CN1287798C - Method for preparing long acting progestational hormone injection embedded agent and use - Google Patents

Abstract

The present invention adopts biodegradable polymers, such as PLGA or PLA and the like as a framework material, and adopts gestagen-trenbolone as a model medicine to prepare a microsphere for injecting by a solvent evaporation method. The microsphere can be subcutaneously injected or injected in muscles, and medicine can slow release for more than 15 days. The framework material of the microsphere can be gradually degradated in human bodies to form intrinsic small molecule substances of organisms to be exhausted out of the human bodies. Besides the trenbolone microsphere injection is convenient to use, the trenbolone microsphere injection can obviously reduce adverse reactions, such as liver and kidney harm, etc., caused by the frequent oral administration of the existing conventional trenbolone preparation, namely tablets. Rat in vivo tests indicate that the novel trenbolone microsphere injection can effectively treat endometriosis, etc.

Description

Preparation method and application of long-acting progesterone injection implant

Technical field:

The invention relates to a preparation method and application of a long-acting progesterone injection implant.

Background technique:

Progestogens, such as gestrinone (Gestrinone, R2323, norethindrone, 18-methyltrienyl norethindrone) were originally moderate-strength progestogens with strong antiprogestogen and antiestrogenic activities. Experiments have proved that this product has anti-implantation and anti-early pregnancy effects, and it can also inhibit ovulation when taken orally in the early menstrual cycle. Its anti-implantation and anti-early pregnancy effects are related to changing the thickness of cervical mucus, interfering with the development of endometrium, affecting the running speed of eggs and antagonizing endometrial progesterone receptors. Clinically, it was originally used as a family-visiting contraceptive or an after-event contraceptive.

In recent years, clinical studies at home and abroad have found that oral administration of certain progestogens, such as gestrinone, still has the effect of treating endometriosis and uterine fibroids, and the curative effect is reliable ( ^① Clin Exp Obstel Gynecol1996, 23(4): 198 -204), ( ^② Chinese Medical Journal (Taipei) 1996, 58(2): 89-96), ( ^③ Gynecol Obstel Invest 1997, 43(3): 192-194), ( ^④ J Med Assoc Thai 1999, 82 (1):9-14).

At present, the dosage form of clinical progesterone, such as gestrinone, is only one kind of tablet, the dosage is 1.5mg, 2.5mg. For the treatment of endometriosis, orally twice a week, 2.5 mg each time, a course of 6 months. Frequent dosing brings inconvenience to patients. Forgetting to take medication will have adverse effects on treatment. It should be noted that progesterone, such as gestrinone, is taken orally, and is metabolized and excreted by the liver and kidneys. Long-term use will cause certain damage to the human liver and kidneys (Edited by Chen Xinqian, Newly edited Pharmacology, 15th edition, People's Health Publishing Society, P597).

Invention content:

The technical problem to be solved in the present invention is to disclose the preparation method and application of a long-acting progesterone injection implant, so as to overcome the inconvenience caused by the frequent administration of existing dosage forms to patients and the certain damage to human liver and kidney. defect.

Technical scheme of the present invention:

The long-acting progesterone injection implant of the present invention is one of microparticles, microspheres, nanoparticles or nanospheres, including an effective amount of active ingredient progesterone and a biodegradable polymer as a skeleton material, and the microspheres have a diameter of For a round entity of 10-120 microns, the preferred weight percentage content of the progesterone is 1-70%.

Said progestin includes gestrinone, gestodene (Gestrodene), nomegestrol (Nomegestrol), pulmegestone (Promegestone), norethindrone (Norgestrol), megestrol (Megestrol), norgestrel (Norgestrel), levonorgestrel (Levonorgestrel), desogestrol (Desogestrol), norgestimate (Norgestimate), ketone oxime (NorehisteroneOxime), quingestrel (Quingestanod), or anordrin One of esters (Anordrin), etc., preferably gestrinone.

Said biodegradable polymers include polylactide-glycolide, polylactic acid, polyglycolic acid, polycaprolactone, polyanhydride, polyhydroxybutyrate-hydroxyvalerate copolymer, polypropylene dextran, One or more of polylactic acid-polyethylene glycol, preferably a polymer with a molecular weight ranging from 5,000 to 1,000,000, wherein the polylactide-glycolide monomer polymerization ratio is between 95:5 and 5:95 , the molecular weight is between 10,000-500,000, and the molecular weight of polylactic acid is between 5,000-300,000.

The present invention also relates to a pharmaceutical composition composed of a therapeutically effective amount of the above-mentioned preparation and a biocompatible and degradable carrier. The above-mentioned preparation can be conveniently prepared into subcutaneous or intramuscular injection, subcutaneous Implants such as tablets, films, sticks, pills.

The preparation of the present invention can be effectively used for treating diseases such as endometriosis and hysteromyoma. The preparation method of the long-acting progesterone microsphere injection of the present invention comprises the following steps:

Dissolve the progestin and biodegradable polymer in the organic solvent, after high-speed stirring, pour into the stirring aqueous solution containing the suspending agent, make an emulsion and place it at a temperature of 5-40°C, and evaporate the organic solvent , and then adopt conventional methods to collect microspheres from the mixture, which is the long-acting progesterone injection preparation of the present invention.

The content of the degradable polymer in the organic solvent is 1-500mg/ml;

Said organic solvent includes dichloromethane, chloroform, ethyl acetate, ether or one or more;

Said suspending agent includes polyvinyl alcohol, CMC, PVP, natural colloid, such as a kind of gelatin, gum arabic, etc., and the content of suspending agent in aqueous solution is 1～50mg/ml;

The volume ratio of the organic solvent containing the progesterone and the biodegradable polymer to the aqueous solution containing the suspending agent is 1:1-500 (v/v).

The method of using the preparation of the present invention is as follows: inject the sterile aqueous solution containing suspending agent into the prepared microspheres, mix and shake, then inject subcutaneously or intramuscularly, and the dosage is handled according to the indicated amount in the instructions of different pharmaceutical preparations.

The treatment of endometriosis, uterine fibroids and other symptoms can be given to patients requiring the above treatment at intervals of at least 15 days.

The drug contained within can be controlled and released for a long period of time, and the skeleton material can be automatically degraded into inherent non-toxic small molecular substances in the body, which are finally absorbed by the body.

After the injection is administered, the drug can be released slowly in the body for more than half a month. It can reduce the frequency of medication for patients, which can not only facilitate the treatment of patients, but also reduce the adverse reactions such as liver and kidney damage that may be caused by oral administration of tablets.

Detailed ways:

Example 1

Take 20mg gestrinone, 80mg polylactide-glycolide (lactide:glycolide=75:25), dissolve in 3ml dichloromethane, and drop it into 100ml under vigorous stirring (1000rpm) In 0.2% sodium carboxymethylcellulose aqueous solution, continue to stir for 5-10 minutes after dripping, then reduce the stirring speed to 300rpm, evaporate at 25°C for about 12h, centrifuge, pour off the supernatant, filter the precipitate under reduced pressure, and a small amount of distilled water Wash the precipitate, dry under reduced pressure at room temperature, and the diameter of the microspheres is 40-120 μm.

Example 2

Take 20mg gestrinone and 80mg polylactic acid, dissolve them in 1ml dichloromethane, add 5ml 0.5% PVA aqueous solution in a high-speed homogenizer, stir vigorously at 6000rpm for 2min, pour into 100ml 0.5% PVA aqueous solution, and Evaporate at 300 rpm at 25°C for about 12 hours, centrifuge, pour off the supernatant, filter the precipitate under reduced pressure, rinse the precipitate with a small amount of distilled water, and dry under reduced pressure at room temperature. The diameter of the microspheres is 30-80 μm.

Example 3

Inhibition test of rat estrous cycle:

Method: The rats with regular estrous cycle were divided into two groups, and the following corresponding injections were given on the day when the pictures of the vaginal secretions of the rats showed anaphase (white blood cells):

Negative control group: subcutaneous (s.c.) injection of vehicle (n=5).

The microsphere group of Example 1: single dose s.c. injection of microspheres with a release rate of 0.5 mg/week/kg, a particle size of 40-120 μm, and a gestrinone content of 25% in the microspheres (n=5).

After injection, regularly observe and record vaginal cell smears every day. Rat vaginal smears continue to show white blood cells, indicating that the estrous cycle is inhibited. If keratinocytes appear, it indicates that the estrous cycle begins to recover. The vaginal smear method can be used to judge the drug effect of the preparation. time.

Results: After s.c. injection of a single dose of microspheres 0.5mg/week/kg in rats, white blood cells appeared in the vaginal smears of five rats at 15 days, 16 days, 16 days, 15 days, and 15 days, followed by round cells and Keratinocytes. Explain that its estrous cycle has been continuously suppressed for 15 days. The negative control group was not suppressed.

Example 4

Efficacy of endometriosis in rats:

Method: refer to Jones method (Acta Endocrinol 1984, 106: 282-88). A rat model of endometriosis was established by surgical autotransplantation. A corner of the uterus of a female rat was cut off and transplanted to the abdominal wall. Four weeks later, the abdominal cavity was re-opened to check for ectopic intimal growth. Those with good growth of ectopic endometrium and yellow liquid contents at the inhibition site were successful in establishing the model. After the volume was measured, the animals were randomly divided into groups for the following treatments:

Negative control group: the back of the rats was injected s.c. with normal saline every day (divided into 10 groups, n=3 in each group). The microsphere group of embodiment 2: disposable s.c. injection of microspheres on the back of rats, dosage is 0.5 mg/week/kg, particle diameter 30～80 μ m, microsphere gestrinone content 25% (divided into 10 groups, each group n =3).

At the end of the second week, the rats were sacrificed under anesthesia, the abdominal cavity was opened, the volume of ectopic intima was measured, and the inhibition rate was calculated.

Inhibition rate%=(1-V ₂ /V ₁ )×100%

V ₁ : graft volume before treatment (mm ² )

V ₂ : graft volume after treatment (mm ² )

The results are shown in Table 1.

It can be seen from the experimental results that the volume of the negative control group increased from 61.52 to 63.15 before and after injection, and the inhibition rate was -2.65%, and the volume of the microsphere group decreased from 49.80 to 20.17 after injection, and the inhibition rate was 59.50%. (p<0.05), and there was no increase in body weight after administration.

Table 1. Gestrinone Microsphere Injection Inhibitory Test Results on Endometriosis in Rats group 1 2 3 4 5 6 7 8 9 10 11 12 empty pair 5.45 4.45 5.22 4.95 6.29 7.00 2.82 3.02 7.69 3.88 average value SD P value 5.01 4.42 5.14 5.04 4.72 5.27 2.83 3.02 6.89 2.88 1.69 2.36 2.21 2.56 2.07 2.90 2.16 1.64 3.32 2.05 predrug volume 46.14 46.42 59.30 63.87 61.46 106.98 17.24 14.96 175.91 22.91 61.52 48.59 2.82 3.78 6.21 3.72 5.29 7.19 5.94 5.22 4.87 5.99 2.89 4.36 6.21 4.99 4.99 6.78 5.14 5.09 5.76 5.37 1.09 4.36 1.62 1.00 3.36 2.89 2.94 2.05 1.19 1.94 Post-drug volume 8.88 71.86 62.47 18.56 88.69 140.88 89.94 54.47 33.38 62.40 63.15 38.56 volume factor 0.19 1.55 1.05 0.29 1.44 1.32 5.22 3.64 0.19 2.72 1.76 1.64 weight 246.60 249.30 251.10 266.60 215.00 276.70 267.60 240.40 274.40 258.50 254.62 18.51 R2323 group 5.71 5.99 5.08 5.69 6.40 5.44 6.10 3.90 4.60 4.79 average value SD P value 5.38 5.99 5.09 5.49 5.69 3.53 6.18 3.90 6.34 5.12 0.97 2.60 1.50 0.80 1.73 2.64 2.30 1.68 2.44 0.57 predrug volume 29.80 93.29 38.79 24.99 63.00 50.70 86.71 25.55 71.16 13.98 49.80 27.67 4.15 4.45 5.78 4.58 3.83 3.48 4.59 3.63 5.92 3.13 4.15 3.35 5.12 4.88 4.18 3.45 4.58 3.60 5.32 4.20 0.23 0.85 1.27 0.80 0.76 0.73 0.50 0.63 2.63 0.54 Post-drug volume 3.96 12.67 37.58 17.88 12.17 8.76 10.51 8.23 82.83 7.10 20.17 23.93 volume factor 0.13 0.14 0.97 0.72 0.19 0.17 0.12 0.32 1.16 0.51 0.44 0.38 0.0238029 weight 202.30 226.40 228.20 233.20 250.40 256.50 238.20 223.60 258.80 226.70 234.43 17.21

Claims (6)

1. A progesterone injection implant, characterized in that it includes the active ingredient progesterone and a biodegradable polymer as a skeleton material; The weight percentage of progesterone is 1-70%; said progesterone is gestrinone; The biodegradable polymer is polylactide-glycolide, the polylactide-glycolide monomer polymerization ratio is 75:25, and the molecular weight is 10,000-500,000. 2. The progesterone injection implant according to claim 1, characterized in that the preparation is one of microparticles, microspheres, nanoparticles or nanospheres. 3. The progesterone injection implant according to claim 2, characterized in that the microspheres are round entities with a diameter of 10-120 microns. 4. The method for preparing the progesterone injection implant according to claim 3, characterized in that it comprises the following steps: dissolving the progesterone and the biodegradable polymer in an organic solvent, and pouring it into the stirred aqueous solution containing a suspending agent , make an oil-in-water emulsion, evaporate the organic solvent, and then collect microspheres from the mixture by conventional methods, which is the preparation of the present invention. 5. The preparation method according to claim 4, characterized in that the content of the degradable polymer in the organic solvent is 1-500 mg/ml; Said organic solvent is selected from one or more of dichloromethane, chloroform, ethyl acetate or ether; Said suspending agent is selected from polyvinyl alcohol, carboxymethylcellulose, polyvinylpyrrolidone or natural colloid, and the content of suspending agent in the aqueous solution is 1～50mg/ml; The volume ratio of the organic solvent containing the progesterone and the biodegradable polymer to the aqueous solution containing the suspending agent is 1:1-500. 6. Use of the progesterone injection implant according to claim 1 in the preparation of medicines for treating endometriosis or uterine fibroids.