CN110917209A - Application of selenium-containing compound or selenium nano-grade in preparation of injection or microneedle of arthritis treatment drug - Google Patents
Application of selenium-containing compound or selenium nano-grade in preparation of injection or microneedle of arthritis treatment drug Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/04—Sulfur, selenium or tellurium; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
- A61K31/198—Alpha-aminoacids, e.g. alanine, edetic acids [EDTA]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
Abstract
The invention belongs to the technical field of biological medicines, and discloses an application of a selenium-containing compound or selenium nano in preparation of an injection or a microneedle for treating arthritis. The selenium-containing compound comprises a selenium-containing inorganic substance or a selenium-containing organic substance, the selenium-containing inorganic substance is sodium selenite, and the selenium-containing organic substance is one or two of selenomethionine and selenocysteine; the selenium nano comprises one or two of nano selenium and protein nano selenium. The injection is intracavity injection, local subcutaneous injection or local intramuscular injection. The invention provides a more rapid and safe treatment means for various arthralgia. Pharmacodynamic experiments prove that the medicine can obviously reduce the level of inflammatory cytokines in joint washing liquid of an arthritis model, has a local injection effect superior to the curative effect of a positive medicine, and has a good clinical application prospect.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to application of a selenium-containing compound or selenium nano-grade in preparation of an injection or microneedle for treating arthritis.
Background
Arthritis generally refers to inflammatory diseases occurring in joints and their surrounding tissues of the human body, and can be divided into tens of types. When various tissues of the locomotor system frequently move for a long time and intensively, when the fatigue limit is exceeded, such as long-term posture and movement are incorrect; or some injuries are caused by improper treatment although the injuries are acute; or arthritis can be caused by external factors such as improper exertion and external impact; the fatigue limit can be more easily reduced due to certain internal causes, such as degenerative changes, diseases, physical weakness, etc. of tissues. The clinical symptoms are long-term local pain, swelling, sensation of heat, partial sensation and dyskinesia. Among them, the most common Osteoarthritis (OA) is a degenerative joint disease, also known as degenerative joint disease, hypertrophic arthritis, senile arthritis, chondromalacia arthropathy, and the like, which is a clinically common disease. Currently, approximately 3.55 million people in the global population suffer from osteoarthritis, and in asia, 1 out of every 6 people. Knee osteoarthritis in particular has become a leading cause of motion affecting the elderly and leading to chronic disability. There are also various types such as rheumatoid osteoarthritis, gouty osteoarthritis, and the like. The etiology of the diseases is not completely clear, the clinical manifestations are similar, the red, swelling, heat, pain, dysfunction and joint deformity of the joints are taken as main symptoms, the commonly used anti-inflammatory drugs mainly comprise non-steroidal anti-inflammatory drugs, including ibuprofen, aspirin, diclofenac and the like, and the sitagliptin ointment taking diclofenac diethylamine as a main active ingredient is widely applied. However, the stimulation of gastrointestinal mucosa greatly limits the clinical application of these drugs, and there is a great need to develop alternative drugs with significant efficacy and low side effects.
Selenium is a necessary trace element for human body and plays an important role in physiological activities. Selenium has important biological functions of resisting oxidation, regulating immunity, antagonizing harmful heavy metals, etc. Selenium deficiency is related to more than 40 diseases, especially serious diseases such as AIDS, hepatitis B, liver cancer, keshan disease, Kaschin-Beck disease, cardiovascular and cerebrovascular diseases, etc. The proper selenium supplement can effectively prevent and control related diseases. Regarding the selenium intake standard of normal people, the World Health Organization (WHO) is specified to be 40-100 mug/d, the Chinese society for nutrition recommends to be 60-200 mug/d, and Canada is specified to be 98-224 mug/d. In more than 40 countries and regions of the world, about 72% of China, 2/3 people live in areas with low or low selenium. The nutrition survey result of province 13 in China shows that the daily intake of selenium for adults is only 26 mug, and the adults are seriously in a selenium deficiency state. Thus, the chinese population is generally deficient in selenium.
So far, the typical representation of selenium-containing inorganic substances is sodium selenite, which is stable in air, readily soluble in water, insoluble in ethanol; several pharmaceutical enterprises have been used at home and abroad to produce sodium selenite tablets for treating and preventing keshan disease and Kaschin-Beck disease which are endemic; CN1511541A/CN1273145C and CN1679622A/CN100340243C also disclose the use of tablets, granules, capsules and oral liquid prepared by taking sodium selenite as an active ingredient in the preparation of medicaments for treating osteoarthritis and rheumatoid arthritis; the CN100340243C and CN1273145C are orally taken sodium selenite 274 mu g/tablet (Na)2SeO3(ii) a The content of the converted Se is 125 mug/tablet), 1 tablet is taken each time, 2 times are taken each day (the total is about 250 mug/day), and 30 days are taken as a treatment course to take effect for treating the osteoarthritis (CN 100340243C); the rheumatoid arthritis takes 6 to 10 days (CN 1273145C); also, CN105434465B provides a method for preparing selenium-containing inorganic substance and zinc-containing inorganic substance according to the weight ratio of 1: 1-1: 5 range of the composition, and gel and milkThe ointment, the ointment or the patch is used for preventing and treating arthritis and is characterized in that the effect can be well observed only after the ointment is applied for 8 weeks, and the external application of sodium selenite has no remarkable statistical curative effect.
In 1985, Nuttal first proposed the hypothesis that colloidal state red elemental selenium has biological activity, but there is no evidence of research. In 1998, Zhang Jinsong and other people use reductant to reduce selenium compound in protein or polypeptide solution system to obtain nanometer red selenium dispersed in protein or polypeptide, and through separation and drying, nanometer selenium is obtained (CN 1059638C). Experimental research shows that the nano-selenium has better bioavailability, lower toxicity and higher bioactivity; such as resisting CCL4 induced acute liver injury, obviously inhibiting tumor volume, transferring, improving immunity and the like; the "selenium Wang" capsule is approved as a health food by the national ministry of health (Weishi Jian character 1998 No. 134). In 2002, Zhang Shengyi and the like have applied for patents of melatonin nano-selenium and a preparation method thereof (CN1415309), glucan nano-selenium and a preparation method thereof (CN1415241A), chitin nano-selenium and a preparation method thereof (CN1415310), amino acid nano-selenium and a preparation method thereof (CN1415308A), products of the products are all compounds consisting of certain template agents (such as melatonin, glucan, chitin and mixed amino acid) and red nano-selenium, and the products are required to be stored in a solid form. In 2003, Zhengwenjie and the like adopt a vitamin C direct reduction method to prepare sol-like nano elemental selenium (CN100496484C) which can be preserved in a liquid phase; in 2005, zhengwenjie et al reduced + selenium with valence 4 to selenium with valence 0 using the antioxidant property of phycocyanin itself to form composite nano-selenium (CN 1947792B); in 2007, Zhengwenjie et al constructed the preparation of the complex of amino acid and nano elemental selenium by using amino acid as a template (CN 101040869A). In 2018, the applicant constructs a novel preparation technology of protein-bound nano elemental selenium (CN108484715A) which is easier to store in a liquid phase and higher in safety based on protein self-assembly.
However, selenium is a trace beneficial element, the effectiveness and safety of selenium are always the first major problems to be balanced, and although oral administration of sodium selenite and long-term external application of sodium selenite and zinc-containing inorganic compositions can effectively relieve arthritis, the safety hazard still exists. In view of this, the preparation of the nano-selenium and the high-liquid-phase preservation protein nano-selenium with high activity and high safety provides firmer guarantee for the safe use of the selenium preparation. Most importantly, until now, the application of selenium-containing inorganic substances, selenium-containing organic substances, nano-selenium and protein nano-selenium in the direct treatment of arthritis and joint cavity parts to quickly relieve the symptoms of arthritis is not seen.
Disclosure of Invention
The invention aims to solve the defects and shortcomings of poor clinical drug curative effect, oral administration of drugs, stimulation of gastrointestinal mucosa to generate adverse reaction, long effective period of external application, potential safety hazard of excessive selenium element and the like in the prior art, and provides application of a selenium-containing compound or selenium nano in preparation of an injection or microneedle for treating arthritis.
The purpose of the invention is realized by the following technical scheme:
the application of selenium-containing compound or selenium nano-grade in preparing injection or microneedle for treating arthritis is provided.
The selenium-containing compound comprises a selenium-containing inorganic substance or a selenium-containing organic substance, the selenium-containing inorganic substance is sodium selenite, and the selenium-containing organic substance is one or two of selenomethionine and selenocysteine; the selenium nano comprises one or two of nano selenium and protein nano selenium.
The injection is intracavity injection, local subcutaneous injection or local intramuscular injection.
The intracavity injection is injected in the joint cavity; the local subcutaneous injection is injected subcutaneously at the joint part; the local intramuscular injection is injected intramuscularly at the joint site.
The concentration of the selenium element in the medicine is 0.001-1000 mu M.
The therapeutic dose of the medicine according to the selenium element is 0.001-100 mu g/time.
The medicine is prepared by taking a selenium-containing compound or selenium nano as a raw material medicine and adding pharmaceutically acceptable auxiliary materials or auxiliary components.
The pH range of the injection or the microneedle is 5.5-7.4.
The medicine for treating arthritis is a medicine for treating osteoarthritis, rheumatic arthritis, rheumatoid arthritis, gouty arthritis, osteoarthritis of the greater bone, reactive arthritis, infectious arthritis, traumatic arthritis, carpal tunnel syndrome, bursitis or synovitis.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the invention prepares the selenium-containing compound or the selenium nano-grade into a local injection preparation, and provides a more rapid, direct and safe treatment means for various arthralgia; compared with oral administration or long-term external application, the selenium/zinc salt compound ointment can solve the safety problem of selenium accumulation caused by difficult transportation into inflammatory cells and long-term use of the traditional selenium/zinc salt compound ointment, and the invention can skillfully solve the problem. Pharmacodynamic experiments prove that the invention can obviously reduce the level of inflammatory cytokines in the joint washing liquid of an arthritis model, the treatment effect of the invention can even reach the curative effect better than that of a positive medicament, and mechanism researches find that the parallel iron death inhibitor or iron complexing agent has similar biological effect with the iron death inhibitor or iron complexing agent, which indicates that the invention is also one of iron death inhibitor molecules. Therefore, the invention has good clinical application prospect.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1 preparation of comparative selenium gel (SE):
prescription: 0.5g of sodium selenite, 94010 g of carbomer, 75g of glycerol, 802g of polysorbate, 13.5g of triethanolamine and distilled water added to 1000 g.
The process comprises the following steps: mixing carbomer 940, polysorbate 80 and 300g of distilled water, dissolving triethanolamine in 100g of distilled water, adding the above solution, stirring, adding glycerol, and stirring to obtain matrix. Dissolving zinc sulfate and sodium selenite in 250g distilled water, adding into the above matrix after dissolving completely, adding water to full volume, stirring well, and removing bubbles to obtain selenium gel SE.
Example 2 preparation of nano-selenium (NSe):
referring to the patent application (publication No. CN1059638C) and the literature (Life Sciences 2004; 75: 237-.
Example 3 preparation of protein-bound nanoselenium (PSe):
referring to example 1 of the applicant's patent application (publication No. CN108484715A), human serum albumin-bound nano-selenium (PSe) was prepared, and the obtained protein-bound selenium nanoparticles PSe were uniform in nano-size, uniform in distribution, and about 30-40nm in average particle size.
Example 4 a method of preparing sodium selenite injection, the formulation is:
every 20L of sodium selenite injection is prepared, the addition amount of the edetate disodium is 2g, the addition amount of the sodium metabisulfite is 38g, the addition amount of the selenite is 100g, and the addition amount of the sodium bicarbonate is 2.49 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, sodium metabisulfite and sodium selenite into water for injection saturated by carbon dioxide, completely dissolving at 60 ℃, then adding sodium bicarbonate, adding water for injection or normal saline to a constant volume, and adjusting pH to 6.4 with citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not greater than 0.25 micrometer; then continuously introducing carbon dioxide gas into the filtrate after fine filtration, sealing under the condition of introducing carbon dioxide, and sterilizing at 115 deg.C for 20min to obtain selenium-containing injection.
Example 5a method of preparing sodium selenite injection, the formulation is:
every 20L of sodium selenite injection is prepared, the addition amount of the edetate disodium is 2g, the addition amount of the sodium metabisulfite is 38g, the addition amount of the selenite is 150g, the addition amount of the sodium hyaluronate is 200g, and the addition amount of the sodium bicarbonate is 2.49 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, sodium hyaluronate, sodium metabisulfite and sodium selenite into water for injection saturated by carbon dioxide, completely dissolving at 40 ℃, adding sodium bicarbonate, adding water for injection or normal saline to a constant volume, and adjusting pH to 7.2 with citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not greater than 0.25 micrometer; then continuously introducing carbon dioxide gas into the filtrate after fine filtration, sealing under the condition of introducing carbon dioxide, and sterilizing at 115 deg.C for 20min to obtain selenium-containing injection.
Example 6 a method of preparing sodium selenite injection, the formulation is:
every 20L of sodium selenite injection is prepared, the addition amount of the edetate disodium is 2g, the addition amount of the sodium metabisulfite is 38g, the addition amount of the selenite is 190g, the glucosamine hydrochloride is 2Kg, and the addition amount of the sodium bicarbonate is 2.49 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, glucosamine hydrochloride, sodium metabisulfite and sodium selenite into water for injection saturated by carbon dioxide, completely dissolving at 50 ℃, adding sodium bicarbonate, adding water for injection or normal saline to a constant volume, and adjusting pH to 6.9 with citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not greater than 0.25 micrometer; then continuously introducing carbon dioxide gas into the filtrate after fine filtration, sealing under the condition of introducing carbon dioxide, and sterilizing at 115 deg.C for 20min to obtain selenium-containing injection.
Example 7 a method of making selenomethionine injection, the formulation is:
every 20L of selenomethionine injection is prepared, the addition amount of disodium edetate is 2g, the addition amount of sodium metabisulfite is 38g, the addition amount of selenomethionine is 199g, and the addition amount of sodium bicarbonate is 2.49 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, glucosamine hydrochloride, sodium metabisulfite and selenomethionine into water for injection saturated by carbon dioxide, completely dissolving at 50 ℃, adding sodium bicarbonate, adding water for injection or normal saline to a constant volume, and adjusting pH to 6.9 with citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not greater than 0.25 micrometer; then continuously introducing carbon dioxide gas into the filtrate after fine filtration, sealing under the condition of introducing carbon dioxide, and sterilizing at 115 deg.C for 20min to obtain selenium-containing injection.
Example 8a method of making selenomethionine injection, the formulation is:
every 20L of selenomethionine injection is prepared, the addition amount of edetate disodium is 2g, the addition amount of sodium metabisulfite is 38g, the addition amount of selenomethionine is 170g, glucosamine hydrochloride is 2Kg, and the addition amount of sodium bicarbonate is 2.49 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, glucosamine hydrochloride, sodium metabisulfite and selenomethionine into water for injection saturated by carbon dioxide, completely dissolving at 50 ℃, adding sodium bicarbonate, adding water for injection or normal saline to a constant volume, and adjusting pH to 6.1 with citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not greater than 0.25 micrometer; then continuously introducing carbon dioxide gas into the filtrate after fine filtration, sealing under the condition of introducing carbon dioxide, and sterilizing at 115 deg.C for 20min to obtain selenium-containing injection.
Example 9a method of preparing a nano-selenium injection, the formulation is:
every 20L of nano-selenium injection is prepared, the addition amount of the disodium edetate is 2g, the addition amount of the sodium metabisulfite is 38g, the addition amount of the nano-selenium obtained in example 2 is 90g, and the addition amount of the sodium bicarbonate is 2.49 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, sodium metabisulfite and nano-selenium into water for injection saturated by carbon dioxide, completely dissolving at 50 ℃, then adding sodium bicarbonate, finally adding water for injection or normal saline to a constant volume, and adjusting pH to 6.5 by using citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not more than 0.22 micrometer; and continuously introducing carbon dioxide or nitrogen gas into the filtrate after fine filtration in a sterile environment, and filling and sealing under the condition of introducing the carbon dioxide or the nitrogen gas to obtain the selenium-containing injection.
Example 10 a method of preparing a nano-selenium injection, the formulation is:
every 20L of nano-selenium injection is prepared, the addition amount of the disodium edetate is 2g, the addition amount of the sodium metabisulfite is 38g, the addition amount of the nano-selenium obtained in example 2 is 100g, and the addition amount of the sodium bicarbonate is 2.19 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, sodium metabisulfite and nano-selenium into water for injection saturated by carbon dioxide, completely dissolving at 40 ℃, then adding sodium bicarbonate, finally adding water for injection or normal saline to a constant volume, and adjusting pH to 7.1 by using citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not more than 0.22 micrometer; and continuously introducing carbon dioxide or nitrogen gas into the filtrate after fine filtration in a sterile environment, and filling and sealing under the condition of introducing the carbon dioxide or the nitrogen gas to obtain the selenium-containing injection.
Example 11 preparation method of nano selenium protein injection, the formulation is:
every 20L of selenomethionine injection is prepared, the addition amount of edetate disodium is 2g, the addition amount of sodium metabisulfite is 38g, the addition amount of the protein nano-selenium obtained in example 3 is 85g, and the addition amount of sodium bicarbonate is 2.40 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, sodium metabisulfite and protein nano selenium into water for injection saturated by carbon dioxide, completely dissolving at 40 ℃, then adding sodium bicarbonate, finally adding water for injection or normal saline to a constant volume, and adjusting pH to 7.2 by using citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not more than 0.22 micrometer; and continuously introducing carbon dioxide or nitrogen gas into the filtrate after fine filtration in a sterile environment, and filling and sealing under the condition of introducing the carbon dioxide or the nitrogen gas to obtain the selenium-containing injection.
Example 12 a method for preparing a protein nano-selenium injection, the formulation is:
every 20L of selenomethionine injection is prepared, the addition amount of edetate disodium is 2g, the addition amount of sodium metabisulfite is 38g, the addition amount of the protein nano-selenium obtained in example 3 is 105g, and the addition amount of sodium bicarbonate is 2.41 Kg; the method comprises the following steps:
(1) sequentially adding disodium edetate, sodium metabisulfite and protein nano selenium into water for injection saturated by carbon dioxide, completely dissolving at 40 ℃, then adding sodium bicarbonate, finally adding water for injection or normal saline to a constant volume, and adjusting pH to 7.0 by using citric acid or sodium bicarbonate to obtain a mixed solution;
(2) adding medicinal activated carbon into the mixed solution obtained in the step (1), adding 0.2g of medicinal activated carbon into each liter of the mixed solution, stirring, filtering for decarbonization, and finely filtering the filtered filtrate by using an ultrafiltration membrane with the pore diameter not more than 0.22 micrometer; and continuously introducing carbon dioxide or nitrogen gas into the filtrate after fine filtration in a sterile environment, and filling and sealing under the condition of introducing the carbon dioxide or the nitrogen gas to obtain the selenium-containing injection.
EXAMPLE 13 quality testing of formulations
The selenium-containing injection prepared in the above examples 4 to 12 is subjected to content determination of active ingredients, sodium thiosulfate titration solution is adopted to titrate the content of selenium, or a concentrated nitric acid digestion method is adopted to carry out detection by ICP-MS, the content is 90.0% -110.0% of the marked amount, 5 samples are determined in each example, and the content of selenium in the samples in the examples 4 to 12 is found to be more than 97% of the marked content.
And (3) stability test: and (3) placing the sample in a stability test box, standing for 12 months under the conditions of 40 ℃ and 60% of humidity, and testing to ensure that the property, the content, the pH value and the sterile test of the product meet the requirements.
Long-term tests (25 ℃. + -. 2 ℃; RH 40%. + -. 5%): the test sample is placed flat under the conditions that the temperature is 25 +/-2 ℃ and the relative humidity is 40 +/-5%, and sampling detection is carried out after 18 months, so that the injection is transparent, the color is clear, and the pH value is changed within 0.2.
And (4) conclusion: the invention has qualified properties, content measurement and sterile experiment, and meets the relevant requirements of pharmacopoeia 2015 year edition of the people's republic of China.
Example 14 in vivo efficacy experiment one: rabbit osteoarthritis model
The effect of the drug on the content of the cytokines (TNF- α and IL-1 β) in the rabbit knee osteoarthritis model is examined through detecting conventional biomarkers of inflammation, such as cytokines and chemokines, such as TNF- α (tumor necrosis factor- α), NF-kB (transcription factor protein family) or IL (interleukin) and the like.
The material and the method are as follows: experimental materials: experimental animals healthy New Zealand white rabbits 190 animals, each half male and female, with a body weight of 2.6-3.0Kg, were provided by Schleik laboratory animals Co., Ltd, Hunan. Feeding the standard feed at room temperature of 15-25 ℃, normally illuminating every day, circulating air, and freely taking and drinking water. Experimental drugs: injectable formulations and external reference ointments prepared according to examples 4-12 example 1, positive control drugs were applied externally: a sertraline ointment; group of positive drugs for intracavitary injection: prednisolone acetate hormone injection (125mg/5 ml; 0.2 ml); ferrostatin-1 injection (5 mg/ml; 0.2 ml); dexrazoxane DXZ injection (125 mg/ml; 0.2 ml); necrostatin-1 injection (10 mg/mL; 0.2 mL); Z-VAD-FMK injection (10 mg/mL; 0.2 mL).
The experimental method comprises the following steps: animal grouping: 190 New Zealand white rabbits were divided into 19 groups by simple random sampling: blank group, model group (externally applied ointment matrix group and normal saline injection group), externally applied positive drug ointment control group, intracavity injection hormone group, externally applied gel ointment SE group, iron death inhibitor (Ferrostatin-1, Ferr-1) group, iron complexing agent Dexrazoxane (DXZ) group, programmed apoptosis inhibitor (Nerostatin-1, N-1) group, broad spectrum Caspase inhibitor Z-VAD-FMK group (Z-VAD-FMK), treatment group 9 group (respectively are selenium-containing injection prepared in examples 4-12), 10 of each group, weighing and marking.
The molding method comprises the following steps: the left knee joints of the experimental animals were fixed in the straightened position by the Vandman method in the other 18 groups except the blank group without molding. The fixation range is from 3cm below the ankle joint of the rabbit to 1.5cm below the groin, the ankle is dorsiflexed at 30-40 degrees, and the fixation time is 6 weeks; after fixing for 6 weeks, randomly killing 1 animal in each group, and taking the articular cartilage for pathological examination to observe the molding condition; after the molding is successful, the fixation is removed and the hair is shaved.
The administration condition is as follows: after each group successfully constructs the model, externally coating the control group with the nataline ointment with the thickness of about 2mm, and uniformly covering the surface of the joint of the model for 2 times a day; the treatment groups respectively inject the preparation prepared according to the embodiment 4-12 into the joint cavity, 100-200 ng/time (200 mu l), 1 time per week; group of positive drugs for intracavitary injection: 5.0 mg/time, 1 time per week; iron death inhibitor (Ferrostatin-1, Ferr-1) group, 1.0 mg/time, 1 time per week; dexrazoxane (dxdz), group of iron complexing agents, 25.0 mg/time, 1 time per week; apoptosis inhibitor (Necrostatin-1, N-1) group, 2.0 mg/time, 1 time per week; broad-spectrum caspase inhibitor Z-VAD-FMK group (Z-VAD-FMK), 2.0 mg/time, 1 time per week; external application gel ointment SE group the gel prepared according to comparative example 1, having a thickness of about 2mm, was applied to the surface of the molded joint evenly, 2 times a day. The external application model group is coated with vaseline with the thickness of about 2mm, and the model joints are uniformly covered 2 times per day. In the blank group, vaseline is applied on the surface of the right knee joint, the thickness is about 2mm, and 2 times per day; saline injection model the control group was injected intraluminally at a rate of 0.2 ml/time per week.
The detection method comprises injecting 30ml of air into ear vein after 1 week, killing rabbit rapidly, fixing on dissecting frame, disinfecting knee joint rapidly, taking about 0.5cm above the attachment point of patellar ligament, injecting 1.5ml of distilled water into joint cavity along outer edge of ligament, sucking joint liquid repeatedly for several times, extracting about 2ml of joint mixed liquid, centrifuging the joint mixed liquid for 10min at 4000r/min, sucking out about 0.3ml of supernatant, placing into test tube, storing in-20 deg.C refrigerator, and determining TNF- α and IL-1 β content in joint washing liquid according to the content in kit.
Experimental results experimental data are expressed as mean ± standard deviation (x ± s), and were compared two by two between groups using t or t' test, test level α ═ 0.05, and analyzed using SPSS 17.0 and Graphprism software.
TABLE 1 Effect of the inventive injection on the levels of cytokines TNF- α and IL-1 β in the synovial fluid of rabbit and knee osteoarthritis model (mean. + -. SD)
| Group of | Animal number (number) | TNF-α(ng/ml) | IL-1β(ng/ml) |
| Blank group | 10 | 1.64±0.26 | 0.28±0.07 |
| Model group/external application ointment base | 10 | 2.95±0.19### | 0.75±0.13### |
| Model group/intracavity physiological saline injection | 10 | 2.98±0.17### | 0.78±0.14### |
| Control group/external application of sertraline ointment | 10 | 2.90±0.17 | 0.73±0.09 |
| Control group/intracavity injection prednisolone acetate | 10 | 1.98±0.19*** | 0.60±0.07*** |
| External application/SE gel ointment group | 10 | 2.94±0.21 | 0.76±0.08 |
| Ferr-1 group of iron death inhibitors | 10 | 1.87±0.15*** | 0.39±0.07*** |
| Iron complexing agent dexrazoxane DXZ group | 10 | 1.98±0.19*** | 0.49±0.08*** |
| Apoptosis inhibitor N-1 group | 10 | 2.92±0.23 | 0.72±0.07 |
| Caspase inhibitors Z-VAD-FMK group | 10 | 2.93±0.19 | 0.71±0.06 |
| Example 4 | 10 | 2.11±0.16*** | 0.53±0.07*** |
| Example 5 | 10 | 2.15±0.19*** | 0.57±0.05*** |
| Example 6 | 10 | 2.18±0.14*** | 0.49±0.03*** |
| Example 7 | 10 | 2.05±0.11*** | 0.45±0.05*** |
| Example 8 | 10 | 2.03±0.16*** | 0.47±0.06*** |
| Example 9 | 10 | 1.85±0.15*** | 0.35±0.05*** |
| Example 10 | 10 | 1.83±0.13*** | 0.36±0.05*** |
| Example 11 | 10 | 1.68±0.14*** | 0.31±0.03*** |
| Example 12 | 10 | 1.70±0.17*** | 0.30±0.05*** |
# p <0.001 compared to blank; p < 0.05; p < 0.01; p <0.001 was compared to the corresponding model groups.
The contents of TNF- α and IL-1 β in the joint flushing fluid of a blank group are lower than those of a model group, and have significant difference (p is less than 0.01), which indicates that inflammatory cytokines in the joint fluid of the left knee and the knee of a rabbit are increased after modeling, the contents of TNF- α and IL-1 β in the joint flushing fluid of each treatment group are lower than those of the model control group, and the significant difference (p is less than 0.01) exists, but the significant difference does not exist between the programmed apoptosis inhibitor N-1 group and the Caspase inhibitor Z-VAD-FMK group, which indicates that the action mechanism of the injection is not significantly related to apoptosis, and experiments indicate that the selenium-containing injection can significantly reduce the level of inflammatory cytokines in the joint flushing fluid of an arthritis model, and the action mechanism of the injection is closely related to an iron death pathway.
In the same treatment period, the content of TNF- α and IL-1 β in joint washing liquid is not obviously different from that of a model group by singly using a selenium-containing gel or externally-applied positive medicine ointment group, which indicates that the anti-inflammatory effect of treating arthritis by singly using selenium gel or externally-applied positive medicine in a short time is poor, but only the selenium-containing injection and the intracavity injection hormone can quickly relieve the inflammation, and meanwhile, swelling and fading can be obviously observed within 6 hours after the joint cavity injection, and the selenium-containing injection is also prompted to achieve the ideal swelling and anti-inflammatory effect.
The above experimental results show that: the selenium-containing injection has a remarkable treatment effect on arthritis, while the SE gel ointment group is ineffective, particularly the nano-selenium injection and the protein nano-selenium injection have remarkable treatment effects, and the action mechanism of the selenium-containing injection is probably closely related to the iron death pathway.
Example 15 in vivo efficacy experiment two: rat gout model
Purpose of the experiment: the anti-gout effect of the medicine is evaluated through a rat induced gout model.
The method comprises the steps of preparing a medicine, namely prednisolone acetate (Chongqing medicine-free pharmaceutical Limited liability company), sodium urate crystals (HPLC is more than or equal to 99%, 20mg/mL of molding solution is prepared from normal saline or PBS for injection), an IL-1 β ELISA kit, medical normal saline for injection, and other analytically pure reagents, preparing a sodium urate solution, namely adding 5mL of 1mol/L NaOH and 800mg of sodium urate into 155mL of sterilized injection water for removing pyrogens under an aseptic condition, boiling to completely dissolve the sodium urate, naturally cooling and stirring, dripping 1mol/L of HCl to a pH value of 7.0, repeatedly centrifuging (3000r/min and 2min) after the solution is milky, cooling for 1h at 4 ℃, collecting crystals, storing at 4 ℃, baking supernatant, a beaker, a stirring rod and a centrifuge tube, weighing, preparing Phosphate Buffer Solution (PBS) in the phosphate buffer solution for preparing phosphate buffer solution for the crystals, preparing the phosphate buffer solution for later use, preparing the phosphate buffer solution for preparing phosphate buffer solution, preparing the phosphate buffer solution for later use, preparing the phosphate buffer solution, adding the phosphate buffer solution for the phosphate buffer solution, preparing the phosphate buffer solution, and the phosphate buffer solution for later, and the phosphate buffer solution for preparing the phosphate buffer solution for later use, wherein the phosphate solution is prepared for injection, the.
Experimental animals: SD male rats, body weight 200 ± 10g (provided by the hunan slaick experimental animal center); the seeds are raised indoors in cages at room temperature (20-22 ℃), the relative humidity is 35% -50%, and the seeds can be eaten and drunk freely. Three days after adaptive feeding, the divided dosing was started.
Animal grouping and modeling: 170 SD rats, randomly divided into 17 groups by weight, 10 rats per group, and each group was designated as a blank group, a model group, a positive drug hormone control group (prednisone acetate group, 0.4mg/300g ═ 1.32mg/kg of body weight, intra-articular injection), an external application gel ointment SE group (external application gel ointment SE group is externally applied with the gel prepared according to comparative example 1, the thickness is about 2mm, the gel uniformly covers the surface of the molded joint, 3 times per day), and an iron death inhibitor (Ferrostatin-1, Ferr-1) group (1.5mg/kg, single intra-articular injection); dexrazoxane (dxzz) group (15.0mg/kg, single intracavitary injection) iron complexing agent; apoptosis inhibitor (Necrostatin-1, N-1) group (6.0mg/kg, single intracavity injection); broad-spectrum caspase inhibitors Z-VAD-FMK group (Z-VAD-FMK; 6.0mg/kg, single intracavitary injection); treatment groups 9 (selenium-containing injections prepared in examples 4-12, respectively), the 17 animals mentioned above, were fed freely and had water; intraperitoneal injection of pentobarbital sodium (40mg/kg) for anesthesia, fixing a rat, bending a right ankle joint, locally sterilizing the periphery of the right ankle joint by using 3% iodine tincture and 75% alcohol, and inserting a needle between two apophysis by using a No. 6 injection needle, specifically inserting a needle into a gap between a dorsal median ankle joint of a right hind limb and a tibiofibula; except that the blank group of rats are injected with 0.05mL of physiological saline, the other groups of rats are injected with 0.05mL of sodium urate physiological saline suspension in the right joint cavity, and the occurrence of gouty arthritis is induced by taking the bulge of the opposite side of the joint capsule as an injection standard; from the basic cadence of the model: the sodium urate crystal can rapidly induce joint red swelling and dysfunction after being injected for 2h, the symptoms are obvious after 4h, the peak is 8-12h, the peak is relieved after 24h, and the symptom disappears after 5-6 d; after the injection of the sodium urate crystal in the joint cavity, obvious effusion and fiber exudation occur, a large amount of inflammatory cell infiltration mainly comprising neutrophils is generated, the tissues are seriously edematous, and the synovium and the surrounding tissues thereof are also infiltrated by a large amount of inflammatory cells. At 4h after model induction, the inventor further injected each group of drugs or physiological saline 0.05 mL.
Sample treatment and determination (1) degree of swelling of joints, namely, the thickness of the right ankle joint of a rat before and after inflammation is measured by a vernier caliper respectively, the thickness of the right ankle joint of the rat before and after inflammation is divided by the thickness before inflammation multiplied by 100% to be used as the swelling degree of gouty arthritis, (2) influence on IL-1 β in the joint and surrounding soft tissues of the rat, namely, after blood is taken after 48 hours of inflammation is caused, the animal is killed, the synovial membrane of the joint and the surrounding soft tissues are taken, the mass is weighed, homogenate is prepared in an ice bath, physiological saline is added according to the proportion of 1: 20, the mixture is centrifuged at 4000r/min for 5min, supernatant is taken and is subpackaged in Ep tubes, and is preserved in an ELISA refrigerator at the temperature of-80 ℃, and IL-1 β is quantitatively detected according to the specification of a double antibody sandwich method kit.
Experimental results experimental data are expressed as mean ± standard deviation (x ± s), and were compared two by two between groups using t or t' test, test level α ═ 0.05, and analyzed using SPSS 17.0 and Graphprism software.
TABLE 2 Effect of the pharmaceutical composition of the present invention on arthrocele in gout rats (mean. + -. SD)
# p <0.001 compared to blank; p < 0.05; p < 0.01; p <0.001 was compared to the corresponding model groups.
TABLE 3 Effect of the pharmaceutical compositions of the present invention on IL-1 β levels in arthritic tissues of gout rats (mean. + -. SD)
# p <0.001 compared to blank; p < 0.05; p < 0.01; p <0.001 was compared to the corresponding model groups.
The experimental result shows that, like the positive drug hormone, the selenium-containing compound, the selenium nano-grade, the iron death inhibitor Ferr-1 group and the iron complexing agent dexrazine DXZ group have the effects of remarkably improving the painful joint swelling of rats and reducing the IL-1 β level in tissues and present good biological effect relationship, while the programmed apoptosis inhibitor N-1 group, the Caspase inhibitor Z-VAD-FMK group and the SE gel ointment group are ineffective, which indicates that the programmed apoptosis inhibitor Z-VAD-FMK group and the SE gel ointment group are not closely related to the apoptosis, so that the intracavity injection of the selenium-containing compound and the selenium nano-grade have the remarkable effect of resisting the painful arthritis, the effect of protein combined with the nano-grade selenium is most obvious, and the action mechanism of the intracavity injection of the selenium-containing compound and the selenium nano-grade is probably closely related to.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (9)
1. The application of selenium-containing compound or selenium nano-grade in preparing injection or microneedle for treating arthritis is provided.
2. Use according to claim 1, characterized in that: the selenium-containing compound comprises a selenium-containing inorganic substance or a selenium-containing organic substance, the selenium-containing inorganic substance is sodium selenite, and the selenium-containing organic substance is one or two of selenomethionine and selenocysteine; the selenium nano comprises one or two of nano selenium and protein nano selenium.
3. Use according to claim 1, characterized in that: the injection is intracavity injection, local subcutaneous injection or local intramuscular injection.
4. Use according to claim 3, characterized in that: the intracavity injection is injected in the joint cavity; the local subcutaneous injection is injected subcutaneously at the joint part; the local intramuscular injection is injected intramuscularly at the joint site.
5. Use according to claim 1, characterized in that: the concentration of the selenium element in the medicine is 0.001-1000 mu M.
6. Use according to claim 1, characterized in that: the therapeutic dose of the medicine according to the selenium element is 0.001-100 mu g/time.
7. Use according to claim 1, characterized in that: the medicine is prepared by taking a selenium-containing compound or selenium nano as a raw material medicine and adding pharmaceutically acceptable auxiliary materials or auxiliary components.
8. Use according to claim 1, characterized in that: the pH range of the injection or the microneedle is 5.5-7.4.
9. Use according to claim 1, characterized in that: the medicine for treating arthritis is a medicine for treating osteoarthritis, rheumatic arthritis, rheumatoid arthritis, gouty arthritis, osteoarthritis of the greater bone, reactive arthritis, infectious arthritis, traumatic arthritis, carpal tunnel syndrome, bursitis or synovitis.
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| Publication number | Priority date | Publication date | Assignee | Title |
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1679622A (en) * | 2002-12-30 | 2005-10-12 | 胡锦心 | Use of sodium selenite in preparing medicine for osseous arthritis |
| WO2009043106A1 (en) * | 2007-10-03 | 2009-04-09 | Velacor Therapeutics Pty Ltd | Inorganic selenium and angiogenesis |
| CN103040864A (en) * | 2012-12-25 | 2013-04-17 | 深圳福山生物科技有限公司 | Medical application of selenium phosphate compound |
| CN104119237A (en) * | 2014-04-23 | 2014-10-29 | 深圳福山生物科技有限公司 | Choline salts of selenium-containing compounds, preparation method and applications thereof |
| CN104817608A (en) * | 2015-04-15 | 2015-08-05 | 深圳福山生物科技有限公司 | Cordycepin salt containing selenium compound, preparation method of cordycepin salt and application |
| CN108042564A (en) * | 2017-12-15 | 2018-05-18 | 合肥中龙神力动物药业有限公司 | A kind of sodium selenite vitamin C parenteral solutions and its manufacturing method |
| CN109512836A (en) * | 2018-12-21 | 2019-03-26 | 江西润泽药业有限公司 | A kind of sodium selenite vitamin C injection and preparation method thereof |
| CN110420185A (en) * | 2019-07-10 | 2019-11-08 | 西安交通大学 | A kind of preparation process of chondroitin sulfate A (CSA) nanometer selenium freeze drying powder injection |
-
2019
- 2019-12-31 CN CN201911420502.9A patent/CN110917209A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1679622A (en) * | 2002-12-30 | 2005-10-12 | 胡锦心 | Use of sodium selenite in preparing medicine for osseous arthritis |
| WO2009043106A1 (en) * | 2007-10-03 | 2009-04-09 | Velacor Therapeutics Pty Ltd | Inorganic selenium and angiogenesis |
| CN103040864A (en) * | 2012-12-25 | 2013-04-17 | 深圳福山生物科技有限公司 | Medical application of selenium phosphate compound |
| CN104119237A (en) * | 2014-04-23 | 2014-10-29 | 深圳福山生物科技有限公司 | Choline salts of selenium-containing compounds, preparation method and applications thereof |
| CN104817608A (en) * | 2015-04-15 | 2015-08-05 | 深圳福山生物科技有限公司 | Cordycepin salt containing selenium compound, preparation method of cordycepin salt and application |
| CN108042564A (en) * | 2017-12-15 | 2018-05-18 | 合肥中龙神力动物药业有限公司 | A kind of sodium selenite vitamin C parenteral solutions and its manufacturing method |
| CN109512836A (en) * | 2018-12-21 | 2019-03-26 | 江西润泽药业有限公司 | A kind of sodium selenite vitamin C injection and preparation method thereof |
| CN110420185A (en) * | 2019-07-10 | 2019-11-08 | 西安交通大学 | A kind of preparation process of chondroitin sulfate A (CSA) nanometer selenium freeze drying powder injection |
Non-Patent Citations (3)
| Title |
|---|
| 于庆臣等: "亚硒酸钠治疗牛前胃弛缓和瘤胃积食", 《畜牧兽医科技信息》 * |
| 杨玉东主编: "《生物医学纳米磁性材料原理及应用》", 31 October 2005, 吉林人民出版社 * |
| 沈霞等主编: "《现代生物化学检验与临床实践》", 31 October 1999, 上海科学技术文献出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022251810A1 (en) * | 2021-05-25 | 2022-12-01 | Baxter International Inc. | Containers with selective dissolved gas content |
| US11944586B2 (en) | 2021-05-25 | 2024-04-02 | Baxter International Inc. | Containers with selective dissolved gas content |
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