技术领域technical field
本发明涉及一种包载重组人生长激素缓释药物微囊，属于药物技术领域。The invention relates to a slow-release medicine microcapsule loaded with recombinant human growth hormone, which belongs to the technical field of medicine.
技术背景technical background
1920年，人们发现了生长激素(human growth hormone, hGH )的存在，生长激素是腺垂体细胞分泌的蛋白质，是一种肽类激素，由191个氨基酸残基构成，相对分子质量为22124道尔顿。生长激素分子构造上具有4个α螺旋,使生长激素可以和其受体有良好结合。1958 年，人们发现可以将人生长激素用于治疗侏儒症，但是，由于当时仅仅能够从尸体的脑垂体中提取出人生长激素，数量有限，难以保证临床用量，而限制了其应用。1986年，波耶尔通过基因重组的技术成功的合成了重组人生长激素( recombinant human growth hormone, rhGH )，其结构与人垂体分泌的生长激素完全相同。这样便实现了重组人生长激素的大规模生产和应用。In 1920, people discovered the existence of growth hormone (human growth hormone, hGH). Growth hormone is a protein secreted by pituitary cells. It is a peptide hormone consisting of 191 amino acid residues and a relative molecular mass of 22124 dahl. pause. There are four α-helices in the molecular structure of growth hormone, so that growth hormone can be well combined with its receptor. In 1958, it was discovered that human growth hormone could be used to treat dwarfism. However, at that time, human growth hormone could only be extracted from the pituitary gland of the corpse, and the quantity was limited, so it was difficult to guarantee the clinical dosage, which limited its application. In 1986, Boyer successfully synthesized recombinant human growth hormone (rhGH) through gene recombination technology, and its structure is exactly the same as the growth hormone secreted by human pituitary gland. In this way, the large-scale production and application of recombinant human growth hormone has been realized.
正常情况下，生长激素呈脉冲式分泌，生长激素的分泌受下丘脑产生的生长激素释放素的调节，还受性别、年龄和昼夜节律的影响，睡眠状态下分泌明显增加。生长激素的主要生理功能是促进神经组织以外的所有其他组织生长、促进机体合成代谢和蛋白质合成、促进创面愈合等。在临床上，广泛应用重组人生长激素预防和治疗因生长激素缺乏导致的儿童生长发育迟缓、呆小症、先天性卵巢发育不全（即Turner综合症）、老年人免疫功能的低下、慢性肾脏疾病、手术或烧伤患者的创面愈合迟缓等病症。Under normal circumstances, growth hormone is secreted in pulses. The secretion of growth hormone is regulated by the growth hormone releasing hormone produced by the hypothalamus, and is also affected by gender, age and circadian rhythm. The secretion of growth hormone increases significantly during sleep. The main physiological function of growth hormone is to promote the growth of all other tissues except nerve tissue, promote the body's anabolism and protein synthesis, and promote wound healing. Clinically, recombinant human growth hormone is widely used to prevent and treat growth retardation, cretinism, congenital ovarian hypoplasia (Turner syndrome), low immune function of the elderly, and chronic kidney disease caused by growth hormone deficiency. , surgery or burn patients with delayed wound healing and other diseases.
虽然重组人生长激素临床应用广泛，但由于它是蛋白质类药物，在体内外环境可能经受多种复杂的化学降解和物理变化而失活，如凝聚、沉淀、消旋化、水解、脱酰氨基等。加之肾脏的排泄，使重组人生长激素在体内的半衰期短，因而治疗上述疾病一般都需要长期多次用药。目前，普遍采取一周三次或一天一次注射的方式给药。频繁注射给药容易导致患者的顺应性降低，同时加重了其经济负担。针对重组人生长激素用药周期长，且使用不方便的缺点，开发长效稳定的重组人生长激素是进一步拓宽生长激素应用范围的关键所在。目前所采取的方法主要是聚乙二醇化，即通过重组人生长激素上的氨基、羧基或巯基与端甲氧基聚乙二醇共价键合，有效阻止蛋白水解酶与重组人生长激素的直接接触，降低了其降解程度，进而降低了其清除率，改善了其稳定性，增加了其循环时间，减少了给药次数。Ross Clark等人于1996年最早研究了聚乙二醇偶联重组人生长激素的方法和活性（Journal of Biological Chemistry，1996，271,21969-21977）。Although recombinant human growth hormone is widely used clinically, because it is a protein drug, it may undergo a variety of complex chemical degradation and physical changes in vivo and in vitro, such as aggregation, precipitation, racemization, hydrolysis, and deamidation. wait. Combined with the excretion of the kidneys, the half-life of recombinant human growth hormone in the body is short, so the treatment of the above diseases generally requires long-term multiple medications. At present, it is generally administered three times a week or once a day. Frequent injections can easily lead to reduced compliance of patients and increase their economic burden. Aiming at the shortcoming of long-term medication cycle and inconvenient use of recombinant human growth hormone, the key to further broaden the application range of growth hormone is to develop long-acting and stable recombinant human growth hormone. The method currently adopted is mainly PEGylation, that is, through the covalent bonding of the amino, carboxyl or sulfhydryl groups on the recombinant human growth hormone with the terminal methoxy polyethylene glycol, effectively preventing the proteolytic enzyme from interacting with the recombinant human growth hormone. Direct contact reduces its degree of degradation, thereby reducing its clearance rate, improving its stability, increasing its circulation time, and reducing the number of administrations. Ross Clark et al first studied the method and activity of polyethylene glycol-conjugated recombinant human growth hormone in 1996 (Journal of Biological Chemistry, 1996, 271, 21969-21977).
王思勉在中国专利CN1253472C中，以一端为醛基的端甲氧基聚乙二醇与重组人生长激素中N端的α氨基共价键合，得到一种聚乙二醇重组人生长激素缀合物。吴自荣等人在中国专利CN1243022C中，将具有谷氨酰胺残基的重组人生长激素和一端氨基化的单甲氧基聚乙二醇，在转谷氨酰胺酶的催化下进行反应，在重组人生长激素谷氨酰胺残基上形成酰胺键，从而获得一种聚乙二醇修饰的重组人生长激素缀合物。CN101385858B，CN101491681B，CN1477126A，CN1565624A，CN10809038B等均采用相似的方法制备聚乙二醇化的重组人生长激素。但是，根据Ross Clark等人的研究结果，并非所有的聚乙二醇化的重组人生长激素都保持原有的活性，某些甚至起到副作用，以至于无法应用。In the Chinese patent CN1253472C, Wang Simian covalently bonded methoxypolyethylene glycol with one end as an aldehyde group to the α-amino group at the N-terminus of recombinant human growth hormone to obtain a recombinant human growth hormone conjugated with polyethylene glycol. compound. In the Chinese patent CN1243022C, Wu Zirong and others reacted recombinant human growth hormone with glutamine residues and monomethoxy polyethylene glycol aminated at one end under the catalysis of transglutaminase. An amide bond is formed on the glutamine residue of growth hormone to obtain a polyethylene glycol-modified recombinant human growth hormone conjugate. CN101385858B, CN101491681B, CN1477126A, CN1565624A, CN10809038B, etc. all adopt similar methods to prepare PEGylated recombinant human growth hormone. However, according to the research results of Ross Clark et al., not all PEGylated recombinant human growth hormones maintain their original activity, and some even have side effects, so that they cannot be used.
药物缓释制剂是另一种有效保护蛋白类药物在体内稳定长效作用的有效策略，主要包括脂质体（CN102188364A，CN101926770B），水凝胶（CN102989034A）和微囊（CN102302455B，CN102370630A）。美国Genentech公司于2000年推出了PLGA缓释微球产品(Nutropin)。该剂型是将rhGH与二价锌离子结合形成稳定的Zn-rhGH二聚体复合物，并采用喷雾冷冻干燥法制得，肌肉注射后可缓释2周至1个月。邱利炎等人在中国专利CN102302455B中，以聚乙二醇-聚乳酸嵌段共聚物，聚乙二醇-聚乳酸羟基乳酸嵌段共聚物为高分子材料，通过油包油包水复乳的方法制备了一种包载水溶性药物（包括重组人生长激素）的微囊。马光辉等人在中国专利CN102370630A中，同样以聚乙二醇-聚乳酸嵌段共聚物，聚乙二醇-聚乳酸羟基乳酸嵌段共聚物为高分子材料，通过水包油包水复乳的方法制备了重组人生长激素缓释药物微囊。以上所述的各种聚合物，均为单臂聚乙二醇-聚酯类线性聚合物为载体材料。聚合物微囊具有类似脂质体的双层结构，两个嵌段共聚物分子成对地规则排列，微囊壁由疏水链呈规则聚集，且由分别向壁内和壁外伸展的亲水链所稳定。聚合物微囊具有高度的稳定性，因而在药物传输方面有着广泛的应用。Drug sustained-release preparations are another effective strategy to effectively protect the long-term effect of protein drugs in vivo, mainly including liposomes (CN102188364A, CN101926770B), hydrogels (CN102989034A) and microcapsules (CN102302455B, CN102370630A). In 2000, Genentech Corporation of the United States launched the PLGA sustained-release microsphere product (Nutropin). The dosage form combines rhGH with divalent zinc ions to form a stable Zn-rhGH dimer complex, and is prepared by spray freeze-drying method, and can be sustained release for 2 weeks to 1 month after intramuscular injection. In the Chinese patent CN102302455B, Qiu Liyan et al. used polyethylene glycol-polylactic acid block copolymer and polyethylene glycol-polylactic acid hydroxylactic acid block copolymer as polymer materials, through the method of oil-in-oil-in-water re-emulsion A microcapsule loaded with water-soluble drugs (including recombinant human growth hormone) was prepared. In the Chinese patent CN102370630A, Ma Guanghui et al. also use polyethylene glycol-polylactic acid block copolymer and polyethylene glycol-polylactic acid hydroxylactic acid block copolymer as polymer materials, through water-in-oil-in-water reemulsion The recombinant human growth hormone sustained-release drug microcapsules were prepared by the method. The various polymers mentioned above are single-arm polyethylene glycol-polyester linear polymers as carrier materials. Polymer microcapsules have a bilayer structure similar to liposomes, two block copolymer molecules are regularly arranged in pairs, the microcapsule walls are regularly aggregated by hydrophobic chains, and hydrophilic The chain is stabilized. Polymer microcapsules have a high degree of stability, so they have a wide range of applications in drug delivery.
但是，以单臂聚乙二醇-聚酯类线性聚合物为载体材料制备缓释药物微囊，组成上含有聚乙烯醇等乳化剂为辅料，制备方法上需要首先形成复乳，使得制备工艺复杂，稳定性弱，重复性差，生物利用度低。重组人生长激素缓释药物微囊中重组人生长激素含量低。尤其是包封率不高，Guanghui Ma等人报道的以单臂聚乙二醇-聚乳酸包载重组人生长激素的缓释药物微囊，其包封率在65%以下（Journal of Materials Chemistry, 2011, 21, 12691）。However, the preparation of slow-release drug microcapsules with single-armed polyethylene glycol-polyester linear polymer as carrier material contains polyvinyl alcohol and other emulsifiers as auxiliary materials, and the preparation method needs to form double emulsion first, so that the preparation process Complex, weak stability, poor repeatability, low bioavailability. The content of recombinant human growth hormone in the recombinant human growth hormone slow-release medicine microcapsule is low. Especially the encapsulation efficiency is not high, Guanghui Ma et al. reported that the sustained-release drug microcapsules loaded with recombinant human growth hormone with single-arm polyethylene glycol-polylactic acid had an encapsulation efficiency below 65% (Journal of Materials Chemistry , 2011, 21, 12691).
发明内容Contents of the invention
为了解决已有技术存在的问题，本发明提供一种包载重组人生长激素缓释药物微囊。该包载重组人生长激素缓释药物微囊具有稳定性好，粒径均一，包裹效率高，生物活性高，制备工艺简单，重复性好的优点。In order to solve the problems in the prior art, the present invention provides a drug microcapsule loaded with recombinant human growth hormone for sustained release. The recombinant human growth hormone sustained-release medicine microcapsule has the advantages of good stability, uniform particle size, high encapsulation efficiency, high biological activity, simple preparation process and good repeatability.
本发明提供的一种包载重组人生长激素的缓释药物微囊，其组成如下：多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物载体与重组人生长激素的重量百分比为54-19%：46－81%；A slow-release drug microcapsule carrying recombinant human growth hormone provided by the present invention is composed as follows : a multi-armed polyethylene glycol-polyester block polymer is used as a carrier to carry recombinant human growth hormone; The weight percentages of the diol-polyester block polymer carrier and the recombinant human growth hormone are 54-19%: 46-81%;
所述的多臂聚乙二醇-聚酯嵌段聚合物为多臂聚乙二醇-聚乳酸嵌段聚合物，或多臂聚乙二醇-聚己内酯嵌段聚合物；优选四臂聚乙二醇2000-聚乳酸4000、四臂聚乙二醇2000-聚乳酸7000、四臂聚乙二醇2000-聚乳酸10000、三臂聚乙二醇2000-聚乳酸7000、六臂聚乙二醇2000-聚乳酸7000、八臂聚乙二醇2000-聚乳酸7000。The multi-arm polyethylene glycol-polyester block polymer is a multi-arm polyethylene glycol-polylactic acid block polymer, or a multi-arm polyethylene glycol-polycaprolactone block polymer; preferably four Arm polyethylene glycol2000 -polylactic acid4000 , four-arm polyethylene glycol2000 -polylactic acid7000 , four-arm polyethylene glycol2000 -polylactic acid10000 , three-arm polyethylene glycol2000 -polylactic acid7000 , six-arm poly Ethylene glycol2000 -polylactic acid7000 , eight-armed polyethylene glycol2000 -polylactic acid7000 .
本发明提供的包载重组人生长激素的缓释药物微囊的制备方法，包括以下步骤：The preparation method of the sustained-release drug microcapsules loaded with recombinant human growth hormone provided by the present invention comprises the following steps:
（1）所述的缓释药物微囊的制法如下：(1) The preparation method of the sustained-release drug microcapsules is as follows:
聚合物空心微囊的制备：将多臂聚乙二醇-聚酯嵌段聚合物置于反应器中，配置成浓度为0.5-2mg/mL的氯仿溶液，搅拌8-10小时，使此溶液中的氯仿蒸发形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；在持续搅拌条件下，向上述薄膜中加入去离子水，氯仿与去离子水体积比为1:1-4，多臂聚乙二醇-聚酯嵌段聚合物的质量mg与去离子水体积mL的比为1:2-2:1，继续搅拌24小时，冻干，得到聚合物空心微囊的冻干粉；Preparation of polymer hollow microcapsules: put the multi-arm polyethylene glycol-polyester block polymer in a reactor, configure a chloroform solution with a concentration of 0.5-2 mg/mL, stir for 8-10 hours, and make the solution The chloroform evaporated to form a polymer film; further, the reactor was placed on a rotary evaporator, and at room temperature, the chloroform was removed with a vacuum of 0.09 MPa; under continuous stirring conditions, deionized water and chloroform were added to the above film. The volume ratio with deionized water is 1:1-4, the ratio of the mass mg of multi-arm polyethylene glycol-polyester block polymer to the volume mL of deionized water is 1:2-2:1, and continue to stir for 24 hours , freeze-dried to obtain a freeze-dried powder of polymer hollow microcapsules;
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为1-5 mg/mL的重组人生长激素溶液；将聚合物空心微囊的冻干粉加入到反应器中，以0.5 mL/min的速率向反应器中加入上述配置的重组人生长激素溶液，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:１至1:５，持续搅拌24小时，然后，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。(2) Preparation of sustained-release drug microcapsules loaded with recombinant human growth hormone: prepare recombinant human growth hormone solution with a concentration of 1-5 mg/mL in phosphate buffer solution with pH=7.4; freeze polymer hollow microcapsules The dry powder is added into the reactor, and the above-mentioned recombinant human growth hormone solution is added to the reactor at a rate of 0.5 mL/min. The weight ratio of the lyophilized powder of the polymer hollow microcapsules to the recombinant human growth hormone is 1:1 to 1:5, continuously stirred for 24 hours, then centrifuged at 13,000 r/min at 4°C to remove recombinant human growth hormone not encapsulated in polymer microcapsules, and freeze-dried to obtain the target product.
重组人生长激素缓释药物微囊制备过程中，重组人生长激素的包封率采用紫外分光光度法测量，并按照以下公式进行计算：During the preparation of recombinant human growth hormone sustained-release drug microcapsules, the encapsulation efficiency of recombinant human growth hormone was measured by ultraviolet spectrophotometry and calculated according to the following formula:
包封率 = ×100％Encapsulation rate = ×100%
其中，rhGHtotal为步骤（2）中投入的重组人生长激素的总量，rhGHfree为步骤（2）中离心后上清液中的重组人生长激素的总量。本发明的包载重组人生长激素的缓释药物微囊的包封率达91%。Wherein, rhGHtotal is the total amount of recombinant human growth hormone input in step (2), and rhGHfree is the total amount of recombinant human growth hormone in the supernatant after centrifugation in step (2). The encapsulation efficiency of the sustained-release drug microcapsules carrying recombinant human growth hormone of the present invention reaches 91%.
重组人生长激素缓释药物微囊的粒径和形貌用透射电子显微镜表征。The particle size and morphology of recombinant human growth hormone sustained-release drug microcapsules were characterized by transmission electron microscopy.
本发明的包载重组人生长激素的缓释药物微囊的平均粒径范围在60-200 nm之间。The average particle size range of the slow-release drug microcapsules carrying recombinant human growth hormone of the present invention is between 60-200 nm.
本发明的包载重组人生长激素的缓释药物微囊的载药量：每mg缓释药物微囊中重组人生长激素的含量在0.46－0.81 mg。The drug loading capacity of the sustained-release drug microcapsules carrying recombinant human growth hormone of the present invention: the content of recombinant human growth hormone in each mg of sustained-release drug microcapsules is 0.46-0.81 mg.
有益效果：本发明的包载重组人生长激素的缓释药物微囊，以多臂聚乙二醇-聚酯嵌段聚合物为载体材料，相比于单臂聚乙二醇-聚酯嵌段聚合物，多臂聚合物具有较小的流体力学体积和较低的黏度，而其降解速率却比线性聚合物要大，更有利于药物的负载和输送。多臂聚乙二醇-聚酯嵌段共聚物因其具有良好的药物负载能力和独特的两亲性，可防止蛋白质的吸附并躲避网状内皮系统的识别与吞噬，以及易通过生理屏障等优点，赋予了本发明制备的重组人生长激素缓释药物微囊更好的生物利用度。另一方面，本发明通过大分子自组装形成缓释药物微囊，不需要复杂的复乳形成过程，制备工艺简单，微囊的稳定性好。本发明制备的产品的稳定性好，粒径均一。本发明的包载重组人生长激素的缓释药物微囊的包封率达到91%。重组人生长激素缓释药物微囊中重组人生长激素含量高达46－81%，生物活性高，重复性好，适用于大规模生产。Beneficial effects: the slow-release drug microcapsules loaded with recombinant human growth hormone of the present invention use multi-arm polyethylene glycol-polyester block polymer as carrier material, compared with single-arm polyethylene glycol-polyester block polymer Multi-arm polymers have smaller hydrodynamic volume and lower viscosity, but their degradation rate is higher than that of linear polymers, which is more conducive to drug loading and delivery. Because of its good drug loading capacity and unique amphiphilicity, the multi-arm polyethylene glycol-polyester block copolymer can prevent the adsorption of proteins, avoid the recognition and phagocytosis of the reticuloendothelial system, and easily pass through physiological barriers, etc. The advantage is that the recombinant human growth hormone sustained-release drug microcapsule prepared by the invention has better bioavailability. On the other hand, the present invention forms slow-release drug microcapsules through self-assembly of macromolecules, does not require complex double emulsion formation process, has simple preparation process, and has good stability of the microcapsules. The product prepared by the invention has good stability and uniform particle size. The encapsulation efficiency of the sustained-release drug microcapsules carrying recombinant human growth hormone of the present invention reaches 91%. The content of recombinant human growth hormone in microcapsules of recombinant human growth hormone sustained-release medicine is as high as 46-81%, with high biological activity and good repeatability, and is suitable for large-scale production.
附图说明Description of drawings
图1.实施例1的包载重组人生长激素缓释药物微囊的透射电镜照片；Fig. 1. the transmission electron micrograph of the microcapsules of slow-release drug loaded with recombinant human growth hormone of embodiment 1;
图2.实施例2的包载重组人生长激素缓释药物微囊的透射电镜照片；Fig. 2. the transmission electron micrograph of the slow-release drug microcapsules loaded with recombinant human growth hormone of embodiment 2;
图3.实施例3的包载重组人生长激素缓释药物微囊的透射电镜照片；Fig. 3. the transmission electron micrograph of the slow-release drug microcapsule loaded with recombinant human growth hormone of embodiment 3;
图4.实施例4的包载重组人生长激素缓释药物微囊的透射电镜照片；Fig. 4. the transmission electron micrograph of the slow-release drug microcapsule loaded with recombinant human growth hormone of embodiment 4;
图5.实施例5的包载重组人生长激素缓释药物微囊的透射电镜照片；Fig. 5. transmission electron micrograph of the slow-release drug microcapsules loaded with recombinant human growth hormone of embodiment 5;
图6.实施例6的包载重组人生长激素缓释药物微囊的透射电镜照片；Fig. 6. transmission electron micrograph of the slow-release drug microcapsules loaded with recombinant human growth hormone of embodiment 6;
图7.实施例7的包载重组人生长激素缓释药物微囊的透射电镜照片。Fig. 7. Transmission electron micrograph of microcapsules loaded with recombinant human growth hormone slow-release drug in Example 7.
具体实施方式Detailed ways
实施例1 一种包载重组人生长激素缓释药物微囊，多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物为四臂聚乙二醇2000-聚乳酸4000嵌段聚合物；四臂聚乙二醇2000-聚乳酸4000嵌段聚合物载体与重组人生长激素的重量百分比为５４％：４６％；Example 1 A slow-release drug microcapsule loaded with recombinant human growth hormone, the multi-armed polyethylene glycol-polyester block polymer is used as a carrier to carry recombinant human growth hormone; the multi-armed polyethylene glycol-polyester block polymer The block polymer is a four-arm polyethylene glycol2000 -polylactic acid4000 block polymer; the weight percentage of the four-arm polyethylene glycol2000 -polylactic acid4000 block polymer carrier and recombinant human growth hormone is 54%: 46% ;
所述的缓释药物微囊制法，步骤和条件如下：Described sustained-release drug microcapsule preparation method, steps and conditions are as follows:
（1）聚合物空心微囊的制备：取四臂聚乙二醇2000-聚乳酸4000嵌段聚合物10mg置于反应器中，加入20mL氯仿，多臂聚乙二醇-聚酯嵌段聚合物的质量mg与氯仿的体积mL的比为1：2，形成0.5mg/mL的氯仿溶液，搅拌8小时，使此溶液中的氯仿蒸发以形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；在持续搅拌条件下加入20 mL去离子水，氯仿与去离子水体积比为1:1，并继续搅拌24小时，形成聚合物空心微囊，冻干，得到聚合物空心微囊的冻干粉。(1) Preparation of polymer hollow microcapsules: Take 10 mg of the four-armed polyethylene glycol2000 -polylactic acid4000 block polymer in a reactor, add 20 mL of chloroform, and polymerize the multi-armed polyethylene glycol-polyester block The ratio of the mass mg of the substance to the volume mL of chloroform is 1:2 to form a 0.5 mg/mL chloroform solution and stir for 8 hours to evaporate the chloroform in the solution to form a polymer film; further, the reactor is placed in a rotating On the evaporator, at room temperature, remove chloroform with a vacuum of 0.09 MPa; add 20 mL of deionized water under continuous stirring, the volume ratio of chloroform to deionized water is 1:1, and continue stirring for 24 hours to form a polymer The hollow microcapsules are lyophilized to obtain the freeze-dried powder of polymer hollow microcapsules.
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为1 mg/mL的重组人生长激素溶液，将聚合物空心微囊的冻干粉20 mg加入到反应器中，然后以0.5 mL/min的速度加入上述重组人生长激素溶液20 mL，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:1,持续搅拌24小时，进一步，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。(2) Preparation of slow-release drug microcapsules loaded with recombinant human growth hormone: Prepare recombinant human growth hormone solution with a concentration of 1 mg/mL in phosphate buffer solution with pH = 7.4, and freeze-dried polymer hollow microcapsules 20 mg was added to the reactor, and then 20 mL of the above-mentioned recombinant human growth hormone solution was added at a rate of 0.5 mL/min, and the weight ratio of the lyophilized powder of the polymer hollow microcapsules to the recombinant human growth hormone was 1:1 , and continued to stir for 24 hours, and further, centrifuged at a speed of 13000 r/min at 4°C to remove recombinant human growth hormone not encapsulated in the polymer microcapsules, and freeze-dried to obtain the target product.
实施例1的理化性能如表1所示。The physical and chemical properties of Example 1 are shown in Table 1.
表1. 实施例1的理化性能Table 1. The physical and chemical properties of embodiment 1
实施例2：一种包载重组人生长激素缓释药物微囊，多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物为四臂聚乙二醇2000-聚乳酸7000嵌段聚合物；四臂聚乙二醇2000-聚乳酸7000嵌段聚合物载体与重组人生长激素的重量百分比为43%:57%。Example 2: A slow-release drug microcapsule loaded with recombinant human growth hormone, multi-armed polyethylene glycol-polyester block polymer as carrier, loaded with recombinant human growth hormone; multi-armed polyethylene glycol-polyester The block polymer is a four-arm polyethylene glycol2000 -polylactic acid7000 block polymer; the weight percentage of the four-arm polyethylene glycol2000 -polylactic acid7000 block polymer carrier and recombinant human growth hormone is 43%:57 %.
所述的缓释药物微囊制法，步骤和条件如下：Described sustained-release drug microcapsule preparation method, steps and conditions are as follows:
（1）聚合物空心微囊的制备：取四臂聚乙二醇2000-聚乳酸7000嵌段聚合物10mg置于反应器中，加入5mL氯仿，四臂聚乙二醇2000-聚乳酸7000嵌段聚合物的质量mg与氯仿的体积mL的比为2：1，形成2mg/mL的氯仿溶液，搅拌8小时，使此溶液中的氯仿蒸发以形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；在持续搅拌条件下加入20 mL去离子水，氯仿与去离子水体积比为1:4，并继续搅拌24小时，形成聚合物空心微囊，冻干，得到聚合物空心微囊的冻干粉。(1) Preparation of polymer hollow microcapsules: Take 10 mg of four-armed polyethylene glycol2000 -polylactic acid7000 block polymer and place it in a reactor, add 5 mL of chloroform, and four-armed polyethylene glycol2000 -polylactic acid7000 block polymer The ratio of the mass mg of polymer to the volume mL of chloroform is 2:1, forming a 2 mg/mL chloroform solution, stirred for 8 hours, and evaporated the chloroform in the solution to form a polymer film; further, the reactor was placed in On a rotary evaporator, remove chloroform with a vacuum of 0.09 MPa at room temperature; add 20 mL of deionized water under continuous stirring, the volume ratio of chloroform to deionized water is 1:4, and continue stirring for 24 hours to form The polymer hollow microcapsule is freeze-dried to obtain the freeze-dried powder of the polymer hollow microcapsule.
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为1 mg/mL的重组人生长激素溶液。将得到的聚合物空心微囊的冻干粉20 mg加入到反应器中，然后以0.5 mL/min的速度加入上述重组人生长激素溶液30 mL，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:1.5,持续搅拌24小时。进一步，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。(2) Preparation of slow-release drug microcapsules loaded with recombinant human growth hormone: Prepare recombinant human growth hormone solution with a concentration of 1 mg/mL in phosphate buffer solution with pH=7.4. Add 20 mg of the freeze-dried powder of the obtained polymer hollow microcapsules into the reactor, and then add 30 mL of the above-mentioned recombinant human growth hormone solution at a speed of 0.5 mL/min, the freeze-dried powder of the polymer hollow microcapsules The weight ratio to recombinant human growth hormone is 1:1.5, and the stirring is continued for 24 hours. Further, at a speed of 13000 r/min, at 4°C, centrifuge to remove the recombinant human growth hormone not encapsulated in the polymer microcapsules, and freeze-dry to obtain the target product.
实施例2的理化性能如表2所示。The physical and chemical properties of embodiment 2 are shown in table 2.
表2. 实施例2的理化性能Table 2. The physical and chemical properties of embodiment 2
实施例3：一种包载重组人生长激素缓释药物微囊，多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物为四臂聚乙二醇2000-聚乳酸10000嵌段聚合物；四臂聚乙二醇2000-聚乳酸10000嵌段聚合物载体与重组人生长激素的重量百分比为30%:70%。Example 3: A slow-release drug microcapsule loaded with recombinant human growth hormone, multi-armed polyethylene glycol-polyester block polymer as carrier, loaded with recombinant human growth hormone; multi-armed polyethylene glycol-polyester The block polymer is a four-arm polyethylene glycol2000 -polylactic acid10000 block polymer; the weight percentage of the four-arm polyethylene glycol2000 -polylactic acid10000 block polymer carrier and recombinant human growth hormone is 30%:70 %.
所述的缓释药物微囊制法，步骤和条件如下：Described sustained-release drug microcapsule preparation method, steps and conditions are as follows:
（1）聚合物空心微囊的制备：取四臂聚乙二醇2000-聚乳酸10000嵌段聚合物10mg置于反应器中，加入10mL氯仿，四臂聚乙二醇2000-聚乳酸10000嵌段聚合物的质量mg与氯仿的体积mL的比为1：1，形成1mg/mL的氯仿溶液，搅拌10小时，使此溶液中的氯仿蒸发以形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；在持续搅拌条件下加入20 mL去离子水，氯仿与去离子水体积比为1:2，并继续搅拌24小时，形成聚合物空心微囊，冻干，得到聚合物空心微囊的冻干粉。(1) Preparation of polymer hollow microcapsules: Take 10 mg of the four-arm polyethylene glycol2000 -polylactic acid10000 block polymer and place it in a reactor, add 10 mL of chloroform, and four-arm polyethylene glycol2000 -polylactic acid10000 block polymer The ratio of the mass mg of the segment polymer to the volume mL of chloroform is 1:1 to form a 1 mg/mL chloroform solution and stir for 10 hours to evaporate the chloroform in the solution to form a polymer film; further, place the reactor in On a rotary evaporator, remove chloroform with a vacuum of 0.09 MPa at room temperature; add 20 mL of deionized water under continuous stirring, the volume ratio of chloroform to deionized water is 1:2, and continue stirring for 24 hours to form The polymer hollow microcapsule is freeze-dried to obtain the freeze-dried powder of the polymer hollow microcapsule.
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为3 mg/mL的重组人生长激素溶液，将得到的聚合物空心微囊的冻干粉20 mg加入到反应器中，然后以0.5 mL/min的速度加入上述重组人生长激素溶液20 mL，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:3,持续搅拌24小时。进一步，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。(2) Preparation of slow-release drug microcapsules loaded with recombinant human growth hormone: Prepare recombinant human growth hormone solution with a concentration of 3 mg/mL in phosphate buffer solution with pH = 7.4, and freeze the obtained polymer hollow microcapsules. 20 mg of dry powder was added to the reactor, and then 20 mL of the above-mentioned recombinant human growth hormone solution was added at a rate of 0.5 mL/min. The weight ratio of the freeze-dried powder of the polymer hollow microcapsules to recombinant human growth hormone was 1 :3, keep stirring for 24 hours. Further, at a speed of 13000 r/min, at 4°C, centrifuge to remove the recombinant human growth hormone not encapsulated in the polymer microcapsules, and freeze-dry to obtain the target product.
实施例3的理化性能如表3所示。The physical and chemical properties of embodiment 3 are shown in table 3.
表3. 实施例3的理化性能Table 3. The physical and chemical properties of embodiment 3
实施例4：一种包载重组人生长激素缓释药物微囊，多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物为三臂聚乙二醇2000-聚乳酸7000；三臂聚乙二醇2000-聚乳酸7000载体与重组人生长激素的重量百分比为19%:81%。Example 4: A slow-release drug microcapsule loaded with recombinant human growth hormone, multi-armed polyethylene glycol-polyester block polymer as carrier, loaded with recombinant human growth hormone; multi-armed polyethylene glycol-polyester The block polymer is three-arm polyethylene glycol2000 -polylactic acid7000 ; the weight percentage of three-arm polyethylene glycol2000 -polylactic acid7000 carrier and recombinant human growth hormone is 19%:81%.
所述的缓释药物微囊制法，步骤和条件如下：Described sustained-release drug microcapsule preparation method, steps and conditions are as follows:
（1）聚合物空心微囊的制备：取三臂聚乙二醇2000-聚乳酸7000嵌段聚合物10mg置于反应器中，加入10mL氯仿，多臂聚乙二醇-聚酯嵌段聚合物的质量mg与氯仿的体积mL的比为1：1，形成1mg/mL的氯仿溶液，搅拌10小时，使此溶液中的氯仿蒸发以形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；在持续搅拌条件下加入20mL去离子水，氯仿与去离子水体积比为1:2，并继续搅拌24小时，形成聚合物空心微囊，冻干，得到聚合物空心微囊的冻干粉。(1) Preparation of polymer hollow microcapsules: Take 10 mg of the three-arm polyethylene glycol2000 -polylactic acid7000 block polymer in a reactor, add 10 mL of chloroform, and polymerize the multi-arm polyethylene glycol-polyester block The ratio of the mass mg of the substance to the volume mL of chloroform is 1:1, forming a 1 mg/mL chloroform solution, stirring for 10 hours, and evaporating the chloroform in the solution to form a polymer film; further, placing the reactor in a rotary evaporator At room temperature, remove chloroform with a vacuum of 0.09 MPa; add 20 mL of deionized water under continuous stirring, the volume ratio of chloroform to deionized water is 1:2, and continue stirring for 24 hours to form a polymer hollow The microcapsules are freeze-dried to obtain the freeze-dried powder of polymer hollow microcapsules.
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为5 mg/mL的重组人生长激素溶液，将得到的聚合物空心微囊的冻干粉20 mg加入到反应器中，然后以0.5 mL/min的速度加入上述重组人生长激素溶液20 mL，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:5,持续搅拌24小时，进一步，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。(2) Preparation of slow-release drug microcapsules loaded with recombinant human growth hormone: Prepare recombinant human growth hormone solution with a concentration of 5 mg/mL in phosphate buffer solution with pH = 7.4, and freeze the obtained polymer hollow microcapsules. 20 mg of dry powder was added to the reactor, and then 20 mL of the above-mentioned recombinant human growth hormone solution was added at a rate of 0.5 mL/min. The weight ratio of the freeze-dried powder of the polymer hollow microcapsules to recombinant human growth hormone was 1 :5, continuously stirred for 24 hours, further, with the speed of 13000 r/min, under the condition of 4 ℃, centrifuged to remove the recombinant human growth hormone not contained in the polymer microcapsules, freeze-dried to obtain the target product.
实施例4的理化性能如表4所示。The physical and chemical properties of embodiment 4 are shown in table 4.
表4. 实施例4的理化性能Table 4. The physical and chemical properties of embodiment 4
实施例5：一种包载重组人生长激素缓释药物微囊，多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物为六臂聚乙二醇2000-聚乳酸7000嵌段聚合物；六臂聚乙二醇2000-聚乳酸7000嵌段聚合物载体与重组人生长激素的重量百分比为28%:72%。Example 5: A slow-release drug microcapsule loaded with recombinant human growth hormone, multi-armed polyethylene glycol-polyester block polymer as carrier, loaded with recombinant human growth hormone; multi-armed polyethylene glycol-polyester The block polymer is a six-arm polyethylene glycol2000 -polylactic acid7000 block polymer; the weight percentage of the six-arm polyethylene glycol2000 -polylactic acid7000 block polymer carrier and recombinant human growth hormone is 28%:72 %.
所述的缓释药物微囊制法，步骤和条件如下：Described sustained-release drug microcapsule preparation method, steps and conditions are as follows:
（1）聚合物空心微囊的制备：取六臂聚乙二醇2000-聚乳酸7000嵌段聚合物10mg置于反应器中，加入10mL氯仿，多臂聚乙二醇-聚酯嵌段聚合物的质量mg与氯仿的体积mL的比为1：1，形成1mg/mL的氯仿溶液，搅拌9小时，使此溶液中的氯仿蒸发以形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；在持续搅拌条件下加入20 mL去离子水，氯仿与去离子水体积比为1:2，并继续搅拌24小时，形成聚合物空心微囊，冻干，得到聚合物空心微囊的冻干粉。(1) Preparation of polymer hollow microcapsules: Take 10 mg of six-arm polyethylene glycol2000 -polylactic acid7000 block polymer in a reactor, add 10 mL of chloroform, and polymerize the multi-arm polyethylene glycol-polyester block The ratio of the mass mg of the substance to the volume mL of chloroform is 1:1 to form a 1 mg/mL chloroform solution and stir for 9 hours to evaporate the chloroform in the solution to form a polymer film; further, the reactor is placed in a rotary evaporator At room temperature, remove chloroform with a vacuum of 0.09 MPa; add 20 mL of deionized water under continuous stirring, the volume ratio of chloroform to deionized water is 1:2, and continue stirring for 24 hours to form a polymer The hollow microcapsules are freeze-dried to obtain the freeze-dried powder of the polymer hollow microcapsules.
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为3 mg/mL的重组人生长激素溶液。将得到的聚合物空心微囊的冻干粉20 mg加入到反应器中，然后以0.5 mL/min的速度加入上述重组人生长激素溶液20 mL，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:3,持续搅拌24小时。进一步，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。实施例5的理化性能如表5所示。(2) Preparation of slow-release drug microcapsules loaded with recombinant human growth hormone: Prepare recombinant human growth hormone solution with a concentration of 3 mg/mL in phosphate buffer solution with pH=7.4. Add 20 mg of the freeze-dried powder of the obtained polymer hollow microcapsules into the reactor, and then add 20 mL of the above-mentioned recombinant human growth hormone solution at a speed of 0.5 mL/min, the freeze-dried powder of the polymer hollow microcapsules The weight ratio to recombinant human growth hormone is 1:3, and the stirring is continued for 24 hours. Further, at a speed of 13000 r/min, under the condition of 4°C, the recombinant human growth hormone not encapsulated in the polymer microcapsules was removed by centrifugation, and then freeze-dried to obtain the target product. The physical and chemical properties of Example 5 are shown in Table 5.
表5. 实施例5的理化性能Table 5. The physical and chemical properties of embodiment 5
实施例6：一种包载重组人生长激素缓释药物微囊，多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物为八臂聚乙二醇2000-聚乳酸7000嵌段聚合物；八臂聚乙二醇2000-聚乳酸7000嵌段聚合物载体与重组人生长激素的重量百分比为27%:73%。Example 6: A slow-release drug microcapsule loaded with recombinant human growth hormone, multi-armed polyethylene glycol-polyester block polymer as carrier, loaded with recombinant human growth hormone; multi-armed polyethylene glycol-polyester The block polymer is an eight-arm polyethylene glycol2000 -polylactic acid7000 block polymer; the weight percentage of the eight-arm polyethylene glycol2000 -polylactic acid7000 block polymer carrier and recombinant human growth hormone is 27%: 73 %.
所述的缓释药物微囊制法，步骤和条件如下：Described sustained-release drug microcapsule preparation method, steps and conditions are as follows:
（1）聚合物空心微囊的制备：取八臂聚乙二醇2000-聚乳酸7000嵌段聚合物10mg置于反应器中，加入10mL氯仿，多臂聚乙二醇-聚酯嵌段聚合物的质量mg与氯仿的体积mL的比为1：1，形成1mg/mL的氯仿溶液，搅拌10小时，使此溶液中的氯仿蒸发以形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；最后，在持续搅拌条件下加入20 mL去离子水，氯仿与去离子水体积比为1:2，并继续搅拌24小时，形成聚合物空心微囊，冻干，得到聚合物空心微囊的冻干粉。(1) Preparation of polymer hollow microcapsules: Take 10 mg of eight-armed polyethylene glycol2000 -polylactic acid7000 block polymer in a reactor, add 10 mL of chloroform, and polymerize the multi-armed polyethylene glycol-polyester block The ratio of the mass mg of the substance to the volume mL of chloroform is 1:1, forming a 1 mg/mL chloroform solution, stirring for 10 hours, and evaporating the chloroform in the solution to form a polymer film; further, placing the reactor in a rotary evaporator At room temperature, remove chloroform with a vacuum of 0.09 MPa; finally, add 20 mL of deionized water under constant stirring, the volume ratio of chloroform to deionized water is 1:2, and continue stirring for 24 hours to form The polymer hollow microcapsule is freeze-dried to obtain the freeze-dried powder of the polymer hollow microcapsule.
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为3 mg/mL的重组人生长激素溶液。将得到的聚合物空心微囊的冻干粉20 mg加入到反应器中，然后以0.5 mL/min的速度加入上述重组人生长激素溶液20 mL，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:3,持续搅拌24小时。进一步，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。(2) Preparation of slow-release drug microcapsules loaded with recombinant human growth hormone: Prepare recombinant human growth hormone solution with a concentration of 3 mg/mL in phosphate buffer solution with pH=7.4. 20 mg of the freeze-dried powder of the obtained polymer hollow microcapsules was added to the reactor, and then 20 mL of the above-mentioned recombinant human growth hormone solution was added at a speed of 0.5 mL/min, and the freeze-dried powder of the polymer hollow microcapsules The weight ratio to recombinant human growth hormone is 1:3, and the stirring is continued for 24 hours. Further, at a speed of 13000 r/min, under the condition of 4°C, the recombinant human growth hormone not encapsulated in the polymer microcapsules was removed by centrifugation, and then freeze-dried to obtain the target product.
实施例6的理化性能如表6所示。The physical and chemical properties of Example 6 are shown in Table 6.
表6. 实施例6的理化性能Table 6. The physical and chemical properties of embodiment 6
实施例7：一种包载重组人生长激素缓释药物微囊，多臂聚乙二醇-聚酯嵌段聚合物为载体，包载重组人生长激素；多臂聚乙二醇-聚酯嵌段聚合物为四臂聚乙二醇2000-聚己内酯7000嵌段聚合物；四臂聚乙二醇2000-聚己内酯7000嵌段聚合物载体与重组人生长激素的重量百分比为45%:55%。Example 7: A slow-release drug microcapsule loaded with recombinant human growth hormone, multi-armed polyethylene glycol-polyester block polymer as carrier, loaded with recombinant human growth hormone; multi-armed polyethylene glycol-polyester The block polymer is a four-arm polyethylene glycol2000 -polycaprolactone7000 block polymer; the weight percentage of the four-arm polyethylene glycol2000 -polycaprolactone7000 block polymer carrier and recombinant human growth hormone is 45%:55%.
所述的缓释药物微囊制法，步骤和条件如下：Described sustained-release drug microcapsule preparation method, steps and conditions are as follows:
（1）聚合物空心微囊的制备：取八臂聚乙二醇2000-聚乳酸7000嵌段聚合物10mg置于反应器中，加入10mL氯仿，多臂聚乙二醇-聚酯嵌段聚合物的质量mg与氯仿的体积mL的比为1:1，形成1mg/mL的氯仿溶液，搅拌10小时，使此溶液中的氯仿蒸发以形成聚合物薄膜；进一步，将反应器置于旋转蒸发器上，在室温下，以0.09 MPa的真空度除净氯仿；在持续搅拌条件下加入20 mL去离子水，氯仿与去离子水体积比为1:2，并继续搅拌24小时，形成聚合物空心微囊，冻干，得到聚合物空心微囊的冻干粉。(1) Preparation of polymer hollow microcapsules: Take 10 mg of eight-armed polyethylene glycol2000 -polylactic acid7000 block polymer in a reactor, add 10 mL of chloroform, and polymerize the multi-armed polyethylene glycol-polyester block The ratio of the mass mg of the substance to the volume mL of chloroform is 1:1 to form a 1 mg/mL chloroform solution, stir for 10 hours, and evaporate the chloroform in the solution to form a polymer film; further, place the reactor in a rotary evaporation At room temperature, remove chloroform with a vacuum of 0.09 MPa; add 20 mL of deionized water under continuous stirring, the volume ratio of chloroform to deionized water is 1:2, and continue stirring for 24 hours to form a polymer The hollow microcapsules are freeze-dried to obtain the freeze-dried powder of the polymer hollow microcapsules.
（2）包载重组人生长激素的缓释药物微囊的制备：以pH=7.4的磷酸缓冲溶液配置浓度为1 mg/mL的重组人生长激素溶液。将得到的聚合物空心微囊的冻干粉20 mg加入到反应器中，然后以0.5 mL/min的速度加入上述重组人生长激素溶液30 mL，所述的聚合物空心微囊的冻干粉与重组人生长激素的重量比为1:1.5,持续搅拌24小时。进一步，以13000 r/min的速度，4℃条件下，离心除去没有包载在聚合物微囊内的重组人生长激素，冻干，得到目标产物。(2) Preparation of slow-release drug microcapsules loaded with recombinant human growth hormone: Prepare recombinant human growth hormone solution with a concentration of 1 mg/mL in phosphate buffer solution with pH=7.4. Add 20 mg of the freeze-dried powder of the obtained polymer hollow microcapsules into the reactor, and then add 30 mL of the above-mentioned recombinant human growth hormone solution at a speed of 0.5 mL/min, the freeze-dried powder of the polymer hollow microcapsules The weight ratio to recombinant human growth hormone is 1:1.5, and the stirring is continued for 24 hours. Further, at a speed of 13000 r/min, under the condition of 4°C, the recombinant human growth hormone not encapsulated in the polymer microcapsules was removed by centrifugation, and then freeze-dried to obtain the target product.
实施例7的理化性能如表7所示。The physical and chemical properties of Example 7 are shown in Table 7.
表7. 实施例7的理化性能Table 7. The physical and chemical properties of embodiment 7
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| CN201410238630.2ACN103976982A (en) | 2014-06-03 | 2014-06-03 | Recombinant human growth hormone entrapped sustained-release drug microcapsules |
| Application Number | Priority Date | Filing Date | Title |
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| CN201410238630.2ACN103976982A (en) | 2014-06-03 | 2014-06-03 | Recombinant human growth hormone entrapped sustained-release drug microcapsules |
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| CN103976982Atrue CN103976982A (en) | 2014-08-13 |
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