After continuous injection of 5-fluorouracil, the survival time of mice supplemented with DMSe did not change significantly compared with that of control mice, indicating that DMSe supplementation could not resist 5-fluorouracil-induced chemotherapy damage.

[0107] Experiment 3: Supplementation of DMSe does not significantly increase the recovery rate of leukocytes damaged by chemotherapy drugs

[0108] Twelve 6-8-week-old C57BL / 6 wild-type mice were randomly divided into two groups: a control group (n=6) and an experimental group (n=6). All 12 mice received an intraperitoneal injection of 150 mg / kg of 5-fluorouracil. The next day, the experimental group received an intraperitoneal injection of 5 μg / kg of DMSe, while the control group received an equivalent amount of solvent. Subsequent injections were continued daily until the end of the experiment. Venous blood was drawn from the mice on days 0, 1, 5, and 11 of the experiment, and the number of white blood cells in the blood was determined using a routine blood count. Student's t-test was performed using GraphPad 8.0. P < 0.05 was considered significant, and ns indicates no significant difference.

[0109] The results are as follows Figure 8 As shown, the number of white blood cells in the DMSe supplemented group did not change significantly compared with the control group, indicating that DMSe supplementation cannot significantly increase the recovery rate of white blood cells in mice after chemotherapy damage.

[0110] Experiment 4: Supplementation with DMSe does not significantly restore the recovery rate of platelets damaged by chemotherapy drugs

[0111] The experimental method was the same as that of Experiment 3. Venous blood was collected from mice on day 0, day 1, day 5, and day 11 of the experiment, and the platelet count was detected by routine blood tests. We used GraphPad 8.0 for t-tests, with P < 0.05 indicating significance, and ns indicating no significant difference.

[0112] The results are as follows Figure 9As shown: There was no significant change in the number of platelets in the DMSe supplemented group compared with the control group, indicating that DMSe supplementation could not significantly increase the platelet recovery rate in mice.

[0113] The above embodiments are only provided for understanding the method and core concept of the present invention. It should be noted that, without departing from the principles of the present invention, a number of improvements and modifications may be made to the present invention by a person skilled in the art, and such improvements and modifications shall fall within the scope of protection of the claims of the present invention.

Claims

1. Application of selenocysteine or its derivatives in the preparation of drugs for resisting chemotherapy damage.

2. The use according to claim 1, characterized in that The selenocysteine derivatives include compounds formed by linking an alkyl group to a selenium atom.

3. The use according to claim 2, characterized in that The alkyl group includes methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl; Preferably, the alkyl group is selected from methyl, and the selenocysteine derivative is selenomethylselenocysteine; Preferably, the selenomethylselenocysteine is L-selenomethylselenocysteine.

4. The use according to any one of claims 1 to 3, characterized in that The selenocysteine or its derivatives include selenocysteine or its derivatives themselves, hydrates, pharmaceutically acceptable salts, solvates or crystal forms.

5. The use according to any one of claims 1 to 3, characterized in that The chemotherapy comprises administering one or more of the following chemotherapy drugs: alkylating agents, nitrosoureas, ethyleneimines, methylamines, alkyl sulfonates, antimetabolites, vinca alkaloids, epipodophyllotoxins, antibiotics, mitomycin C, actinomycin, platinum coordination complexes, anthracenediones, methylhydrazine derivatives, hormones, and antagonists; Preferably, the chemotherapy drug is selected from the group consisting of antimetabolites; Preferably, the antimetabolites include folic acid analogs, pyrimidine analogs, and purine analogs; Preferably, the folic acid analogs include methotrexate and trimetrexate; Preferably, the pyrimidine analogs include 5-fluorouracil, capecitabine, fluorodeoxyuracil, gemcitabine, and cytarabine; Preferably, the purine analogs include 6-mercaptopurine, 6-thioguanine, and azathioprine; Preferably, the chemotherapy drug is selected from 5-fluorouracil.

6. The use according to any one of claims 1 to 3, characterized in that The chemotherapy damage includes bone marrow suppression, kidney disease, liver disease, gastrointestinal disease, mucosal damage, dry mouth, neurotoxicity, cardiotoxicity, ototoxicity, alopecia, and pulmonary fibrosis; Preferably, the chemotherapy injury is selected from myelosuppression; Preferably, the bone marrow suppression includes reduction or functional impairment of hematopoietic stem cells, red blood cells, white blood cells, and thrombocytocytes; Preferably, the hematopoietic stem cells include long-term hematopoietic stem cells and short-term hematopoietic stem cells.

7. The use according to any one of claims 1 to 3, characterized in that The pharmaceutically effective amount of selenocysteine or its derivatives is 0.1-10 μg / kg; Preferably, the pharmaceutically effective amount of selenocysteine or its derivatives is 0.55 μg / kg.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition includes selenocysteine or its derivatives, which can resist chemotherapy damage; Preferably, the selenocysteine derivative includes a compound formed by connecting an alkyl group to a selenium atom; Preferably, the alkyl group includes methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl; Preferably, the alkyl group is selected from methyl, and the selenocysteine derivative is selenomethylselenocysteine; Preferably, the selenocysteine or its derivatives include selenocysteine or its derivatives themselves, hydrates, pharmaceutically acceptable salts, solvates or crystalline forms; Preferably, the pharmaceutical composition further comprises other drugs with anti-chemotherapy damage effects and organ protection drugs; Preferably, the other drugs with anti-chemotherapy damage effects and organ protection drugs include batyl alcohol, leucine, vitamin B12, folic acid, iron, hematopoietic factors, and cytokines; Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

9. Any of the following methods: (1) A method for promoting the proliferation and growth of hematopoietic stem cells in vitro, characterized in that: The method comprises administering selenocysteine or a derivative thereof; (2) A method for enhancing the function of hematopoietic stem cells in vitro, characterized in that the method comprises administering selenocysteine or a derivative thereof; (3) A method for inhibiting apoptosis of hematopoietic stem cells in vitro, characterized in that the method comprises administering selenocysteine or a derivative thereof; (4) A method for promoting the proliferation and growth of hematopoietic stem cells after chemotherapy in vitro, characterized in that the method comprises administering selenocysteine or a derivative thereof; (5) A method for enhancing the function of hematopoietic stem cells after chemotherapy in vitro, characterized in that the method comprises administering selenocysteine or a derivative thereof; (6) A method for inhibiting apoptosis of hematopoietic stem cells after chemotherapy in vitro, characterized in that the method comprises administering selenocysteine or a derivative thereof; (7) A method for promoting the proliferation and growth of leukocytes after chemotherapy in vitro, characterized in that the method comprises administering selenocysteine or a derivative thereof; (8) A method for increasing platelet count after chemotherapy in vitro, characterized in that the method comprises administering selenocysteine or a derivative thereof; Preferably, the hematopoietic stem cells include long-term hematopoietic stem cells and short-term hematopoietic stem cells; Preferably, the selenocysteine derivative includes a compound formed by connecting an alkyl group to a selenium atom; Preferably, the alkyl group includes methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl; Preferably, the alkyl group is selected from methyl, and the selenocysteine derivative is selenomethylselenocysteine; Preferably, the selenomethylselenocysteine is L-selenomethylselenocysteine.

10. Any of the following applications: (1) Application of selenocysteine or its derivatives in promoting the proliferation and growth of hematopoietic stem cells; (2) Application of selenocysteine or its derivatives in enhancing the function of hematopoietic stem cells; (3) Application of selenocysteine or its derivatives in inhibiting apoptosis of hematopoietic stem cells; (4) Application of selenocysteine or its derivatives in promoting the proliferation and growth of hematopoietic stem cells after chemotherapy; (5) Application of selenocysteine or its derivatives in enhancing the function of hematopoietic stem cells after chemotherapy; (6) Application of selenocysteine or its derivatives in inhibiting apoptosis of hematopoietic stem cells after chemotherapy; (7) Application of selenocysteine or its derivatives in promoting leukocyte proliferation and growth after chemotherapy; (8) The use of selenocysteine or its derivatives in increasing platelet count after chemotherapy; (9) Use of selenocysteine or its derivatives in the preparation of drugs for prolonging survival after chemotherapy; Preferably, the selenocysteine derivative includes a compound formed by connecting an alkyl group to a selenium atom; Preferably, the alkyl group includes methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl; Preferably, the alkyl group is selected from methyl, and the selenocysteine derivative is selenomethylselenocysteine; Preferably, the selenomethylselenocysteine is L-selenomethylselenocysteine.

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