CN112437673A - Use of sulphoxides for the prophylaxis of diseases of the radioactive nervous system - Google Patents

Abstract

The invention provides an application of sulfoxide compound in preparing medicine for preventing human suffering from head and neck tumor to be treated by radiation from suffering from radioactive nervous system disease. The sulfoxide compound has excellent effect in preventing human suffering from head and neck tumor from suffering from radioactive nervous system diseases.

Description

Use of sulphoxides for the prophylaxis of diseases of the radioactive nervous system

The present application claims priority of chinese application 201810800390.9 entitled "use of dimethyl sulfoxide in preventing radiation oral mucositis" filed on 20/7/2018, the contents of which are incorporated herein by reference.

Technical Field

The invention relates to the field of medicine, in particular to an application of sulfoxide compounds, especially dimethyl sulfoxide in preventing occurrence of radioactive nervous system diseases in a tumor subject to be treated by radiation.

Background

Radiotherapy is one of the common treatment methods for head and neck malignant tumors, and can bring great benefits to patients regardless of primary brain tumors or brain metastases, and is the preferred treatment method for radiotherapy for patients with nasopharyngeal carcinoma. The nervous system is one of the major dose-limiting tissues and organs for tumor radiotherapy. Radiation-induced nerve System Disease (Radiation-induced nerve System Disease) refers to an adverse reaction of tissues generated by ionizing Radiation of each part of the Nervous System, is one of the main complications of radiotherapy such as head and neck malignant tumors, cerebrovascular malformations, thoracic and abdominal tumors and is occasionally caused by ionizing Radiation accidents. The radioactive nervous system diseases are generally divided into an acute stage, an early-late stage and a late-late stage, namely the acute stage usually occurs days to 1 month after irradiation; the early-late response period usually occurs 1 to 6 months after irradiation; late delayed response periods often occur 6 months after irradiation. Clinically, the symptoms of lethargy, anorexia, apathy, headache, memory decline, vertigo, nausea, emesis, one-side limb muscle decline, ataxia aggravation, enuresis, etc. are presented.

The radioactive nervous system disease is one of the most serious complications after tumor radiotherapy, the pathogenesis is not completely clear, multiple mechanisms such as neuron and glial cell injury, angiopathy, immune inflammatory reaction and the like can be involved, and specific prevention and treatment medicines are not available. Traditional treatment methods such as hyperbaric oxygen, glucocorticoid, surgery and the like can effectively relieve clinical symptoms of patients, but can not reverse the damage process of the nervous system, and most patients still suffer from cognitive, directional, intelligent and memory disorders after meeting, and therefore lose labor capacity and social interaction capacity.

In view of these drawbacks, there is still a need to develop new drugs that are effective in preventing the occurrence of radioactive nervous system diseases in tumor subjects to be treated with radiation.

Disclosure of Invention

In one aspect, the invention provides the use of a sulphoxide compound in the manufacture of a medicament for the prevention of a radiologic neurological disease in a subject suffering from a tumour to be treated with radiation.

In another aspect, the present invention provides a sulfoxide compound or a pharmaceutical composition comprising a sulfoxide compound for use in preventing a radioactive nervous system disorder in a subject to be treated with radiation.

In a further aspect, the present invention provides a method of preventing a radioactive nervous system disease using a sulfoxide compound or a pharmaceutical composition comprising the sulfoxide compound, comprising the steps of: administering a sulfoxide compound or a pharmaceutical composition comprising a sulfoxide compound to a subject having a tumor to be treated with radiation.

In one embodiment, the sulfoxides are DMSO or tetramethylene sulfoxide, preferably DMSO.

In one embodiment, the radioactive nervous system disorder comprises radioactive brain injury, radioactive spinal cord injury, and radioactive peripheral nerve injury.

In one embodiment, the radioactive brain injury is selected from at least one of: acute radioactive brain injury, early late radioactive brain injury, and late radioactive brain injury.

In one embodiment, the radioactive brain injury manifests as one or more symptoms selected from the group consisting of: cerebral edema, lethargy syndrome, and mental retardation.

In one embodiment, the radioactive spinal cord injury is selected from at least one of: chronic progressive radiation spinal cord injury, muscle-collapsing radiation spinal cord injury, acute radiation spinal cord injury, and transient radiation spinal cord injury.

In one embodiment, the radioactive peripheral nerve injury is selected from at least one of: nerve cell injury, neural stem cell injury, neuronal precursor cell injury, hippocampal neural stem cell injury, hippocampal neuronal precursor cell injury, neurite injury, nerve fiber injury and neurosynaptic injury.

In one embodiment, the radiologic neurological disease manifests as one or more symptoms selected from the group consisting of: memory impairment, memory deficit, cognitive disorder, cognitive impairment, and learning impairment.

In one embodiment, the subject having a tumor is a subject having one or more tumors selected from the group consisting of: tumors of the digestive system, such as oral cancer, tonsil cancer, esophageal cancer, rectal cancer, colon cancer, liver cancer, pancreatic cancer, gastric cancer, etc.; tumors of the respiratory system, such as nasopharyngeal carcinoma, cancer of maxillary sinus, laryngeal carcinoma, bronchial cancer, lung cancer, etc.; genitourinary system tumor such as renal clear cell carcinoma, bladder cancer, nephroblastoma, testicular tumor, cervical cancer, uterine corpus cancer, etc.; breast cancer; tumors of the nervous system, such as brain tumor, medulloblastoma, neuroblastoma, pituitary tumor, etc.; malignant tumors of skin and soft tissue, such as penile cancer, lip cancer, melanoma, sarcoma; malignant tumor of bone, such as osteosarcoma, Ewing's tumor, etc.; and lymphoid neoplasms.

In one embodiment, the subject having a tumor is a subject having a tumor of the nervous system, preferably a human having a tumor of the nervous system.

In one embodiment, the head and neck tumor is selected from one or more of the following: brain tumors, medulloblastoma, neuroblastoma, and pituitary tumors.

In one embodiment, the subject having a tumor is a subject having a head and neck tumor, preferably a human having a head and neck tumor.

In one embodiment, the head and neck tumor comprises one or more selected from the group consisting of: neck tumors, otorhinolaryngological tumors, and oral maxillofacial tumors. In one embodiment, the neck tumor is a thyroid tumor. In one embodiment, the otorhinolaryngological tumor is a nasopharyngeal carcinoma, a laryngeal carcinoma, or a paranasal sinus carcinoma. In one embodiment, the oromaxillofacial tumor is an oral cancer, such as a tongue cancer, a gum cancer, a buccal cancer, a maxillary sinus cancer, and a tonsil cancer.

In one embodiment, the subject having a tumor is a head and neck tumor subject that will receive radiation therapy within 8 hours, preferably within 4 hours, more preferably within 2 hours, and most preferably within 1 hour.

In one embodiment, the subject having a tumor is a head and neck tumor subject that will receive chemotherapy within 8 hours, preferably within 4 hours, more preferably within 2 hours, and most preferably within 1 hour.

In one embodiment, the medicament is in a dosage form suitable for administration selected from one of: intravenous administration, oromucosal administration, transdermal administration and oral administration.

In one embodiment, the drug is in a dosage form selected from the group consisting of: infusion solution, injection, collutory, pellicle, aerosol, tablet, adhesive, patch, sublingual agent, disintegrating agent, stick, powder, ointment, gel, water solution, suspension and capsule.

In one embodiment, the medicament is administered in a dose of 0.1 to 10.0g of the sulfoxides, preferably 0.2 to 5.0g of sulfoxides, more preferably 0.4 to 2.0g of sulfoxides, most preferably 1.0g of sulfoxides per kg of body weight.

In one embodiment, the medicament further comprises one or more compounds selected from the group consisting of: WR2721, PrC-210(aminothiol), WR-1065, WR-3689, WR-151327, WR-638, WR-77913, WR-44923, cysteamine, AET, lipoic acid, N-diethyllipoamide, Mn-SOD transgenic preparations, Cu.Zn-SOD preparations, Fe-SOD preparations, GC4419(M40403), MnBuOE, AEOL10150, MnTHex-2-PyP 5+ (hexyl), MnTE-2-PyP, EUK-207, EUK-189, EUK-134, melatonin, glutathione, pentoxifylline, Edaravone (Edaravone), Tempol (4-hydro-2, 2,6,6-tetramethylpiperidine-N-oxyl), glucocorticoids such as betamethasone, betamethasone (Prednisone), anticoagulants such as acetate, anticoagulants such as heparin, and diuretics such as mannitol, and diuretics, such as ganglioside, oxiracetam, cytidine disodium triphosphate, etc., vitamins, such as B vitamins, vitamin E, blood vessel expanding medicines, such as compound radix Salviae Miltiorrhizae and BUYANGHUANWUTANG, etc., and blood vessel endothelin long-chain factor antibodies, such as bevacizumab, hyperbaric oxygen, etc.

In one embodiment, the radiation therapy comprises a treatment with radiation selected from the group consisting of: alpha rays, beta rays, gamma rays, x rays, neutrons, electron rays, proton beams, particle beams, and combinations thereof.

In one embodiment, the radiation therapy comprises external radiation therapy and internal radiation therapy.

In one embodiment, the radiation therapy method includes conventional fractionated radiation therapy and a single bolus radiation therapy.

In one embodiment, the radiation therapy has a radiation dose of 1-50Gy, more preferably 1-8Gy, most preferably 2 Gy.

In one embodiment, the radiation therapy comprises the use of a radiosensitizer.

In one embodiment, the radiosensitizers are hypoxic cell radiosensitizers and non-hypoxic cell radiosensitizers,

wherein the hypoxic cell radiosensitizer is selected from one or more of: nitroimidazoles (Nitroidamidazoles), such as metronidazole, imadazole, MISO (Ro-07-0582), RSU-1096, Tirapazamine (Tirapazamine), and the like; and bioreductive agents such as sodium glycinediazole, nitrogen oxides, quinones and niacinamide and derivatives thereof;

wherein the non-hypoxic cell radiosensitizer is selected from one or more of: platinum drugs such as cisplatin, carboplatin, oxaliplatin, and the like; antimetabolites such as 5-FU, capecitabine, gemcitabine, and the like; topoisomerase inhibitors such as camptothecin (camptothecin), Irinotecan (Irinotecan), topotecan (topotecan), CPT417, BMS-286309, etoposide (VP-16), and the like; microtubule-stabilizing drugs such as paclitaxel, docetaxel, paclitaxel (patupilone), PBOX-15, etc.; a second mitochondria-derived caspase activator (SMAC) analog, such as Tat-SMACN7, SM-164, LCL161, etc.; tumor-targeted therapeutic drugs, such as Sorafenib (Sorafenib), Erlotinib (Erlotinib), Cetuximab (Cetuximab), Bevacizumab (Bevacizumab), trastuzumab (trastuzumab), and the like; photosensitizers such as 5-aminolevulinic acid, hematoporphyrin monomethyl ether, hematoporphyrin derivative (HPD), dimethylporphyrin ether (DHE), Photofrin II, and the like; DNA alkylating agents such as temozolomide and the like; and Chinese medicinal extracts such as irisquinone.

Drawings

FIG. 1: the protective effect of DMSO on the proliferation of 5Gy head-irradiated mouse hippocampal neural stem cells;

FIG. 2: the DMSO has the protection effect on the proliferation of 10Gy head-irradiated mouse hippocampal neural stem cells; and

FIGS. 3 a-d: DMSO has no radiation protection effect on the head and neck tumor of the tumor-bearing mice.

Detailed Description

When the inventor screens a novel radioprotectant, the inventor unexpectedly finds that the intraperitoneal administration or the oral administration of dimethyl sulfoxide (DMSO) can effectively prevent the radioactive nervous system diseases. The invention further proves that the DMSO pre-irradiation administration can effectively prevent radioactive brain injury and radioactive spinal cord injury.

In one aspect, the invention provides the use of a sulphoxide compound in the manufacture of a medicament for the prevention of a radiologic neurological disease in a subject suffering from a tumour to be treated with radiation.

In another aspect, the present invention provides a sulfoxide compound or a pharmaceutical composition comprising a sulfoxide compound for use in preventing a radioactive nervous system disorder in a subject to be treated with radiation.

In a further aspect, the present invention provides a method of preventing a radioactive nervous system disease using a sulfoxide compound or a pharmaceutical composition comprising the sulfoxide compound, comprising the steps of: administering a sulfoxide compound or a pharmaceutical composition comprising a sulfoxide compound to a subject having a tumor to be treated with radiation.

The terms mentioned herein preferably have the following meanings:

the term "sulfoxides" is a class of organic compounds comprising a "sulfoxide group" (> S ═ O) functional group, of the general formula R-S ═ O-R ', where R and R' are organic groups, such as optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted butyl, optionally substituted phenyl, optionally substituted allyl, and the like.

The term "dimethylsulfoxide" (DMSO) refers to a compound of the structure

DMSO is a common solvent, is a colorless and odorless transparent liquid at normal temperature, and has good thermal stability and low toxicity.

In one embodiment, the sulfoxides are DMSO or tetramethylene sulfoxide, preferably DMSO.

The term "subject" refers to mammals, including humans and non-humans, including, but not limited to, humans, pigs, dogs, rabbits, monkeys, cats, and the like. The subject of the invention is selected from subjects suffering from a tumor. In one embodiment, the subject is a human. Herein, the terms "subject", "patient" and "subject" are interchangeable unless specifically stated otherwise.

The term "Nervous system" refers to the Nervous tissue in the body that is responsible for the regulation of physiological functional activities and is divided into the central Nervous system and the peripheral Nervous system. Because of the anatomical location of the nervous system in the human body, radiation therapy of tumors can cause damage to the nervous system.

The term "radioactive nervous system disease" refers to an adverse reaction of tissues generated by ionizing radiation of various parts of the nervous system, and is one of the main complications of radiotherapy such as head and neck malignant tumors, cerebrovascular malformations, thoracic tumors, abdominal tumors, and the like. The radioactive nervous system diseases are classified into radioactive brain injury, radioactive spinal cord injury and radioactive peripheral nerve injury according to the difference of the injured parts.

The term "Radiation-induced Brain Injury" or "Radiation Encephalopathy" (Radiation Encephalopathy) refers to a nervous system Injury disease caused by radiotherapy, particularly radiotherapy to head and neck tumors. Radioactive brain injury is classified into acute injury, early late-onset injury and late-onset injury according to the time at which the radiotherapy response occurs.

The term "acute radiation brain injury" refers to symptoms of cerebral edema that occur during or within 1 month of the end of radiation therapy and may be manifested as cerebral edema, headache, nausea, vomiting, diarrhea, epilepsy, disturbance of consciousness, increased body temperature, and the like.

The term "early late onset radioactive brain injury" refers to symptoms that occur within weeks to 3 months after irradiation, primarily the somnolence syndrome after brain irradiation, manifested as lethargy, anorexia, low fever, apathy, headache, nausea, vomiting, dizziness, but with less common neuroparalytic symptoms.

The term "late-stage delayed radioactive brain injury" refers to irreversible, progressive, life-threatening brain injury that occurs 6 months to 2 years after irradiation, mainly resulting in neurological dysfunction. The injured part can be focal or diffuse white matter necrosis, and hypointelligence is the functional manifestation of radiation encephalopathy.

In one embodiment, the radioactive brain injury is selected from at least one of: acute radioactive brain injury, early late radioactive brain injury, and late radioactive brain injury.

In one embodiment, the radioactive brain injury manifests as one or more symptoms selected from the group consisting of: cerebral edema, lethargy syndrome, and mental retardation.

The term "Radioactive spinal cord injury", also known as Radiation myelopathy (Radiation myelopathy) or Radiation myelitis (Radiation myelitis), is an injury caused by ionizing Radiation (х, gamma, neutron, and electron beam Radiation, etc.) exceeding the spinal cord bearing range. The clinical manifestations of radioactive spinal cord injury are mainly classified into the following: chronic progressive radiation spinal cord injury, muscle-collapsing radiation spinal cord injury, acute radiation spinal cord injury, and transient radiation spinal cord injury.

The term "Radiation-induced peripheral nerve injure" refers to a radioactive nervous system disease caused by direct and indirect damage of peripheral nervous tissue by Radiation during radiotherapy. Breast cancer, cervical tumors, testicular tumors, and lymphoma radiation therapy are most likely to cause post-radiation brachial plexus and lumbosacral plexus damage; the radiotherapy of nasopharyngeal tumors can cause glossopharyngeal nerve damage; and optic nerve and optic cross injury can occur after radiotherapy of pituitary tumors and craniopharyngiomas.

In one embodiment, the radioactive nervous system disorder comprises radioactive brain injury, radioactive spinal cord injury, and radioactive peripheral nerve injury.

In one embodiment, the radioactive spinal cord injury is selected from at least one of: chronic progressive radiation spinal cord injury, muscle-collapsing radiation spinal cord injury, acute radiation spinal cord injury, and transient radiation spinal cord injury.

In one embodiment, the radiologic neurological disease manifests as one or more symptoms selected from the group consisting of: memory impairment, memory deficit, cognitive disorder, cognitive impairment, and learning impairment.

In one embodiment, the radiologic neurological disease is manifested by one or more lesions selected from the group consisting of: nerve cell injury, neural stem cell injury, neuronal precursor cell injury, hippocampal neural stem cell injury, hippocampal neuronal precursor cell injury, neurite injury, nerve fiber injury and neurosynaptic injury.

The term "tumor" refers to a neoplasm formed by local tissue cell proliferation of the body under the action of various tumorigenic factors. The definition and classification of such tumors can be found, for example, in the histological classification of tumors as established by the World Health Organization (WHO), second edition (1989-2000).

In one embodiment, the subject having a tumor is a subject having one or more tumors selected from the group consisting of: tumors of the digestive system, such as oral cancer, tonsil cancer, esophageal cancer, rectal cancer, colon cancer, liver cancer, pancreatic cancer, gastric cancer, etc.; tumors of the respiratory system, such as nasopharyngeal carcinoma, cancer of maxillary sinus, laryngeal carcinoma, bronchial cancer, lung cancer, etc.; genitourinary system tumor such as renal clear cell carcinoma, bladder cancer, nephroblastoma, testicular tumor, cervical cancer, uterine corpus cancer, etc.; breast cancer; tumors of the nervous system, such as brain tumor, medulloblastoma, neuroblastoma, pituitary tumor, etc.; malignant tumors of skin and soft tissue, such as penile cancer, lip cancer, melanoma, sarcoma; malignant tumor of bone, such as osteosarcoma, Ewing's tumor, etc.; and lymphoid neoplasms.

In one embodiment, the subject having a tumor is a subject having a tumor of the nervous system, preferably a human having a tumor of the nervous system.

The term "head and neck tumor" includes three major parts of neck tumor, otorhinolaryngological tumor and oral maxillofacial tumor according to different parts. Cervical tumors include, for example, thyroid tumors; otolaryngological tumors include, for example, laryngeal cancer, paranasal sinus cancer, and the like; oral and maxillofacial tumors include, for example, oral cancers such as tongue cancer, gum cancer, buccal cancer, and the like.

In one embodiment, the subject having a tumor is a subject having a head and neck tumor, preferably a human having a head and neck tumor. In one embodiment, the head and neck tumor comprises one or more selected from the group consisting of: neck tumors, otorhinolaryngological tumors, and oral maxillofacial tumors. In one embodiment, the neck tumor is a thyroid tumor. In one embodiment, the otorhinolaryngological tumor is a nasopharyngeal carcinoma, a laryngeal carcinoma, or a paranasal sinus carcinoma. In one embodiment, the oromaxillofacial tumor is an oral cancer, such as a tongue cancer, a gum cancer, a buccal cancer, a maxillary sinus cancer, and a tonsil cancer.

In one embodiment, the head and neck tumor is selected from one or more of the following: brain tumors, medulloblastoma, neuroblastoma, and pituitary tumors.

The term "preventing" includes reducing the risk of having, contracting, or experiencing a disease, disorder, condition, or sign, its development and/or progression, and/or its symptoms. The prophylactic effect may include partial or complete prevention of the underlying disease. In the present invention, the term prevention is not intended to mean that the disease is completely avoided. In the present invention, the term "prevention" does not include the treatment of diseases. In the present invention, the sulphoxide compound (preferably DMSO) and the drug comprising it need to be administered before the onset of the disease, in particular before the subject receives radiation therapy. In one embodiment, the subject receiving a sulfoxide compound (preferably DMSO) prior to receiving radiation therapy optionally includes the subject receiving radiation therapy and a sulfoxide compound (preferably DMSO) simultaneously. The invention proves that by administering a sulfoxide compound (preferably DMSO) to a tumor-bearing subject before radiotherapy, the tumor-bearing subject can be effectively prevented from suffering from a radioactive nervous system disease after receiving the radiotherapy.

In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 8 hours to 1 hour. In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 6 hours. In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 5 hours. In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 4 hours. In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 3 hours. In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 2 hours. In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 1 hour. In one embodiment, the tumor-bearing subject is a tumor subject that will receive radiation therapy within 30 minutes.

The term "dose" refers to the amount of sulfoxide compound (preferably DMSO) or sulfoxide compound (preferably DMSO) contained in a drug/pharmaceutical composition administered to a subject. The dosage may vary according to factors known in the art, such as the disease state, age, sex and weight of the person to be prevented.

In one embodiment, the dosage of the sulfoxides (preferably DMSO) is determined based on the body weight of the subject. In one embodiment, the sulfoxides (preferably DMSO) are administered at a dose of 0.1-10.0g/kg body weight. In one embodiment, DMSO is administered at a dose of 0.2-5.0g/kg body weight. In one embodiment, the sulphoxide compound (preferably DMSO) is administered in a dose of 0.4-2.0g/kg body weight. In one embodiment, the sulphoxide compound (preferably DMSO) is administered in a dose of 0.5g/kg body weight. In one embodiment, the sulphoxide compound (preferably DMSO) is administered in a dose of 1.0g/kg body weight. The invention proves that the sulfoxide compound (preferably DMSO) can effectively prevent the radioactive nervous system diseases, and the DMSO has low potential toxic and side effects in the effective dose range of preventive administration.

The term "radiotherapy" includes a method of treating tumor by irradiating tumor tissue with radiation, and generating free radicals by direct action on cellular DNA or interaction with intracellular water molecules, thereby causing death of tumor cells. Radiation administered by radiotherapy includes alpha, beta, gamma rays produced by radioisotopes, and x-rays, electron beams, proton beams and other particle beams produced by various types of x-ray treatment machines or accelerators, and the like. The mechanism of action of radiation therapy is different from that of chemotherapy. Herein, the terms "radiation therapy," "radiotherapy" and "radiotherapy" are interchangeable unless specifically stated otherwise. In one embodiment, the radiation therapy comprises a treatment with radiation selected from the group consisting of: alpha rays, beta rays, gamma rays, x rays, neutrons, electron rays, proton beams, particle beams, and combinations thereof.

Radiation therapy techniques are divided into external irradiation and internal irradiation. External radiation radiotherapy refers to the way that radiation enters the body from the outside through various tissues of the body, generates a radiation effect on a tumor target area, and kills tumors. Conventional external beam radiotherapy is fractionated radiotherapy (2 Gy each 5 times a week) established based on clinical practice experience and is considered as a standard method of tumor radiotherapy. Internal-irradiation radiation therapy, also known as brachytherapy, includes radioactive-particle-implant radiotherapy and radionuclide-targeted tumor therapy.

The radiation therapy subjects can be classified into a radiation-highly sensitive tumor, a radiation-moderately sensitive tumor, and a radiation-lowly sensitive tumor according to the irradiation dose. In one embodiment, the radiation therapy has an irradiation dose of 1 to 50 Gy. In one embodiment, the radiation therapy is delivered at a dose of 1-8 Gy. In one embodiment, the radiation therapy is delivered at a dose of 2 Gy.

In the present invention, the sulfoxides (preferably DMSO) may be further administered in combination with other compounds known to be prophylactically and/or therapeutically effective or potentially effective against radioactive neurological diseases. Such compounds include one or more compounds selected from the group consisting of: WR2721, PrC-210(aminothiol), WR-1065, WR-3689, WR-151327, WR-638, WR-77913, WR-44923, cysteamine, AET, lipoic acid, N-diethyllipoamide, Mn-SOD transgenic preparations, Cu.Zn-SOD preparations, Fe-SOD preparations, GC4419(M40403), MnBuOE, AEOL10150, MnTHex-2-PyP 5+ (hexyl), MnTE-2-PyP, EUK-207, EUK-189, EUK-134, melatonin, glutathione, pentoxifylline, Edaravone (Edaravone), Tempol (4-hydro-2, 2,6,6-tetramethylpiperidine-N-oxyl), glucocorticoids such as betamethasone, betamethasone (Prednisone), anticoagulants such as acetate, anticoagulants such as heparin, and diuretics such as mannitol, and diuretics, such as ganglioside, oxiracetam, cytidine disodium triphosphate, etc., vitamins, such as B vitamins, vitamin E, blood vessel expanding medicines, such as compound radix Salviae Miltiorrhizae and BUYANGHUANWUTANG, etc., and blood vessel endothelin long-chain factor antibodies, such as bevacizumab, hyperbaric oxygen, etc.

In the present invention, the sulfoxides (preferably DMSO) can be prepared into pharmaceutical dosage forms suitable for different modes of administration. In one embodiment, the medicament is in a dosage form selected from one or more modes of administration suitable for: intravenous administration, oromucosal administration, transdermal administration and oral administration.

The term "intravenous injection form", also known as infusion solution, refers to a large dose of injection solution infused into the human body by intravenous drip.

In one embodiment, the medicament is administered by intravenous injection. In one embodiment, the concentration of the sulfoxides (preferably DMSO) is administered intravenously in a range of 1% to 100%, preferably 5% to 50%, more preferably 10% to 20%, and most preferably 10%. In one embodiment, the intravenous injection formulation comprising a sulfoxide compound (preferably DMSO) may further comprise an injection diluent selected from sodium bicarbonate injection, sodium chloride injection, dextrose injection, sodium citrate injection, sodium glycerophosphate injection, preferably 1.4% sodium bicarbonate injection. In one embodiment, the intravenous formulation comprising a sulphoxide compound (preferably DMSO) may further comprise an injection adjunct, which may be selected from the group consisting of an osmotic pressure regulator (glucose, sodium chloride, phosphate or citrate, etc.); pH adjusters (hydrochloric acid, sulfuric acid, citric acid, sodium (potassium) hydroxide, sodium hydrogen carbonate, disodium hydrogen phosphate, sodium dihydrogen phosphate, etc.); suspending agent (propylene glycol, glycerol, tween-80, span-85, pluronic F-68, carboxymethyl cellulose, sodium alginate, polyvinylpyrrolidone, gelatin, mannitol, sorbitol, aluminum monostearate, silicone oil, etc.); and analgesics (benzyl alcohol (about 1%), procaine hydrochloride (0.5% -2.0%), lidocaine hydrochloride (0.2% -1.0%), chlorobutanol (0.3% -0.5%).

The term "oromucosal dosage form" refers to a dosage form in which the drug is absorbed through the oral mucosa and then enters the circulatory system directly. The medicine can be administered via oral mucosa to exert local and systemic therapeutic effects.

In the present invention, dosage forms suitable for oromucosal administration include topical therapeutic dosage forms: mouthwashes, films, aerosols and tablets, preferably mouthwashes and films; systemic therapeutic dosage forms: adhesive tablets, plasters, sublingual tablets and orally disintegrating tablets, preferably adhesive tablets and plasters.

In one embodiment, the dosage form suitable for oromucosal administration is selected from the following: mouthwash, film, aerosol, tablet, adhesive tablet, patch, sublingual tablet and orally disintegrating tablet.

In one embodiment, the medicament is in one or more dosage forms selected from the group consisting of: mouth washes (Mouthwash, or Oral rise), Films, aerosols, tablets, adhesive tablets, patches, sublingual tablets, orally disintegrating tablets, sticks, powders, ointments, suspensions, capsules, gels, Transmucosal patches (Transmucosal patches), Transmucosal colloids (Transmucosal gels), Films (Films), sprays, gels (Colloid), emulsions (Emulsion), ointments (increment), Rubber ointments (Rubber Paste), sponges (Sponge), creams (Cream), creams (Emulsion), pastes (Paste), paints and foams (Foam).

In one embodiment, the concentration of the sulfoxides (preferably DMSO) administered oromucosally ranges from 1% to 100%, preferably from 5% to 90%, more preferably from 10% to 70%, and most preferably from 10% to 50%.

The term "Transdermal Drug Delivery (TDD) is an administration mode in which a drug is continuously delivered to the skin surface or into the circulatory system by the skin as an administration route, thereby exerting the effect of local or systemic treatment of the skin.

The term "Transdermal drug delivery system" TDDS refers to a dosage form that is administered to the skin surface such that the drug passes through the layers of the skin at a constant rate (or near constant rate) and enters the systemic circulation to produce a systemic or local therapeutic effect.

The dosage form of transdermal drug delivery system includes, but is not limited to, plaster, liniment, film coating agent, cataplasm, matrix type patch, membrane-controlled patch, microemulsion, liposome, clathrate, prodrug and low-melting substance, etc., and these drugs can be applied to the thin part of skin, such as behind the ear, inside of arm, chest area, scrotum, etc.

In one embodiment, a dosage form suitable for transdermal administration may be selected from one or more of the following: plaster, liniment, plastics, cataplasma, matrix type patch, membrane controlled type patch, microemulsion, liposome and clathrate.

In one embodiment, a dosage form suitable for transdermal administration may be applied to a site of relatively thin skin, such as behind the ear, inside the arms, in the precordial region, in the scrotum, and the like.

Transdermal drug delivery systems are preferably passive transdermal drug delivery systems based on nanocarriers. In one embodiment, the nanocarrier-based passive transdermal delivery system may be selected from one or more of the following: nanovesicles (Vesicles), Lipid nanoparticles (Lipid nanoparticles), microemulsions (Microemulsion), and Polymeric nanoparticles (Polymeric nanoparticles).

Methods for promoting transdermal penetration of drugs include chemical methods and physical methods. The chemical method is mainly transdermal absorption enhancer, and the physical method includes ultrasonic method, ion introduction method, electroporation method, etc.

The penetration enhancer is a substance capable of helping the drug to diffuse through the stratum corneum and epidermis of the skin, or is called a transdermal enhancer, an absorption enhancer and the like.

In one embodiment, the medicament comprising a sulphoxide compound (preferably DMSO) may further comprise a penetration enhancer. Penetration enhancers can be classified into lipophilic lysozymes, surfactants, two-component and multi-component systems. In one embodiment, the penetration enhancer is selected from one or more of the following: laurocapram (Azone), linoleic acid, urea, alpha-pyrrolidone, propylene glycol, butylene glycol, dimethylformamide, Oleic Acid (OA), dodecanol (LA), etc.; the Chinese medicinal repairing promoter comprises oleum Menthae Dementholatum, oleum Eucalypti, Borneolum Syntheticum, oleum Terebinthinae, menthol, Camphora, Mentholum, etc.; dodecyl N, N-dimethylaminoisopropionate (DDAIP) and dodecyl N, N-dimethylaminoacetate (DDAA).

In one embodiment, the concentration of the sulfoxides (preferably DMSO) administered transdermally is in the range of 1% to 100%, preferably 5% to 90%, more preferably 25% to 70%, and most preferably 60%.

The term "oral dosage form" includes aqueous solutions, aqueous suspensions, tablets and capsules. In one embodiment, the dosage form for oral administration comprising a sulfoxide compound (preferably DMSO) may further comprise additional agents such as flavoring agents, bacteriostats, and the like. The term "flavoring agent" refers to pharmaceutical excipients used in pharmaceuticals to improve or mask the unpleasant taste and odor of drugs, making it difficult for patients to detect the strong bitter taste (or other off-flavors such as pungent, etc.) of drugs, and generally includes four classes of sweeteners, aromatics, mucilages, and effervescent agents.

In one embodiment, the concentration of the sulfoxides (preferably DMSO) is administered orally in a range of 1% to 100%, preferably 10% to 90%, more preferably 20% to 70%, and most preferably 50%.

In one embodiment, the medicament is administered in a dose of 0.1-10.0g DMSO/kg body weight, preferably 0.2-5.0g sulfoxides (preferably DMSO)/kg body weight, more preferably 0.4-2.0g sulfoxides (preferably DMSO)/kg body weight, most preferably 0.5-1.0g sulfoxides (preferably DMSO)/kg body weight.

In the present invention, the drug comprising a sulfoxide compound (preferably DMSO) may further comprise a pharmaceutically acceptable adjuvant. In one embodiment, the pharmaceutically acceptable excipient is selected from one or more of the following: d-panthenol (Dexpanthenol), sodium alginate, Povidone iodine (Povidone iodine), sodium hyaluronate, pectin, viscous xylan, and Sucralfate (Sucralfate). Herein, the terms "drug" and "pharmaceutical composition" are interchangeable unless specifically stated otherwise.

In one embodiment, the drug comprising a sulfoxide compound (preferably DMSO) may further be administered simultaneously with the radiosensitizer. The term "radiosensitizer" is a chemical or pharmaceutical agent that, when applied concurrently with radiation therapy, alters the reactivity of tumor cells to radiation, thereby increasing the killing effect on tumor cells, and includes hypoxic cell radiosensitizers and non-hypoxic cell radiosensitizers.

In one embodiment, the radiosensitizer is a hypoxic cell radiosensitizer or a non-hypoxic cell radiosensitizer. In one embodiment, the hypoxic cell radiosensitizer is selected from the group consisting of: nitroimidazoles (Nitroidamidazoles), such as metronidazole, imadazole, MISO (Ro-07-0582), RSU-1096, Tirapazamine (Tirapazamine), and the like; and bioreductive agents such as sodium glycin diazole, nitrogen oxides, quinones and niacinamide and derivatives thereof. In one embodiment, the non-hypoxic cell radiosensitizer is selected from the group consisting of: platinum drugs such as cisplatin, carboplatin, oxaliplatin, and the like; antimetabolites such as 5-FU, capecitabine, gemcitabine, and the like; topoisomerase inhibitors such as camptothecin (camptothecin), Irinotecan (Irinotecan), topotecan (topotecan), CPT417, BMS-286309, etoposide (VP-16), and the like; microtubule-stabilizing drugs such as paclitaxel, docetaxel, paclitaxel (patupilone), PBOX-15, etc.; a second mitochondria-derived caspase activator (SMAC) analog, such as Tat-SMACN7, SM-164, LCL161, etc.; tumor-targeted therapeutic drugs, such as Sorafenib (Sorafenib), Erlotinib (Erlotinib), Cetuximab (Cetuximab), Bevacizumab (Bevacizumab), trastuzumab (trastuzumab), and the like; photosensitizers such as 5-aminolevulinic acid, hematoporphyrin monomethyl ether, hematoporphyrin derivative (HPD), dimethylporphyrin ether (DHE), Photofrin II, and the like; DNA alkylating agents such as temozolomide and the like; and Chinese medicinal extracts such as irisquinone.

The invention also provides the use of a sulphoxide compound, preferably DMSO, in the manufacture of a medicament for the prevention of a radiologic neurological disease in a subject suffering from a tumour to be treated with a combination of radiotherapy and chemotherapy.

The term "chemotherapy treatment" is also known as "chemotherapy" or "chemotherapy," which refers to the use of chemotherapeutic drugs to kill cancer cells for therapeutic purposes. The term "combination radiation and chemotherapy" refers to a method of treating tumors using both radiation therapy and chemotherapy. Radiotherapy and chemotherapy combined therapy is one of the most widely applied treatment means in clinical tumor treatment. Radiotherapy and chemotherapy are combined to directly attack the focus of tumor, and the chemotherapy destroys cancer cells in the body of the patient, so as to achieve the treatment effect. In the present invention, it is contemplated that a subject to be treated will receive both radiation therapy and chemotherapy, either radiation therapy followed by chemotherapy, or chemotherapy followed by radiation therapy; provided that the above treatments are all performed before receiving the sulphoxide compound (preferably DMSO) of the invention.

In one embodiment, the subject with the tumor will receive treatment with one or more chemotherapeutic drugs selected from the group consisting of: alkylating agents or platinum compounds acting directly on DNA, such as melphalan (melphalan), cyclophosphamide (cyclophosphamide), oxazaphosphamide (oxazaphosphorine), cisplatin (cispin), carboplatin (carboplatin), oxaliplatin (oxaliplatin), satraplatin (satraplatin), tetraplatin (tetraplatin), iproplatin (iproplatin), mitomycin (mitomycin), streptozocin (streptozocin), carmustine (carmustine) (BCNU), lomustine (ccu), busulfan (busufan), ifosfamide (ifosfamide), streptozocin (streptozocin), thiotepa (thiotepa), chlorambucil (chloreucil); nitrogen mustards, such as dichloromethyl diethylamine (mechlorothiamine); an ethyleneimine compound; alkyl sulfonates (alkylsulfonates); antimetabolites that interfere with DNA and RNA synthesis, such as cytarabine (cytarabine), 5-fluorouracil (5-FU), pemetrexed (pemetrexed), tegafur (tegafur), uracil (uracil), uracil mustard (uracilmustard), fludarabine (fludarabine), gemcitabine (gemcitabine), capecitabine (capecitabine), mercaptopurine (mercaptoprine), cladribine (cladribine), thioguanine (thioguanine), methotrexate (methotrexate), pentostatin (pentostatin), hydroxyurea (hydroxyurea), or folic acid; antitumor antibiotics that interfere with DNA by inhibiting the action of enzymes and mitosis or altering cell membranes, such as pleomycin (phleomycin), bleomycin (bleomycin) or its derivatives or salts, CHPP, BZPP, MTPP, BAPP, libomycin (liblomycin), rifamycin (rifamycin), actinomycin (actinomycin), methacycline (adramycin), doxorubicin, epirubicin, pyrarubicin, daunorubicin, mitomycin; plant anti-cancer drugs derived from animal and plant components: such as paclitaxel (paclitaxel) [ taxol ], docetaxel (docetaxel) or taxotere (taxotere); vinca alkaloids, such as navelbine, vinblastine, vincristine, vindesine or vinorelbine; tropolone alkaloids such as colchicine (colchicine) or derivatives thereof; macrolides such as symptomatic denatols (macrolides); ansamitocins (ansamitocins) or rhizomycins (rhizoxins); antimitotic peptides, such as phomopsin or dolastatin; epipodophyllotoxin or podophyllotoxin derivatives such as etoposide (etoposide) or teniposide (teniposide); naracin (steganacin); antimitotic carbamate derivatives such as combretastatin (combretastatin) or amphetamine (amphetamine); procarbazine (procarbazine); camptothecin (camptothecin), such as irinotecan (irinotecan) (irinotecan hydrochloride and sorbitol injection camptosar) or topotecan (topotecan); hormonal antineoplastics, such as progesterone or estrogens, such as estramustine (T-66) or megestrol (megestrol); antiandrogens such as flutamide (flutamide), casodex (casodex), nilutamide (andron), or cyproterone acetate (cyproterone acetate); aromatase inhibitors, such as aminoglutethimide, anastrozole, formestan or letrozole; GHrH analogues, such as leuprorelin (leuprorelin), buserelin (buserelin), goserelin (goserelin) or triptorelin (triptorelin); antiestrogens such as tamoxifen (tamoxifen) or its citrate, droloxifene (droloxifene), troloxifene (trioxifene), raloxifene (raloxifene) or indoloxifene (zindoxifene); 17 β -estradiol derivatives, such as ICI 164, 384 or ICI 182, 780, aminoglutethimide (aminoglutethimide), formestan (formestan), fadrozole (fadrozole), finasteride (finasteride), ketoconazole (ketoconazole); LH-RH antagonists such as leuprolide acetate (leuprolide); steroids such as prednisone (prednisone), prednisolone (prednisone), methylprednisolone (methylprednisone), dexamethasone (dexamethasone), budesonide (budesonide), flucolone (fluocortolone) or triamcinolone (triamcinolone); other miscellaneous anti-neoplastic agents, such as proteasome inhibitors, e.g., bortezomib (bortezomib); enzymes, such as asparaginase, pegylated asparaginase (pegaspartase) or thymidine phosphorylase inhibitors; interferons, such as interferon beta; interleukins, such as IL-10 or IL-12; anti-TNF α antibodies, such as etanercept (etanercept); and targeted anti-tumor drugs, such as VEGF receptor small molecule antagonists, such as Watalanib (vatalanib), SU-5416, SU-6668, SU-11248, SU-14813, AZD-6474, EGFR/HER2 antagonists, such as CI-1033 or GW-2016; EGFR antagonists, such as Iressa [ gefitinib (gefitinib) ZD-1839], Tarceva (tarceva), PKI-166, EKB-569, HKI-272 or herceptin (herceptin); and antibodies targeting cancer cell surface molecules, such as apraclizumab (apolizumab) or 1D09C3, and the like.

The invention proves that DMSO has no protective effect on tumor cells, but the sulfoxide compound (preferably DMSO) can effectively prevent radioactive nervous system diseases generated by tumor radiotherapy, and has good protective effect on important nerve cells such as hippocampal neural stem cells and hippocampal neuron precursor cells. The sulfoxide compounds (preferably DMSO) can significantly improve behavioral symptoms related to radioactive nervous system diseases, such as memory impairment, memory deficiency, cognitive disorder, recognition/learning ability impairment, etc. Furthermore, DMSO was administered prophylactically over a wide time window, and it was expected that the prophylactic effect would be achieved with administration 4 hours prior to irradiation. DMSO has low potential toxic and side effects in the effective dose range of preventive administration.

Examples

Materials and methods:

1.1 materials, reagents and instruments

1.1.1 Experimental animals

The weight of an SPF grade C57BL/6J male mouse is 22-24 g; female SD rats, 6-8 weeks old, weighing 180 + -20 g, were purchased from Beijing Huafukang Biotech GmbH, Inc., animal quality certification number: SCXK (Jing) 2009-. All animals were housed in SPF grade barrier animal houses in cages of 5 animals each, sterilized water and maintained feed for free feeding. Keeping the daily light on for 12h and light off for 12h, and keeping the qualification number of the animal raising facility: SYXK- (military) -2007-. Mice were subjected to an adaptive post-feeding grouping experiment for about 7 days.

1.1.2 Primary reagents

(1) DMSO (south kyoton kangmanlin pharmaceutical science and technology ltd): 0.9 percent of medical normal saline is dissolved into specific concentration and is prepared as before use.

(2) Formaldehyde solution: 80ml of formaldehyde is taken from the measuring cylinder and added into 720ml of distilled water to prepare 10 percent formaldehyde solution.

(3) 5% goat serum blocking solution: blocking with normal sheep serum stock solution (ZLI-9021, product number) 0.5mL, adding 1 × TBST solution 9.5mL, mixing well, and storing at-20 deg.C.

(4) Horse Radish Peroxidase (HRP) -labeled goat anti-rabbit (Beijing China fir Jinqiao Biotechnology Co., Ltd., Cat. No. PV-6001).

(5) Horseradish peroxidase (HRP) -labeled goat anti-mouse (beijing sequoia kimoqiao biotechnology limited, cat # PV-6002).

(6) DAB kit 20 × (Beijing China fir Jinqiao Biotechnology Co., Ltd., Cat. number ZLI-9018).

(7) EDTA antigen retrieval solution pH9.0 (Beijing China fir Jinqiao biotechnology, Inc., Cat. number ZLI-9068): when in use, the product is diluted by 1:50 distilled water.

(8) 10% goat serum blocking solution: blocking with 1mL of normal sheep serum stock solution (Beijing Zhonghua Jinqiao Biotechnology Co., Ltd., product No. ZLI-9021), adding 9mL of 1 × TBST solution, mixing well, and storing at-20 deg.C.

1.1.3 instruments and consumables

X-ray machines (Rad Source Technologies, model RS-2000, USA).

An automatic dehydrator for biological tissue (Kedi instruments Co., Ltd., Jinhua, Zhejiang, model KD-TS 1A).

Biological tissue freezing embedding machine (Kedi instruments and equipments Co., Ltd., Jinhua, Zhejiang, model KD-BM, BL).

Computer biological tissue spreading machine (Kedi instruments Co., Ltd., Jinhua, Zhejiang, model KD-P).

Computer biological tissue baking machine (Kedi instruments Co., Ltd., Jinhua, Zhejiang, model KD-P).

Freezing microtome (Leica, Germany, CM3050S)

A rotary microtome (Leica, Germany, type RM 2235).

Upright fluorescence microscope (Leica, Germany, model DB 4000).

Digital cameras (canon, japan, model).

1.2 Experimental methods

1.2.1 Pre-irradiation preparation

One week before irradiation, the seeds are treatedThe experimental animals are raised in 5 cages in the center of SPF experimental animals, and the standard feed is added⁶⁰Feeding after sterilizing by CO gamma irradiation. Two days prior to irradiation, the experimental animals were weighed and randomly grouped according to the experimental design.

1.2.2 irradiation conditions

Before irradiation, a mouse is anesthetized by 1% pentobarbital sodium (0.2 ml/mouse, intraperitoneal injection), the mouse is placed in a self-made irradiation box in a supine position, a lead plate is placed below the neck of the mouse to completely shield other parts of the mouse, the tongue of the mouse is pulled out of an outlet, the irradiation conditions are that the voltage is 220kv, the current is 25mA, the dose rate is 1.379Gy/min, and the irradiation dose per time is respectively 5, 10 and 12 Gy. Each time, 7 mice were irradiated.

After the rats are anesthetized by intraperitoneal injection of 1% sodium pentobarbital (40mg/kg), the rats are fixed in a self-made irradiation box in a prone mode, and the neck marrow C2-T2 area of the rats is shielded and irradiated by lead plates, wherein the irradiation range is 2.0 multiplied by 1.0 cm. The irradiation conditions were voltage 220kv, current 25mA, dose rate 1.379Gy/min, and irradiation dose 22 Gy.

1.2.3 brain histopathological Observation

(1) BrdU intraperitoneal injection marker

BrdU (5mg/mL) was dissolved in saline, and the mice were intraperitoneally injected with BrdU solution (100mg/kg) twice at intervals of 8h 4 weeks after the injection, and the mice were sacrificed 16h after the second administration, and brain tissue was removed.

(2) Preparation of brain tissue material and section

The anesthetized mice were intraperitoneally injected with 0.2ml of 0.1% pentobarbital sodium, the chest was opened to expose the heart, the cut venous infusion needle was inserted into the left ventricle, the right auricle was opened, and heart perfusion was performed sequentially with PBS containing 1u/ml heparin and 2% PFA (paraformaldehyde, pH 7.4). After the perfusion is finished, the head is broken, the brain is taken, the brain tissue is placed into 2% PFA for room temperature fixation for 4 hours, then the brain tissue is sequentially placed into 10%, 20% and 30% sucrose solutions and placed at 4 ℃ for gradient dehydration (the sucrose solutions are prepared by PBS), and the brain tissue is placed for 24 hours respectively. The brains were then removed from the 30% sucrose solution and rinsed clean with PBS, then blotted dry with filter paper, and subsequently sectioned for embedding.

(3) Preparation of frozen sections

Continuously slicing the hippocampus by using a coronal section of a freezing microtome, wherein the thickness of brain slices is 30 μm, taking one brain slice every 150 μm, and storing the brain slice in an anti-freezing solution for later use.

(4) Immunohistochemistry

Rewarming the brain slice for 30min, washing with PBS 3 times;

3％H ₂O ₂blocking for 10min, washing 3 times with PBS.

Formamide, water bath at 65 ℃ for 40min for antigen retrieval, and PBS washing for 3 times;

2mol/L hydrochloric acid, water bath at 37 ℃ for 40min to untie DNA double chains, and washing for 3 times by PBS;

repairing with 0.01mmol/L Tris-HCL, and washing with PBS for 3 times;

primary antibody incubation: BrdU primary antibody (1:400), DCX primary antibody (1: 80), 4 ℃ refrigerator storage.

And (3) secondary antibody incubation: a Goat-anti-rat IgG (1:400) secondary antibody, incubating for 2h at room temperature, and washing for 3 times by PBS;

dripping ABC reagent (1:400) tertiary antibody, incubating for 2h at room temperature, and washing for 3 times with PBS;

DAB color development, hematoxylin nucleus staining;

dehydrating, and sealing.

(5) Cell counting

And (3) observing and photographing under a Leica DB4000B microscope, randomly selecting 3 discontinuous brain slices for each mouse to count cells, and respectively counting the BrdU positive cells and DCX positive cells under the microscope in the lower layer of the dentate gyropartical cells of the mouse.

1.2.4 histopathological Observation of the spinal cord

After the rats were anesthetized with 1% pentobarbital sodium (40mg/kg) by intraperitoneal injection, the chest was opened to expose the heart, the cut venous infusion needle was inserted into the left ventricle, the right auricle was opened, and heart perfusion was performed sequentially with PBS containing 1u/ml heparin and 2% PFA (paraformaldehyde, pH 7.4). After the perfusion is finished, taking out 2cm of the irradiated spinal cord, fixing in 4% paraformaldehyde for 7 days, dehydrating, embedding in paraffin, slicing, dyeing with Nissl dye liquor (Biyun day) for 10 minutes, and dehydrating and sealing after water washing. Neurons with diameters greater than 20 μm in the stained sections were counted under an optical microscope and 3 sections were analyzed per rat.

1.2.5 New object discrimination experiment

New object recognition experiments are commonly used for the assessment of learning and memory functions of animals. The experimental device is a square black opaque organic glass test box (45cm multiplied by 15cm), a camera is placed above the box body, and the motion of the camera is tracked and recorded. 2d before testing, the mice were placed in the experimental field and acclimated for 5min, 1 mouse at a time, 2 times daily. On the day of testing, the mice were placed in the experimental arena and allowed to explore freely for 3min, then removed and 2 identical objects (a1, a2) were placed 15cm apart from the edge of the arena, 2 at 15cm intervals. Mice were placed in the field facing the wall at equal distances from the two objects, and the time to explore the two objects was recorded within 10min (tA1 and tA2), respectively. After 1h, A2 was replaced by a new object B, the mice were placed again and the time to explore both objects (tA and tB) was recorded within 10min, respectively. After 24h, object B was changed to a new object C, the mice were placed again and the time to explore both objects (tA and tC) was recorded within 5min, respectively. The discrimination coefficients for the new objects are calculated separately.

The discrimination coefficient calculation formula is that a new object discrimination index (1h) is calculated as (tB-tA)/(tA + tB); the new object discrimination index (24h) ═ tC-tA)/(tA + tC) was calculated.

1.2.6Y maze spontaneous alternation experiment

The Y maze spontaneous alternation experiment is a commonly used test method for detecting the working memory of the rodent in the short-term space. The Y-maze consists of three identical arms which are 120 degrees apart and are trisected radially. During the experiment, the mouse is placed at the junction of the three arms, the mouse freely enters and exits the three arms, and the sequence of the mouse entering each arm and the total times of entering each arm within 8min are recorded. The spontaneous activity of all mice was recorded by a camera above the uterus. After each mouse experiment was completed and before the next mouse was placed in the maze, the device was wiped with a 75% alcohol cotton ball to reduce the odor interference to the experiment. The entry of a mouse limb into one arm was recorded, otherwise, the arm was considered as not entered. The correct alternation reaction was recorded as the correct alternation times by the continuous entry into three different arms, and the spontaneous alternation reaction rate was equal to the correct alternation times/(total arm entry times-2) × 100%. The spontaneous alternation response rate reflects the working memory capacity of the mice.

1.2.6 statistical treatment

Data results are expressed in x ± s and plotted using the GraphPad Prism6 software package. Statistical analysis of the data was performed using statistical software SPSS17.0(SPSS inc., Chicago), with P <0.05 accounting for the differences to be statistically significant.

Example 1 protective Effect of DMSO on proliferation of 5Gy head-irradiated mouse Hippocampus neural Stem cells

1. Grouping experiments:

12 male C57 mice were randomly drawn and divided into the following 4 groups:

non-irradiated control group (Non-IR): 4, only one of the active ingredients is added;

irradiation control group (Vehicle): gavage with 0.9% physiological saline, 0.2ml each, 4 pieces;

DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each 0.2ml, the dose is 10g/kg, 4 patients are administered 1 hour before irradiation;

irradiation dose: 5.0Gy head fraction irradiation.

2. The experimental method comprises the following steps:

see section 1.2 of the materials and methods section

3. The experimental results are as follows:

radiotherapy is one of the effective measures for treating craniocerebral tumor or preventing nervous system leukemia. The radiation can induce tumor cell death and damage to the nerve tissue surrounding the tumor. The neural stem cells in the brains of adult mice are mainly concentrated in the hippocampal dentate gyrus subvolume zone (SGZ) and the ventricular subvolume zone (SVZ) of the brain, and are most sensitive to radiation injury.

To understand the protective effect of DMSO on radioactive brain injury, the effect of DMSO prophylactic administration on proliferation of 5Gy irradiated mouse hippocampal neural stem cells was first observed using the BrdU incorporation method. Mice were irradiated to 5Gy and were injected intraperitoneally twice with BrdU solution (100mg/kg) 4 weeks after the second dose, at 8h intervals, sacrificed 16h after the second dose, brain tissue was removed, and BrdU immunohistochemical staining was performed.

The results showed that hippocampal BrdU positive cells were mainly distributed in the lower layer of granular cells in the dentate gyrus, the cytoplasm or nucleus was tan or yellowish, and the cells were scattered, as shown in fig. 1. As shown in Table 1, compared with the non-irradiated control group, the number of BrdU positive cells in the hippocampal region of the 5 Gy-irradiated mice is reduced, and the number of BrdU positive cells in the hippocampal region is obviously increased after DMSO (dimethyl sulfoxide) preventive administration compared with the irradiated control group, which indicates that DMSO (dimethyl sulfoxide) preventive administration has a protective effect on proliferation of hippocampal neural stem cells of the 5 Gy-irradiated mice.

TABLE 1

P < 0.01.

Example 2 protective Effect of DMSO on proliferation of 10Gy head-irradiated mouse Hippocampus neural Stem cells

1. Grouping experiments:

12 male C57 mice were randomly drawn and divided into the following 3 groups:

non-irradiated control group (Non-IR): 4, only one of the active ingredients is added;

irradiation control group (Vehicle): gavage with 0.9% physiological saline, 0.2ml each, 4 pieces;

DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each 0.2ml, the dose is 10g/kg, 4 patients are administered 1 hour before irradiation;

irradiation dose: 10.0Gy head fraction irradiation.

2. The experimental method comprises the following steps:

see section 1.2 of the materials and methods section

3. The experimental results are as follows:

to understand the protective effect of DMSO on radioactive brain injury after exposure to larger doses, the effect of DMSO prophylactic administration on proliferation of 10Gy irradiated mouse hippocampal neural stem cells was observed. Mice were irradiated with 10Gy and were injected intraperitoneally twice with BrdU solution (100mg/kg) 4 weeks after the second dose, at 8h intervals, sacrificed 16h after the second dose, brain tissue was removed, and BrdU immunohistochemical staining was performed.

As shown in FIG. 2 and Table 2, compared with the non-irradiated control group, the number of BrdU positive cells in the hippocampal region of the mice irradiated with 10Gy is more obviously reduced, and the number of BrdU positive cells in the hippocampal region of the mice after DMSO (dimethyl sulfoxide) prophylactic administration is obviously increased compared with the control group, which indicates that the DMSO prophylactic administration also has a remarkable protective effect on the proliferation of the neural stem cells in the hippocampal region of the mice irradiated with a larger dose of 10 Gy.

TABLE 2

P < 0.01.

Example 3 protective Effect of DMSO on 10Gy head-irradiated murine hippocampal precursor cell injury

1. Grouping experiments:

12 male C57 mice were randomly drawn and divided into the following 3 groups:

non-irradiated control group (Non-IR): 4, only one of the active ingredients is added;

irradiation control group (Vehicle): gavage with 0.9% physiological saline, 0.2ml each, 4 pieces;

DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each 0.2ml, the dose is 10g/kg, 4 patients are administered 1 hour before irradiation;

irradiation dose: 10.0Gy head fraction irradiation.

2. The experimental method comprises the following steps:

see section 1.2 of the materials and methods section

3. The experimental results are as follows:

there are neural stem cells in the adult mammalian brain that can differentiate into neurons or glia. Microtubule-associated protein, doubelocotrin (dcx), is expressed in neurons in the mitotic phase and early post-mitotic daughter neurons, and can be used as a marker of neuronal precursor cells to study proliferation and migration of neuronal precursor cells. The protective effect of DMSO prophylactic administration on hippocampal neuronal precursor cells was observed using DCX immunohistochemical staining.

As shown in Table 3, after 4 weeks of 10Gy head irradiation, the number of DCX positive neuron precursor cells in the hippocampus is obviously reduced compared with that in the non-irradiated group, and after DMSO is administered before irradiation, the number of DCX positive neuron precursor cells in the hippocampus is obviously increased, which indicates that DMSO prophylactic administration can reduce the damage of the neuron precursor cells caused by radiation.

TABLE 3

P < 0.01.

Example 4 DMSO prophylactic administration mitigates impairment of neoformation recognition Capacity in 10Gy head-irradiated mice

1. Grouping experiments:

28 male C57 mice were randomly drawn and divided into the following 4 groups:

non-irradiated control group (Non-IR): gavage with 0.9% physiological saline, 0.2ml each, 7 pieces;

non-irradiated DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each is 0.2ml, the dose is 10g/kg, and 7 patients are administered;

irradiation control group (Vehicle): gavage with 0.9% physiological saline, 0.2ml each, 7 pieces;

irradiation of DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each 0.2ml, the dose is 10g/kg, 7 patients are administered 1 hour before irradiation;

irradiation dose: 10.0Gy head fraction irradiation.

2. The experimental method comprises the following steps:

see section 1.2 of the materials and methods section

3. The experimental results are as follows:

the hippocampus is located between the thalamus and the medial temporal lobe, and is part of the limbic system, and is responsible for long-term memory storage, conversion, orientation, and other functions. After the irradiation of the mice, the proliferation capacity of the hippocampal neural stem cells is reduced, and the number of neuron precursor cells is reduced, so that the learning and memory capacity of the irradiated mice is possibly damaged.

The new object recognition test was carried out 4 weeks after the 10Gy head irradiation of the mice, and it was found that the new object recognition index at 1 hour intervals and the new object recognition index at 24 hours intervals were significantly reduced compared to the non-irradiated mice, the new object recognition index at 1 hour and 24 hours of the mice in the prophylactic group before DMSO irradiation was significantly improved, and DMSO had no effect on the new object recognition index of the non-irradiated mice (table 4). The result shows that the DMSO prophylactic administration can obviously reduce the damage of 10Gy head-irradiated mice on the recognition capability of the new object.

TABLE 4

P < 0.01.

Example 5 DMSO prophylactic administration mitigates impairment of working memory Capacity in 10Gy head-irradiated mice

1. Grouping experiments:

28 male C57 mice were randomly drawn and divided into the following 4 groups:

non-irradiated control group (Non-IR): gavage with 0.9% physiological saline, 0.2ml each, 7 pieces;

non-irradiated DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each is 0.2ml, the dose is 10g/kg, and 7 patients are administered;

irradiation control group (Vehicle): gavage with 0.9% physiological saline, 0.2ml each, 7 pieces;

irradiation of DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each 0.2ml, the dose is 10g/kg, 7 patients are administered 1 hour before irradiation;

irradiation dose: 10.0Gy head fraction irradiation.

2. The experimental method comprises the following steps:

see section 1.2 of the materials and methods section

3. The experimental results are as follows:

a Y maze spontaneous alternation experiment is carried out 4 weeks after the mice are irradiated, and the spontaneous alternation reaction rate of the 10Gy head-irradiated mice is obviously reduced compared with that of the non-irradiated mice, the spontaneous alternation reaction rate of the mice in a preventive administration group before DMSO irradiation is obviously improved, and the spontaneous alternation reaction rate of the DMSO on the non-irradiated mice is not obviously influenced. The results show that DMSO prophylactic administration can significantly reduce the impairment of working memory capacity in 10Gy head-irradiated mice (table 5).

TABLE 5

P < 0.01.

Example 6 protective Effect of DMSO prophylactic administration on 22Gy cervical irradiation rat nerve cell injury

1. Grouping experiments:

12 male C57 rats were randomly drawn and divided into the following 3 groups:

non-irradiated control group (Non-IR): 4, only one of the active ingredients is added;

irradiation control group (Vehicle): gavage with 0.9% physiological saline, 0.2ml each, 4 pieces;

DMSO group: the administration is carried out by stomach irrigation with 50% DMSO physiological saline, each 0.2ml, the dose is 10g/kg, 4 patients are administered 1 hour before irradiation;

irradiation dose: 22.0Gy cervical vertebrae local irradiation.

2. The experimental method comprises the following steps:

see section 1.2 of the materials and methods section

3. The experimental results are as follows:

the change of the number of spinal cord neurons 2 months after the 22.0Gy cervical vertebra local irradiation rat is irradiated is observed by applying a Nissl staining method, the number of the spinal cord neurons in the irradiated area is obviously reduced compared with that in the non-irradiated rat, and the number of the neurons positive to the Nissl in the irradiated area is obviously increased after DMSO is administered before irradiation, which indicates that the DMSO preventive administration can reduce the damage of the spinal cord neuron cells caused by rays (Table 6).

TABLE 6

P < 0.01.

Example 7 DMSO No radioprotection of head and neck tumor cells

1 materials and methods

1.1 materials and reagents

1.1.1 Experimental animals

The male BALB/C nude mouse is 22-24 g in weight, purchased from Beijing Huafukang Biotech GmbH, and has an animal quality certification number: SCXK (Jing) 2009-. All mice are delivered to an SPF-level barrier animal house in the center of laboratory animals of military medical academy of sciences by using a clean transport case and then are raised in cages, 5 mice are raised in each cage, sterilized water and feed are maintained for free feeding. Keeping the daily light on for 12h and light off for 12h, and keeping the qualification number of the animal raising facility: SYXK- (military) -2007-. Mice were subjected to an adaptive post-feeding grouping experiment for about 7 days.

1.1.2 cell lines: cal27 cells were purchased from the China center for type culture Collection (cell bank) of the institute of Life sciences, Wuhan university and were stored in the laboratory.

1.1.3 reagents and cell culture related reagents: DMSO (carnoka chemical biology, china, cat # D3855), MEM medium (beijing chinese michael, cat # 10010), Fetal Bovine Serum (FBS) (ExCell Bio, china, cat # FSP500), 0.25% trypsin digest (ExCell Bio, china, cat # 03.13005 a).

1.2 methods

The prepared suspension of human tongue squamous carcinoma Cal27 cells was inoculated into the right neck of nude mice with a 1mL syringe, and each injection was injected with 0.2mL (cell density 1X 10)⁶/mL), 20 nude mice were co-inoculated. After inoculation, the nude mice were returned to the super-clean laminar flow rack and raised as before.

When the tumor diameter of a nude mouse reaches 5mm, DMSO (6g/kg) or equivalent physiological saline is administered to the abdominal cavity of the tumor-bearing mouse, and after 1 hour, the nude mouse receives 12Gy of head and neck local shielding irradiation. Two other groups of tumor-bearing mice received no irradiation after administration of an equal amount of DMSO (6g/kg) or physiological saline. The tumor size was recorded 1 time per 3d observation, and the tumor volume was measured and calculated according to the volume formula V ═ d₁×d ₁×d ₂) Tumor volume was calculated (/ 2(d1 for tumor minor diameter, d2 for longest diameter perpendicular to d 1), tumor growth curves were plotted, and nude mice were weighed with an electronic balance. At 30 days after irradiation, the nude mice were sacrificed, the tumor tissue was stripped off, and weighed on an electronic balance.

2 results

The body weight of the non-irradiated tumor-bearing mice slowly decreased with the increase of the tumor, and there was no significant difference between the DMSO group and the saline control group. Two groups of tumor-bearing mice receiving 12Gy head and neck irradiation have obviously reduced endosome weight average 6 days after irradiation, the body weight of the mice in an irradiation control group is always maintained at a lower level, the body weight slightly rises 25 days after irradiation, one mouse dies, the body weight of the mice in a DMSO group obviously rises 12 days after irradiation, and compared with the irradiation control group, the DMSO group has obvious difference, and the DMSO group also has a protective effect on the oral mucositis of the tumor-bearing mice caused by radiation (figure 3 a).

As can be seen from FIGS. 3b-d, there was no significant difference between the tumor volume and the tumor weight after 30 days of administration in the non-irradiated tumor-bearing mice when DMSO was administered, compared with the control group. The 12Gy head and neck local irradiation can obviously inhibit the tumor growth, compared with two groups of tumor-bearing mice which are not irradiated, the tumor volume and the tumor weight after 30 days are obviously reduced, but the DMSO prevention group has no obvious difference compared with the control group. These results indicate that the DMSO prophylactic administration has no effect on the efficacy of tumor radiotherapy.

Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

Claims (12)

1. Use of a sulphoxide compound in the manufacture of a medicament for the prevention of a radiologic neurological disease in a subject suffering from a tumour to be treated with radiation.
2. Use according to claim 1, wherein the sulphoxide compound is DMSO or tetramethylene sulphoxide, preferably DMSO.
3. The use of claim 1 or 2, wherein the radioactive nervous system disease comprises radioactive brain injury (including acute radioactive brain injury, early late radioactive brain injury, and late radioactive brain injury), radioactive spinal cord injury (including chronic progressive radioactive spinal cord injury, muscle-collapsing radioactive spinal cord injury, acute radioactive spinal cord injury, and transient radioactive spinal cord injury), and radioactive peripheral nerve injury (including nerve cell injury, nerve stem cell injury, neuronal precursor cell injury, hippocampal neuronal stem cell injury, hippocampal neuronal precursor cell injury, neurite injury, nerve fiber injury, and neurosynaptic injury).
4. Use according to claim 1 or 2, wherein the radiologic neurological disease is manifested by one or more symptoms selected from: memory impairment, memory deficit, cognitive disorder, cognitive impairment, and learning impairment.
5. The use according to any one of claims 1 to 4, wherein the subject having a tumor is a subject having one or more tumors selected from the group consisting of: tumors of the digestive system, such as oral cancer, tonsil cancer, esophageal cancer, rectal cancer, colon cancer, liver cancer, pancreatic cancer, gastric cancer, etc.; tumors of the respiratory system, such as nasopharyngeal carcinoma, cancer of maxillary sinus, laryngeal carcinoma, bronchial cancer, lung cancer, etc.; genitourinary system tumor such as renal clear cell carcinoma, bladder cancer, nephroblastoma, testicular tumor, cervical cancer, uterine corpus cancer, etc.; breast cancer; tumors of the nervous system, such as brain tumor, medulloblastoma, neuroblastoma, pituitary tumor, etc.; malignant tumors of skin and soft tissue, such as penile cancer, lip cancer, melanoma, sarcoma; malignant tumor of bone, such as osteosarcoma, Ewing's tumor, etc.; and lymphoid neoplasms;

   preferably, the subject suffering from a tumor is a subject suffering from one or more of the following tumors: a subject suffering from a nervous system tumor, preferably a human suffering from a nervous system tumor; and subjects suffering from head and neck tumors, preferably humans suffering from head and neck tumors;

   preferably, the head and neck tumour is selected from one or more of: brain, medulloblastoma, neuroblastoma, and pituitary tumor;

   preferably, the head and neck tumour is selected from one or more of: tumors of the neck (e.g., thyroid tumors), otorhinolaryngological tumors (e.g., nasopharyngeal carcinoma, laryngeal carcinoma, or paranasal sinus cancer), and oromaxillofacial tumors (e.g., oral cancers, including tongue, gum, buccal, maxillary sinus, and tonsillar cancers).
6. Use according to any one of claims 1 to 5, wherein the subject suffering from a tumor is a subject of a head and neck tumor that will receive radiation therapy within 8 hours, preferably within 4 hours, more preferably within 2 hours, most preferably within 1 hour.
7. The use according to any one of claims 1 to 6, wherein the medicament is in a dosage form suitable for a mode of administration selected from one of: intravenous administration, oromucosal administration, transdermal administration and oral administration.
8. The use according to any one of claims 1 to 7, wherein the medicament is in a dosage form selected from the group consisting of: infusion solution, injection, collutory, pellicle, aerosol, tablet, adhesive, patch, sublingual agent, disintegrating agent, stick, powder, ointment, gel, water solution, suspension and capsule.
9. Use according to any one of claims 1 to 8, wherein the medicament is administered in a dose of 0.1 to 10.0g of sulphoxide, preferably 0.2 to 5.0g of sulphoxide, more preferably 0.4 to 2.0g of sulphoxide, most preferably 0.5 to 1.0g of sulphoxide per kg of body weight.
10. Use according to any one of claims 1 to 19, wherein the radiation therapy comprises a treatment with radiation selected from the group consisting of: alpha rays, beta rays, gamma rays, x rays, neutrons, electron rays, proton beams, particle beams, and combinations thereof; or the radiation therapy comprises external radiation therapy and internal radiation therapy; or the radiation therapy method includes conventional fractionated radiation therapy and one-time bolus radiation therapy.
11. Use according to any one of claims 1 to 10, wherein the radiation therapy has a radiation dose of 1 to 50Gy, more preferably 1 to 8Gy, most preferably 2 Gy.
12. The use of any one of claims 1 to 11, wherein the radiation therapy comprises the use of a radiosensitizer.

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