CN101502678B - Mixed gel for injection and preparation method thereof - Google Patents

Abstract

The invention relates to a mixed gel applied to injection and a preparation method thereof. The mixed gel of the invention is characterized by comprising polyaspartic acid terpenoid gel particle, cross bonding hyaluronic acid gel particle and hyaluronic acid or salt solution thereof, wherein, the volume of the polyaspartic acid terpenoid gel particle accounts for 5-80%, the volume ratio of the cross bonding hyaluronic acid gel particle and hyaluronic acid or salt solution thereof ranges from 1:19-19:1; the preparation method of the mixed gel has the following steps: respectively preparing the polyaspartic acid terpenoid gel particle and cross bonding hyaluronic acid gel particle; mixing evenly the particles with hyaluronic acid dissolved in the isosmotic solution after full swelling and balancing in isosmotic solution, sterilizing and packaging the mixture to obtain the finished product. The mixed gel of the invention can be used to prepare injection for beauty care and medical treatment and has the characteristics of long hold time and little side effect.

Description

A kind of mixed gel for injection and preparation method thereof

technical field

The invention belongs to the field of medicine, and relates to a mixed gel for injection and a preparation method thereof. The mixed gel is composed of polyaspartic acid compound gel particles, cross-linked hyaluronic acid gel particles and hyaluronic acid or its salt solution, and can be used for skin soft tissue and intra-articular injection.

Background technique

There are many types of polyaspartic acid compounds, and the current research and application are more widely used polyaspartic acid (polyaspartic acid, PASP) or its salt, polyaspartamide (polyaspartamide, PASPA) or its derivatives, polyaspartic acid Copolymers, etc., are a new generation of degradable biopolymer materials. Natural PASP exists in the shells of mollusks and snails, and the relative molecular mass ranges from 1000 to hundreds of thousands. PASP is a polycarboxylic acid with amino groups. In addition to the characteristics of general polycarboxylic acid, it also has good biocompatibility and biodegradability, and is non-toxic and pollution-free. It has a wide range of uses. Therefore, since its first artificial Since its synthesis, its research has been carried out in depth all over the world. At present, many companies at home and abroad have achieved industrialization, and the products are mainly used in sewage treatment, daily chemical industry, oilfield chemical industry, pesticide, fertilizer and pharmaceutical fields. With the deepening of research, PASPA and its derivatives, such as poly(hydroxypropyl)-DL-asparagine [α, β-poly(N-hydroxypropyl-DL-aspartamide), PHPA], poly(hydroxyethyl) -DL-asparagine [α, β-poly(N-hydroxyethyl-DL-aspartamide), PHEA], poly(propyl, 3-hydroxypropyl)-DL-aspartamide [Poly-(propyl, 3- Hydroxypropyl)-DL-asparamide, PHPPA] and aspartic hydrazide (α, β-polyasparthydrazide, PAHY), and polyaspartic acid copolymers have also been synthesized successively. A number of patents at home and abroad have disclosed the preparation process of PASP and PASPA and their derivatives (such as CN1123588, CN1195000, CN1814650, CN101003626, etc.), and the application research in the field of medicine mainly focuses on drug sustained release and tissue engineering materials. PASP and PHEA can be used as plasma expanders (Antoni G, Neri P. Biopolymers. 1974, 13: 1721-172; Giammona G, Carlisi B, Cavallaro G, et al. J Controlled Release, 1994, 29: 63-72. ), PHPPA, PHEA derivatives and glutaraldehyde cross-linked PAHY hydrogel are good drug sustained-release carriers (Giammona G, Pitarresi G, Tomarchio V, JContro Rel, 1996, 41:195; Liu Zhenhua, Wang Jian, Fan Changlie , et al. Acta Polymer Sinica, 1998, 5: 562; Zhou Tao, Wang Bin, Tang Guping, etc. Biomedical Engineering Journal, 1999, 16(4): 423-428), PHEA chitosan membrane can be used as a substrate for cell growth Scaffold (Zhu Aiping, Wang Shiquan, Cheng Daming, etc. Acta Bioengineering, 2002, 18:109), PHEA hydrogel prepared by photocrosslinking becomes a less toxic biomaterial due to less impurities (Gammona G, PitarresiG , Cavallaro G, et al. Biochemica et Biophysica Acta, 1999, 1428: 29).

Polyaspartic acid compounds are a kind of artificial chemical synthetic materials. Compared with animal-derived collagen materials, they reduce the risk of allergic reactions and transmission of infectious diseases, and are easy to be accepted by more people. At the same time, polyaspartic acid compounds have an amide bond structure similar to proteins, and are biodegradable materials. The degradation products are small molecules of amino acids, which can eventually be degraded into water and carbon dioxide, and have good biocompatibility. Existing data prove that PASP is non-toxic and non-mutagenic [Yang Shilin et al., Journal of Environment and Health. 2004, 11, 21(6): 398～401], the subcutaneous implantation test of PHEA, PHPA and PHPPA materials in mice proves that they can be gradually absorbed in the body, and have no obvious toxic effects on liver and kidney tissues, hemoglobin, white blood cells, etc. It is a kind of polymer material with good tissue compatibility [Weng Lihong et al. Science and Technology Bulletin, 1999, 15(3) 161].

Polyaspartic acid compounds are non-toxic, good biocompatibility, and need a period of time to degrade and disappear in the body, making them suitable for tissue filling materials; after absorbing water, they form a hydrogel with certain strength and elasticity. It also has the conditions to become a joint cavity lubricant and a liner. But there is no relevant research and report yet.

)，现已广泛用作抗粘连性手术植入物、骨关节病治疗首选生物医学材料、药物缓释基质、软组织增大替代物、美容整形材料等；缩水甘油醚交联的HA凝胶Restylan

和Perlane

系列产品(由瑞士Q-Med AB公司生产)应用于隆唇、改善面部皱纹和褶皱，效果可维持到半年以上。Hyaluronic acid (HA) is an important component of skin, vitreous, synovial fluid and cartilage tissue, and has good lubricity, viscoelasticity and non-immunogenicity (Laurent TC et al., Ann.Rheμm.Dis.1995 , 54: 429-432). Now a large number of HA clinical products are used as viscoelastic agents in ophthalmology, viscoelastic supplements in orthopedics, and preventive agents for postoperative adhesions (Ling Peixue and He Yanli, etc., Chinese Journal of Biochemical Medicine, 1998, 19: 200-204), Polymer sodium hyaluronate (SH) solutions are also used to inject facial skin to reduce wrinkles, improve skin appearance and structure (RU2272634, RU2194512) or improve lip shape and prevent skin aging (RU2159111). However, natural HA is easily degraded by enzymes in body tissues, and its good water solubility makes it easy to diffuse in tissues, so that its local retention time in animal tissues is very short. Cross-linked HA can overcome these defects while retaining its good biocompatibility and non-immunogenicity, so it is currently an ideal tissue filling material. Such as divinyl sulfone cross-linked HA gel hylan series products (such as Hylaform of Allergan of the United States , Biomatrix Synvisc

), has been widely used as anti-adhesion surgical implants, biomedical materials of choice for osteoarthritis treatment, drug slow-release matrix, soft tissue augmentation substitutes, cosmetic plastic materials, etc.; Glycidyl ether cross-linked HA gel Restylan

and Perlane

A series of products (manufactured by Q-Med AB, Switzerland) are used for lip augmentation and improvement of facial wrinkles and folds, and the effect can be maintained for more than half a year.

The invention adopts the combination of cross-linked HA and polyaspartic acid compounds, and simultaneously adds uncross-linked HA and its salt solution as a carrier. Compared with collagen or other chemical synthetic materials, the product of the present invention has good biocompatibility, less side effects, and is easy to be accepted by patients; The presence of particles can increase the retention time of the product in the body tissue and keep the product in the body tissue. HA and its salt solution provide a lubricated and elastic space, which makes the suspended particles with a certain strength easier to inject; the elastic cross-linked HA particles and the HA and its salt solution as a carrier can make the filling site The appearance is more soft and natural, which improves the plasticity of the product.

The invention can be used for local injection to treat arthritis. The solution of HA and its salt as carrier can immediately enhance the lubricating effect of joint synovial fluid, and polyaspartic acid compound gel particles and cross-linked HA gel particles can enhance joint synovial fluid With the cushioning effect of external force, the mixture of the three becomes an effective isolation and lubricating pad, which can exist in the joint cavity for a long time, so that the damaged cartilage can be fully repaired and the root cause of inflammation can be eliminated.

In summary, the hybrid gel of the present invention has the characteristics of easy injection, long local retention time, good plasticity, and few side effects.

Contents of the invention

The object of the present invention is to provide a mixed gel and its preparation method, and use the mixed gel for the preparation of cosmetic or medical injections, such as the preparation of intradermal or subcutaneous injections for eliminating facial wrinkles, or for the treatment of Intra-articular injection for arthritis.

The hybrid gel of the present invention is composed of polyaspartic acid compound gel particles, cross-linked HA gel particles and a solution of HA and its salt.

The volume of polyaspartate compound gel particles in the mixed gel of the present invention accounts for 5%-80%.

The average particle diameter of polyaspartic acid compound gel particles in the mixed gel of the present invention is in the range of 1-5000 μm, preferably 100-1000 μm, more preferably 250-500 μm. Specifically, observe under a microscope and measure the diameter of the PASP particles, wherein more than 50% of the particles have a diameter ranging from 1 to 5000 μm, preferably more than 50% of the particles have a diameter ranging from 100 to 1000 μm, more preferably more than 50% The particle diameter ranges from 250 to 500 μm. Since the polyaspartic acid compound gel particles are translucent, colorless or light yellow in solution, the appearance of the hybrid gel of the present invention is a colorless or light yellow viscous body with opalescence.

The polyaspartic acid compound in the hybrid gel of the present invention is selected from, but not limited to, PASP or its salt, PASPA or its derivative, and one, any two or more than two kinds of polyaspartic acid copolymers Mixtures, preferably polyaspartic acid and/or polyasparagine, more preferably polyaspartic acid. Salts of PASP such as sodium polyaspartate (PASP-Na), PASPA or its derivatives such as PHPA, PHEA, PHPPA, PAHY, glutaraldehyde-crosslinked PASP, PHEA or PAHY products, photocrosslinked PHEA or PAHY products etc., polyaspartic acid copolymers such as PASP and polylactic acid (polylactic acid) block copolymer (PASP-PLA), PASP and polyethylene glycol (PEG) block copolymer (PASP-PEG), L- Aspartic acid (L-aspartic acid, L-Asp) and glycine (Glycine) copolymer (PASP-PGLY), L-Asp and 6-aminohexanoic acid (aminohexanoic acid) copolymer (PASP-PAHA), L - Copolymer of Asp and glutamic acid (PASP-PGLU), copolymer of L-Asp and lysine (PASP-PLYS), etc.

The volume ratio of the cross-linked HA gel particles in the hybrid gel of the present invention to HA and its salt solution ranges from 1:19 to 19:1, preferably 4:1 to 1:4, more preferably 3:1 to 1:3 . Since the cross-linked HA gel particle is a water-insoluble water-containing particle fully swollen in the solution, it will not re-swell or shrink in the HA solution, so the volume of the cross-linked HA gel particle and HA and its salt solution Ratio limits the proportioning range of the two to 1:19 to 19:1, preferably 4:1 to 1:4, more preferably 3:1 to 1:3.

The cross-linked HA in the hybrid gel of the present invention is selected from divinyl sulfone cross-linked HA products or glycidyl ether cross-linked HA products or other cross-linked HA products, preferably glycidyl ether cross-linked HA products, more preferably using 1,4-Butanediol diglycidyl ether (BDDE) cross-linked HA product.

HA or its salt in the mixed gel of the present invention is selected from SH, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, ammonium hyaluronate, tetrabutylammonium hyaluronate, bismuth hyaluronate, hyaluronic acid Zinc acid or one of other hyaluronates, or a mixture of any two or more of them, preferably SH.

The mixed gel of the present invention can be used to prepare injections for cosmetic or medical purposes, for example, to prepare cosmetic injections for improving facial wrinkles and reducing skin folds, for local subcutaneous or intradermal injection, and for preparation of joint cavity injections for treating arthritis.

Mixed gel of the present invention can be prepared as follows:

① Prepare polyaspartic acid compound gel particles, fully swell and balance in isotonic solution, and obtain water-insoluble hydrogel particles;

② Prepare cross-linked HA particles, fully swell and balance in isotonic solution to obtain water-insoluble hydrogel particles;

③ Dissolving HA or its salt in an isotonic solution to obtain an isotonic solution of HA or its salt;

④ Mix the hydrogel particles obtained in ① and ② with the isotonic solution of HA or its salt obtained in ③ evenly, sterilize and pack separately to obtain the finished product.

The isotonic solution described in the preparation method of the present invention is a sodium chloride solution or a phosphate buffer solution with an osmotic pressure ranging from 250 to 350 mOsmol/L, preferably a phosphate buffer solution with an osmotic pressure of 250 to 350 mOsmol/L and a pH of 6.5 to 7.5. For example, 0.9% sodium chloride solution (physiological saline), or a buffer solution containing 0.3 mM sodium dihydrogen phosphate, 1.6 mM disodium hydrogen sulfate, 146.5 mM sodium chloride, and adjusting pH 6.5 to 7.5 with hydrochloric acid.

The polyaspartic acid compound described in step ① of the preparation method of the present invention is selected from but not limited to PASP or its salt, PASPA or its derivatives, one, any two, or A mixture of two or more, preferably PASP and/or PAHY, more preferably PASP. Salts of PASP such as sodium polyaspartate (PASP-Na), PASPA or its derivatives such as PHPA, PHEA, PHPPA, PAHY, glutaraldehyde-crosslinked PASP, PHEA or PAHY products, photocrosslinked PHEA or PAHY products etc. Polyaspartic acid copolymers such as PASP-PLA, PASP-PEG, PASP-PGLY, PASP-PAHA, PASP-PGLU, PASP-PLYS, etc.

The average particle diameter of the polyaspartate compound gel particles described in step ① of the preparation method of the present invention ranges from 1 to 5000 μm, preferably 100 to 1000 μm, more preferably 250 to 500 μm.

The cross-linked HA gel described in step ② of the preparation method of the present invention is selected from divinyl sulfone cross-linked HA products, and can also be HA products cross-linked with glycidyl ether and other cross-linked HA products, preferably using Glycidyl ether cross-linked HA products, more preferably BDDE cross-linked HA products.

The HA and its salts described in step ③ of the preparation method of the present invention are selected from SH, potassium hyaluronate, calcium hyaluronate, magnesium hyaluronate, ammonium hyaluronate, tetrabutylammonium hyaluronate, hyaluronic acid One of bismuth and zinc hyaluronate or other hyaluronates, or a mixture of any two or more of them, preferably SH.

The preparation method of the invention has simple process and easy control of product quality; the prepared mixed gel of PASP and cross-linked HA is aseptic and pyrogen-free, and the product is uniform and delicate, and is easy to inject.

The mixed gel of the invention is suitable for preparing injections for cosmetic and medical purposes, and has the advantages of easy injection, long local retention time, good plasticity, less side effects and the like. It is used to prepare fillers for injection into facial skin, and has the advantages of good wrinkle elimination effect and long maintenance time. It is used to prepare the joint cavity injection for the treatment of arthritis. It can be used as an isolation and lubricating pad in the joint cavity for a long time. It has sufficient buffering effect on the external force, so that the cartilage tissue can be fully repaired. It has the advantages of less injection times, local It has the characteristics of less stimulation and good curative effect.

Detailed ways

The following examples are to better illustrate the present invention, not to limit the present invention.

Embodiment one

References [Fang Li and Tan Tianwei, Chemical Reaction Engineering and Technology, 2003, 19 (4): 295-299] method to prepare PASP: take L-ASP as raw material, phosphoric acid as catalyst, sulfolane as solvent, and heat to 200 °C, reacted for 4.5h, and removed the generated water during the reaction. After the reaction is completed, the reaction product is repeatedly washed with methanol and water until neutral, and dried under reduced pressure at 40° C. to obtain a reaction intermediate polysuccinimide (PSI) with a molecular weight of about 400,000 to 500,000. Under the condition of ice bath, take PSI and add 5-6 times of 2mol/L NaOH solution to hydrolyze for 15min, then dropwise add methanol-NaCl saturated solution to form a precipitate, leave it overnight, filter, collect the precipitate and dry it under reduced pressure at 40°C to obtain PASP solid. Disperse PASP in deionized water, pass through 80-mesh sieve, add an equal volume of phosphate buffered saline (PBS) with 2 times isotonic concentration (each 1000ml contains NaH ₂ PO ₄ 2H ₂ O 90mg, Na ₂ HPO ₄ 12H ₂ O 1.12g, NaCl 17g), incubated at 65°C for 4h, centrifuged to remove the supernatant, and precipitated with isotonic PBS (each 1000ml contains NaH ₂ .PO ₄ ·2H ₂ O 45mg, Na ₂ HPO ₄ ·12H ₂ O 0.56g , NaCl 8.5g, pH 7.2) washed several times, and centrifuged to remove the supernatant to obtain PASP gel particles with an average particle size ranging from about 200 to 250 μm.

Dissolve 10g SH (average molecular weight 1.2 million Daltons) in 80ml 1% NaOH solution containing 0.5g BDDE. React at 50°C for 4 hours, purify with deionized water at 70°C for 10 hours, squeeze the gel through a 60-mesh sieve, collect the gel particles, add an equal volume of PBS with 2 times the isotonic concentration, and equilibrate for 4 hours to obtain an average particle size of about HA gel particles of 180 μm.

Another SH (average molecular weight 1 million Daltons) was dissolved in isotonic PBS to prepare an isotonic SH solution with a concentration of 1%.

Take 5ml of PASP gel particles prepared above and 75ml of cross-linked HA gel particles and mix them with 20ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix Gel 1A.

Take 10ml of PASP gel particles prepared above and 70ml of cross-linked HA gel particles and mix them with 20ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 1B.

Take 25ml of PASP gel particles prepared above and 65ml of cross-linked HA gel particles and mix them with 10ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 1C.

Take 40ml of PASP gel particles prepared above and 40ml of cross-linked HA gel particles and mix them with 20ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix Gel 1D.

Take 50ml of PASP gel particles prepared above and 30ml of cross-linked HA gel particles and mix them with 20ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 1E.

Take 70ml of PASP gel particles prepared above, 20ml of cross-linked HA gel particles and 10ml of SH solution, mix them with 100°C circulating steam twice, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 1F.

Take 75ml of PASP gel particles prepared above, 10ml of cross-linked HA gel particles and 15ml of SH solution, mix them with 100°C flowing steam twice, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 1G.

Take 80ml of PASP gel particles prepared above and 15ml of cross-linked HA gel particles and mix them with 5ml of SH solution, sterilize twice with circulating steam at 100°C for 30 minutes each time, and aseptically dispense them into disposable syringes to obtain the mixture of the present invention. Gel 1H.

Embodiment two

Reference method (CN1814650) to prepare PASP resin: prepare PSI according to the method described in Example 1, crush PSI, pass through a 160-mesh sieve, add 30 times deionized water, continue stirring for 8-10 hours, and add cross-linking agent hexamethylenediamine at intervals When the concentration reaches 0.5%, add 3 to 4 times of ethanol after the stirring is completed, let it stand for 3 days, collect the precipitate and dry it under reduced pressure at 25°C, crush the dried product, pass through a 160-mesh sieve, add 20 times of 50% ethanol solution, and stir for 30 minutes. Add 20% NaOH solution dropwise to the reaction solution until the reaction liquid is in a colloidal state, add 3 to 5 times of ethanol, place it for 3 days, collect the precipitate, wash and purify it with ethanol and deionized water alternately for several times, finally collect the precipitate from ethanol, and reduce it at 40°C Press and dry to obtain PASP resin. Disperse the PASP resin in deionized water, pass through a 40-mesh sieve, add an equal volume of PBS with 2 times the isotonic concentration, incubate at 65°C for 4 hours, remove the supernatant by centrifugation, wash the precipitate several times with isotonic PBS, and remove the supernatant by centrifugation The solution obtained PASP gel particles with an average particle size ranging from 350 to 500 μm.

Mix 10g SH (average molecular weight 900,000 Daltons) with 60ml 1% NaOH solution containing 0.8g BDDE, react at 50°C for 4h, purify with deionized water at 70°C for 10h, squeeze the gel through a 40-mesh sieve , collect the gel particles, add an equal volume of 2 times isotonic PBS, and equilibrate for 4 hours to obtain cross-linked HA gel particles with an average particle size of about 500 μm.

Another SH (average molecular weight: 1.8 million Daltons) was dissolved in isotonic PBS to prepare an isotonic SH solution with a concentration of 1%.

Take 5ml of PASP gel particles prepared above and 70ml of cross-linked HA gel particles and mix them with 25ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 2A.

Take 10ml of PASP gel particles prepared above and 75ml of cross-linked HA gel particles and mix them with 15ml SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 2B.

Take 15ml of the PASP gel particles prepared above and 75ml of cross-linked HA gel particles and mix them with 10ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 2C.

Take 20ml of PASP gel particles prepared above and 60ml of cross-linked HA gel particles and mix them with 20ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Hybrid Gel 2D.

Take 25ml of PASP gel particles prepared above and 45ml of cross-linked HA gel particles and mix them with 30ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the mixture of the present invention. Gel 2E.

Take 30ml of PASP gel particles prepared above and 50ml of cross-linked HA gel particles and mix them with 20ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 2F.

Take 45ml of PASP gel particles prepared above and 45ml of cross-linked HA gel particles and mix them with 10ml SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 2G.

Take 60ml of PASP gel particles prepared above, 20ml of cross-linked HA gel particles and 20ml of SH solution, mix them with 20ml of SH solution, sterilize twice with circulating steam at 100°C, each time for 30min, and aseptically dispense them into disposable syringes to obtain the present invention. Mix gel 2h.

Take 80ml of PASP gel particles prepared above and 15ml of cross-linked HA gel particles and mix them with 5ml of SH solution, sterilize twice with circulating steam at 100°C for 30 minutes each time, and aseptically dispense them into disposable syringes to obtain the mixture of the present invention. Gel 2I.

Thrust test:

Under the pressure of 15～50N, the gel described in the embodiment of the present invention can be pushed out easily with No. 4～6 needles, and the gel described in the embodiment of the present invention can be easily pushed out with No. 5～7 needles. roll out.

Stability test:

The gel described in Examples 1 and 2 of the present invention was stored in a refrigerator at 4°C for one week, then taken out and kept at 35°C to 40°C for 10 minutes, the properties of the gel did not change, and it was in a uniform suspension state; Example 1 of the present invention 2. Put the gel described in 2 into a dialysis bag, and soak it in 37°C constant-temperature saline for 4 weeks, and the gel will lose no more than 20% of its weight.

Animal experiment:

1. Rat subcutaneous implantation test

Take the mixed gel 1D sample of Embodiment 1 of the present invention and inject it into the back of the rat subcutaneously. No reactions such as redness, swelling, and bruising were observed. After 4 weeks, the animal was killed, and the back skin was opened. It was observed that the implanted samples did not spread, Discoloration, thinner cysts formed around the implant, no adhesion to the surrounding tissue, no vascular proliferation. A mass is formed locally, and the surrounding tissue has no abnormal changes.

2. Local irritation test of rabbit joint cavity

A 2G sample of the mixed gel in Example 2 of the present invention was injected into the knee joint of the rabbit, each dose was 0.2ml/time, and the injection was repeated at intervals of one week, for a total of 2 injections. 48 hours after the second injection, samples were taken for histological examination, and the irritation of the samples to joint tissues was observed. No obvious redness and swelling were found after the samples were injected into the rabbit knee joints. Histological examination of cartilage and synovium showed that there was a small amount of inflammatory cell infiltration in the synovium of the joint, and no obvious abnormality was found in the articular cartilage tissue.

Claims (32)

1. a mixed gel for injection is characterised in that by poly-aspartate compounds gel particle, cross-linked hyaluronic acid gel granule and hyaluronic acid or its saline solution and forms; Wherein said poly-aspartate compounds is selected from a kind of, any two kinds or the two or more mixture in poly-aspartate or its salt, poly-asparagine or derivatives thereof, the poly-aspartate analog copolymer.

2. the described mixed gel of claim 1, its characteristic is that also wherein the volume of poly-aspartate compounds gel particle accounts for 5%～80%.

3. claim 1 or 2 arbitrary described mixed gels, the average particle size range of wherein said poly-aspartate compounds gel particle is 1～5000 μ m.

4. claim 1 or 2 arbitrary described mixed gels, the average particle size range of wherein said poly-aspartate compounds gel particle is 100～1000 μ m.

5. claim 1 or 2 arbitrary described mixed gels, the average particle size range of wherein said poly-aspartate compounds gel particle is 250～500 μ m.

6. the described mixed gel of claim 1, wherein said poly-aspartate compounds is selected from poly-aspartate and/or poly-asparagine.

7. the described mixed gel of claim 1, wherein said poly-aspartate compounds is a poly-aspartate.

8. the described mixed gel of claim 1, its characteristic is that also the volume ratio scope of said cross-linked hyaluronic acid gel granule and hyaluronic acid or its saline solution is 1: 19～19: 1.

9. the described mixed gel of claim 1, its characteristic is that also the volume ratio scope of said cross-linked hyaluronic acid gel granule and hyaluronic acid or its saline solution is 4: 1～1: 4.

10. the described mixed gel of claim 1, its characteristic is that also the volume ratio scope of said cross-linked hyaluronic acid gel granule and hyaluronic acid or its saline solution is 3: 1～1: 3.

11. claim 1,8,9 or 10 arbitrary described mixed gels, wherein said cross-linked-hyaluronic acid is selected from crosslinked hyaluronic acid product of divinylsulfone or the crosslinked hyaluronic acid product of glycidyl ether.

12. claim 1,8,9 or 10 arbitrary described mixed gels, wherein said cross-linked-hyaluronic acid is selected from the crosslinked hyaluronic acid product of glycidyl ether.

13. claim 1,8,9 or 10 arbitrary described mixed gels, wherein said cross-linked-hyaluronic acid is for adopting 1, the hyaluronic acid product that the 4-butanediol diglycidyl ether is crosslinked.

14. claim 1,8,9 or 10 arbitrary described mixed gels, wherein said hyaluronic acid or its salt are selected from a kind of, two or more mixture arbitrarily in hyaluronate sodium, potassium hyaluronate, calcium hyauronate, hyaluronic acid magnesium, hyaluronic acid ammonium, hyaluronic acid TBuA, bismuth hyalurate and the Curiosin.

15. claim 1,8,9 or 10 arbitrary described mixed gels, wherein said hyaluronic acid or its salt are hyaluronate sodium.

16. the described mixed gel of claim 1 is used to prepare the injection of beauty treatment or medical application.

17. a method for preparing the said mixed gel of claim 1 comprises the steps:

1.. preparation poly-aspartate compounds gel particle, abundant swelling, balance obtain water-insoluble aqueous gel granule in isosmotic solution;

2.. preparation cross-linked-hyaluronic acid granule, abundant swelling, balance obtain water-insoluble aqueous gel granule in isosmotic solution;

3.. hyaluronic acid or its salt are dissolved in the isosmotic solution, obtain the isosmotic solution of hyaluronic acid or its salt;

4.. will be 1. and 2. gained aqueous gel granule and the 3. hyaluronic acid of gained or the isosmotic solution mix homogeneously of its salt, sterilization back packing makes finished product.

18. the described method for preparing of claim 17, wherein said isosmotic solution are sodium chloride solution or the PBS of osmotic pressure scope at 250～350mOsmol/L.

19. the described method for preparing of claim 17, wherein said isosmotic solution are the PBS of osmotic pressure scope in 250～350mOsmol/L, pH6.5～7.5.

20. the described method for preparing of claim 17, wherein the poly-aspartate compounds of step described in 1. is selected from poly-aspartate and/or poly-asparagine.

21. the described method for preparing of claim 17, wherein the poly-aspartate compounds of step described in 1. is poly-aspartate.

22. the described method for preparing of claim 17, wherein the average particle size range of the poly-aspartate compounds gel particle of step described in 1. is 1～5000 μ m.

23. the described method for preparing of claim 17, wherein the average particle size range of the poly-aspartate compounds gel particle of step described in 1. is 100～1000 μ m.

24. the described method for preparing of claim 17, wherein the average particle size range of the poly-aspartate compounds gel particle of step described in 1. is 250～500 μ m.

25. the described method for preparing of claim 17, wherein the cross-linked-hyaluronic acid of step described in 2. is selected from crosslinked hyaluronic acid product of divinylsulfone or the crosslinked hyaluronic acid product of glycidyl ether.

26. the described method for preparing of claim 17, wherein the cross-linked-hyaluronic acid of step described in 2. is selected from the crosslinked hyaluronic acid product of glycidyl ether.

27. the described method for preparing of claim 17, wherein the cross-linked-hyaluronic acid of step described in 2. is for adopting 1, the hyaluronic acid product that the 4-butanediol diglycidyl ether is crosslinked.

28. the described method for preparing of claim 17, wherein the hyaluronic acid of step described in 3. or its salt are selected from a kind of, two or more the mixture arbitrarily in hyaluronate sodium, potassium hyaluronate, calcium hyauronate, hyaluronic acid magnesium, hyaluronic acid ammonium, hyaluronic acid TBuA, bismuth hyalurate and the Curiosin.

29. the described method for preparing of claim 17, wherein hyaluronic acid or its salt of step described in 3. is hyaluronate sodium.

30. the mixed gel that the arbitrary described method for preparing of claim 17～29 makes.

31. the described mixed gel of claim 30 is used to prepare the injection of beauty treatment or medical application.

32. the described mixed gel of claim 30 is used to prepare the joint cavity injection agent of treatment of arthritis.