阅读说明：本技术 治疗椎间盘退行性疾病的组合物、脂质体纳米给药系统及其制备方法和应用 (Composition for treating intervertebral disc degenerative disease, liposome nano drug delivery system, preparation method and application thereof ) 是由 查建林 李晨光 于 2019-04-10 设计创作，主要内容包括：本发明公开了一种治疗椎间盘退行性疾病的组合物、脂质体纳米给药系统及其制备方法和应用,所述组合物是由齐墩果酸和淫羊藿苷按质量比为1:10～10:1组成。所述脂质体纳米给药系统,是由组合物、表面活性剂TPGS、胆固醇和磷脂制成的纳米脂质体制剂。本发明提供的由齐墩果酸和淫羊藿苷组成的组合物具有益气化瘀补肾的功效,可用于制备治疗椎间盘退行性疾病的药物；尤其是本发明提供的组合物的脂质体纳米给药系统,可有效提高组合物中齐墩果酸和淫羊藿苷的生物利用度,降低组合物的服用剂量和毒性,服用方便、量小高效且具有一定被动靶向性,提高了患者的顺应性,具有推广应用价值。(The invention discloses a composition for treating intervertebral disc degenerative diseases, a liposome nano drug delivery system, a preparation method and application thereof, wherein the composition is composed of oleanolic acid and icariin in a mass ratio of 1: 10-10: 1. The liposome nano drug delivery system is a nano liposome preparation prepared from a composition, a surfactant TPGS, cholesterol and phospholipid. The composition consisting of the oleanolic acid and the icariin has the effects of tonifying qi, removing blood stasis and tonifying the kidney, and can be used for preparing a medicament for treating the intervertebral disc degenerative diseases; particularly, the liposome nano drug delivery system of the composition provided by the invention can effectively improve the bioavailability of oleanolic acid and icariin in the composition, reduce the administration dosage and toxicity of the composition, is convenient to take, small in dosage, high-efficiency and has certain passive targeting property, the compliance of patients is improved, and the liposome nano drug delivery system has popularization and application values.)

1. A composition for treating a degenerative disc disease, comprising: the medicine is prepared from oleanolic acid and icariin in a mass ratio of 1: 10-10: 1.

2. Use of a composition according to claim 1, wherein: is used for preparing the medicine for treating the degenerative disc disease.

3. A liposomal nanodrug delivery system comprising the composition of claim 1, wherein: is a nanoliposome preparation prepared from the composition of claim 1, surfactant TPGS, cholesterol and phospholipid.

4. The liposomal nanodrug delivery system of claim 3, wherein: the particle size of the nano drug delivery system is 80-200 nm, the zeta potential is 0 +/-30 mV, the encapsulation rate is 75-99%, and the drug loading rate is 3-15%.

5. The liposomal nanodrug delivery system of claim 3, wherein: the mass ratio of the composition to the phospholipid is 1: 5-1: 30, the mass ratio of the surfactant TPGS to the phospholipid is 1: 8-1: 50, and the mass ratio of the cholesterol to the phospholipid is 1: 1-1: 20.

6. The liposomal nanodrug delivery system of claim 3 or 5, wherein: the phospholipid is one or more of soybean phospholipid, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoylphosphatidylethanolamine, vitamin E succinate polyethylene glycol ester, dilauroylphosphatidylcholine, dioleoylphosphatidylcholine, dilauroylphosphatidylglycerol, and dipalmitoylphosphatidylethanolamine.

7. A method of preparing the liposomal nanodrug delivery system of claim 3, wherein: the liposome nano drug delivery system is prepared by mixing the composition, a surfactant TPGS, cholesterol and phospholipid and then adopting a film dispersion method.

8. The method of claim 7, wherein:

dissolving oleanolic acid, surfactant TPGS, cholesterol and phospholipid in the composition in an organic solvent, performing reduced pressure rotary evaporation, and removing the organic reagent to obtain an organic phase; dissolving icariin in the composition in a hydration medium, adding an organic phase, stirring in a water bath for 30 minutes to 1 hour, performing ultrasonic treatment by an ultrasonic cell crusher for 2 minutes to 6 minutes, and performing membrane filtration to obtain the liposome nano drug delivery system.

9. The method of claim 8, wherein: the hydration medium is phosphate buffer salt solution with the pH value of 5-8 and the concentration of 1-20 mmol/ml.

10. Use of the liposomal nanodrug delivery system of claim 3, wherein: is used for preparing the medicine for treating the degenerative disc disease.

Technical Field

The invention relates to a composition for treating intervertebral disc degenerative diseases, a liposome nano drug delivery system, a preparation method and application thereof, and belongs to the technical field of medicines.

Background

Intervertebral disc degeneration (IVDD) is an aging disease characterized typically by the loss of extracellular matrix. According to the beginning factor, the intermediate link and the ending of intervertebral disc degeneration published by researchers such as the Wang champion in the Chinese medicine J. orthopedics and traumatology in the 3 rd year 2000, the nutrition provided by the cartilage endplate passage and the fiber ring peripheral passage is limited under the influence of the congenital structure of poor blood supply. IVDD is further exacerbated by aging, cell senescence, altered gene phenotype, increased metabolic capacity, and extracellular matrix disorders. Diseases such as intervertebral disc protrusion caused by IVDD are closely related to lumbosacral pain, and have great influence on human body, mind, society and economy.

The methods of traditional Chinese medicine are different when diagnosing and treating degenerative diseases of lumbar intervertebral discs, but most of them are good at tonifying qi, promoting qi, activating blood circulation and tonifying kidney. The professor of the university of Shanghai medicine, namely the professor of the Chinese wolfberry follows the theory of traditional Chinese medicine syndrome differentiation and treatment of 'taking qi as the main part, taking blood as the first part, taking phlegm and blood stasis into consideration, and treating liver, spleen and kidney simultaneously', and forms a qi-tonifying, blood stasis-removing and kidney-tonifying prescription for treating the degenerative diseases of the intervertebral disc through long-term clinical practice, and the clinical curative effect shows that the prescription has particularly remarkable curative effect on the protrusion of the lumbar intervertebral disc. However, the formula belongs to the traditional Chinese medicine formula, and after a doctor prescribes the formula, patients need to take the medicinal materials home to decoct the medicinal materials for taking, so that the formula is time-consuming, labor-consuming and inconvenient to take, and is difficult to adapt to the fast-paced life style of modern people. Therefore, a modern Chinese medicinal preparation form with convenient taking, small amount and high efficiency is urgently needed.

Oleanolic acid is mainly extracted from leaves of Olea europaea L of Oleaceae; fruit of Ligustrum lucidum ait; whole plant of swertia mileensis t.n.he etw.l.shi of Gentianaceae; mussotii franch, a mussotii; leaf and root of Apium graveolens Maxtia major of Umbelliferae; root bark and stem bark of Aralia chinensis L. of Araliaceae; root tuber of Cucurbitaceae plant Hemsleya macrocarpa c.y.wu, Hemsleya amabilis diels, Hemsleya amabilis cog. The oleanolic acid is clinically used for treating infectious acute icteric hepatitis, has obvious glutamic-pyruvic transaminase reducing and jaundice treating effects, and improves symptoms, physical signs and liver functions of patients with viral and chronic persistent hepatitis; it can also be used for treating psoriasis, rheumatic arthritis, nephritic edema, ascites due to liver cirrhosis, acute and chronic hepatitis, gastralgia, stranguria with turbid urine, metrorrhagia, traumatic injury, carbuncle, swelling, soreness of waist and knees, and threatened abortion. Chinese patent CN201510950032.2 proposes that oleanolic acid can be used as an alpha 36 estrogen receptor activator for preparing medicines for treating degenerative disc diseases.

Icariin is dry stem and leaf extract of Epimedium sagittatum Maxim, Epimedium pubescens Maxim, Epimedium wushanense Wu, and Epimedium koreanum nakai etc., and has effects of increasing cardiovascular and cerebrovascular blood flow, promoting hemopoietic function, immunity function and bone metabolism, invigorating kidney, tonifying yang, and resisting aging etc.

Because oleanolic acid is a fat-soluble component and icariin is a water-soluble component, a proper solvent is difficult to find and simultaneously the solubility requirements of the oleanolic acid and the icariin are met during injection administration; when the compound is prepared into solid preparations such as tablets, capsules and the like, the problems of low bioavailability and large administration dosage can occur during oral administration; therefore, at present, no report about the application of the composition of oleanolic acid and icariin in treating the intervertebral disc degenerative diseases exists.

Disclosure of Invention

The invention aims to provide a composition for treating intervertebral disc degenerative diseases, a liposome nano drug delivery system, a preparation method and application thereof.

A composition for treating intervertebral disc degenerative diseases comprises oleanolic acid and icariin in a mass ratio of 1: 10-10: 1.

The composition is used as an active ingredient to prepare a medicament for treating the intervertebral disc degenerative disease.

A liposome nanometer administration system of composition for treating intervertebral disc degenerative disease is a nanometer liposome preparation prepared from the composition, surfactant TPGS, cholesterol and phospholipid.

Furthermore, the particle size of the nano drug delivery system is 80-200 nm, the zeta potential is 0 +/-30 mV, the encapsulation rate is 75-99%, and the drug loading rate is 3-15%.

Furthermore, in the liposome nano drug delivery system, the mass ratio of the composition to the phospholipid is 1: 5-1: 30, the mass ratio of the surfactant TPGS to the phospholipid is 1: 8-1: 50, and the mass ratio of the cholesterol to the phospholipid is 1: 1-1: 20.

Further, the phospholipid is one or more of Soybean Phospholipid (SPC), Dipalmitoylphosphatidylcholine (DPPC), Distearoylphosphatidylcholine (DSPC), Dimyristoylphosphatidylcholine (DMPC), distearoylphosphatidylethanolamine (DSPE-PEG2000), vitamin E succinate polyethylene glycol ester (TPGS), Dilauroylphosphatidylcholine (DLPC), Dioleoylphosphatidylcholine (DOPC), Dilauroylphosphatidylglycerol (DOPG), dipalmitoylphosphatidic acid (DPPA) and Dipalmitoylphosphatidylethanolamine (DPPE).

The preparation method of the liposome nano drug delivery system comprises the steps of mixing the composition, the surfactant TPGS, cholesterol and phospholipid, and then preparing the liposome nano drug delivery system by adopting a film dispersion method.

Further, the preparation of the liposome nano drug delivery system comprises the following operations:

dissolving oleanolic acid, surfactant TPGS, cholesterol and phospholipid in the composition in an organic solvent, performing reduced pressure rotary evaporation, and removing the organic reagent to obtain an organic phase; dissolving icariin in the composition in a hydration medium, adding an organic phase, stirring in a water bath for 30 minutes to 1 hour, performing ultrasonic treatment by an ultrasonic cell crusher for 2 minutes to 6 minutes, and performing membrane filtration to obtain the liposome nano drug delivery system.

Further, the hydration medium is phosphate buffered saline (PBS buffer) with the pH value of 5-8 and the concentration of 1-20 mmol/ml.

Furthermore, the dosage form of the nano drug delivery system is injection or freeze-dried powder injection.

Furthermore, the freeze-drying protective agent used in the freeze-dried powder injection is one or more of lactose, sucrose, glucose, trehalose, mannitol and sorbitol, and the dosage of the freeze-dried protective agent accounts for 2-10% of the mass of the nano drug delivery system.

The application of the liposome nano drug delivery system is used for preparing the drug for treating the intervertebral disc degenerative disease.

Compared with the prior art, the invention has the following remarkable beneficial effects:

the composition consisting of the oleanolic acid and the icariin has the effects of tonifying qi, removing blood stasis and tonifying the kidney, and can be used for preparing a medicament for treating the intervertebral disc degenerative diseases; particularly, the liposome nano-drug delivery system of the composition provided by the invention can encapsulate the composition in a lipid bilayer to form a micro-vesicular body, wherein the bilayer structure has a lipophilic end and a hydrophilic end, can encapsulate fat-soluble and water-soluble components at the same time, has the same structure with a skin cell membrane, is easy to fuse with the cell membrane, can improve the bioavailability of the drug, is phagocytized by macrophages when the liposome enters the body, has a certain natural targeting effect, can further improve the bioavailability of oleanolic acid and icariin in, reduces the administration dosage and toxicity of the composition, is convenient to take, small in dosage, high in efficiency, has a certain passive targeting property, improves the compliance of patients, and has popularization and application values.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.