CN1562042A - Method for preparing long acting progestogen injection implant agent and use - Google Patents

Abstract

The invention discloses that the biodegradable polymer such as PLGA or PLA is used as a frame material, and the progesterone-gestrinone (R2323) is used as a model drug to prepare injectable microspheres through a solvent evaporation method. The microspheres can be injected subcutaneously or intramuscularly, and the drug can be released slowly for more than 15 days. The microsphere skeleton material can be gradually degraded into small molecular substances inherent in the body and excreted from the body. Gestrinone Microsphere Injection is not only convenient for medication, but also can significantly reduce adverse reactions such as liver and kidney damage caused by frequent oral administration of tablets, the current conventional dosage form of gestrinone. Tests in vivo on rats show that the new dosage form can effectively treat diseases such as endometriosis.

Description

Preparation method and application of long-acting progesterone injection implant

technical field

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

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 gestrinone still has the effect of treating endometriosis, uterine fibroids and other diseases, and the curative effect is reliable, such as literature ^① Clin Exp Obstel Gynecol 1996, 23(4): 198-204, ^② Chinese Medical Journal (Taipei) 1996, 58(2): 89-96, ^③ Gynecol Obstel Invest 1997, 43(3): 192-194) and ^④ J Med Assoc Thai 1999, 82(1): 9-14 reports.

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 administration of medicines brings a lot of inconvenience to patients, and forgetting medicines brings 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).

Contents of the invention

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

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 a therapeutically effective amount of the active ingredient progesterone and a biodegradable polymer as a skeleton material, and the microspheres have a diameter of It is a round entity of 10-120 microns, and 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 polymer includes polylactide-glycolide, polylactic acid, polyglycolic acid, polycaprolactone, polyanhydride, polyhydroxybutyrate-hydroxyvalerate copolymer, polypropylene dextran or 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 weight ratio of polylactide-glycolide monomer polymerization is 95:5 to 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 therapeutically effective dose of long-acting progesterone and a pharmaceutical composition composed of a biocompatible and degradable carrier. The above-mentioned preparation can also be prepared by methods known in the art such as molding, casting, extrusion, Film-spreading and other methods are made into tablets, films, rods, pills, etc. for subcutaneous implantation.

Said degradable carrier comprises one or its mixture in polylactide-glycolide, polyDL-lactic acid, or polyanhydride etc., the content of preferred long-acting progesterone in injection implant (w/w ) is: 1-70%;

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:

Pour the organic solvent containing progesterone and biodegradable polymer into the aqueous solution containing suspending agent to make an emulsion, evaporate the organic solvent at a temperature of 5-40°C, and then remove the organic solvent from the suspension by a conventional method. The microspheres are collected, sterilized by gamma-rays or packed in aseptically, and then the long-acting progesterone injection preparation of the present invention is obtained.

The content of the degradable polymer in the organic solvent is 1-1000mg/ml;

The content of progesterone in the organic solvent is 1-800mg/ml;

The content of the suspending agent in the aqueous solution is 1-100mg/ml;

The volume ratio of the organic solvent containing the progesterone and the biodegradable polymer to the aqueous solution containing the suspending agent is 1:1-1000 (v/v).

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

Said suspending agent comprises polyvinyl alcohol, sodium carboxymethyl cellulose (CMC-Na), polyvinylpyrrolidone (PVP) or a kind of natural colloid etc., and natural colloid comprises gelatin, acacia gum, pectin etc.

The method of using the preparation of the present invention is as follows: inject the sterile aqueous solution containing the suspending agent into the prepared microsphere ampoule, after mixing and shaking, it can be injected subcutaneously or intramuscularly, and the dosage should be handled according to the amount indicated in the instructions of different pharmaceutical preparations. .

The drug in the microspheres can be effectively released in the body for more than two weeks, so the treatment of endometriosis, uterine fibroids and other symptoms can be injected at least once at intervals of more than 15 days. The specific dose and interval of medication can be based on the specific situation of the patient Decide.

The drug encapsulated in the microsphere can be released under control 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 is absorbed through the peripheral blood circulation, thereby reducing the number of medications for the patient, which not only facilitates the treatment of the patient, but also reduces adverse reactions such as liver and kidney damage that may be caused by oral administration of the tablet.

Detailed ways

Example 1

Get 20mg gestrinone, 80mg polylactide-glycolide (lactide:glycolide=75:25, weight ratio), dissolve in 3ml methylene chloride, under vigorous stirring (1000rpm) Drop into 100ml 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, and filter the precipitate under reduced pressure , washed with a small amount of distilled water, dried under reduced pressure at room temperature, and the diameter of the microspheres is 40-120 μm.

Example 2

Take 20mg gestrinone and 80mgDL-polylactic acid, dissolve them in 1ml dichloromethane, add 5ml 0.5% PVA aqueous solution to a high-speed homogenizer, stir vigorously at 6000rpm for 2min, pour into 100ml 0.5% PVA aqueous solution, 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

Take 20mg nomegestone, 80mgDL-polylactic acid, dissolve in 1.25ml ethyl acetate, add 10ml 0.5% PVA aqueous solution in a high-speed homogenizer, stir vigorously at 8000rpm for 2min, pour into 100ml 0.5% PVA aqueous solution , evaporated at 300rpm at 25°C for about 12h, centrifuged, poured off the supernatant, filtered the precipitate under reduced pressure, rinsed the precipitate with a small amount of distilled water, dried under reduced pressure at room temperature, and the diameter of the microspheres was 20-40 μm.

Example 4

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 embodiment 2: the microsphere that single dose s.c. injection release rate is 0.5mg/week/kg, microsphere gestrinone content 25% (n=5), adopt 0.5% CMC-Na aqueous solution as auxiliary Suspension.

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 egg garden cells and keratinocytes. Explain that its estrous cycle has been continuously suppressed for 15 days. The negative control group was not suppressed.

Example 5

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 content at the transplantation site were those who successfully established the model. After the volume was measured, the animals were randomly divided into groups for the following treatment:

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 Example 2: Rats were injected with s.c. microspheres once at the back, the dose was 0.5 mg/week/kg, and the content of gestrinone in the microspheres was 25% (divided into 10 groups, n=3 in each group).

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. The results of the inhibition test of gestrinone microsphere injection on endometriosis in rats

Claims (12)

1, a kind of progestogen injection implants is characterized in that, comprises treatment effective amount of actives progestogen and as the biological degradation polyalcohol of framework material.

2, progestin preparation according to claim 1 is characterized in that, the weight percentage of progestogen is 1～70%.

3, progestin preparation according to claim 1 is characterized in that, said preparation is a microgranule, microsphere, a kind of in nanoparticle or the nanosphere.

4, progestin preparation according to claim 3 is characterized in that, microsphere is that diameter is 10～120 microns a circular entity.

5, progestin preparation according to claim 1, it is characterized in that said progestogen comprise a kind of in gestrinone, gestodene, nomegestrol, promegestone, norethindrone, megestrol, norgestrel, levonorgestrel, Desogestrel, norgestimate, Norethisteroneoxime, quingestrone or the anorethindrane dipropionate.

6, long-acting progestogen injection implants according to claim 1, it is characterized in that, said biological degradation polyalcohol comprises polylactide-co-glycolide, polylactic acid, polyglycolic acid, polycaprolactone, poly-anhydride, poly butyric ester-hydroxyl pentanoate copolymer, polypropylene glucosan, one or more of polylactic acid-polyglycol.

7, progestin preparation according to claim 6 is characterized in that, polymer molecular weight is 5,000～1,000,000, wherein, the part by weight of polylactide-co-glycolide monomer polymerization is 95: 5～5: 95, and molecular weight is 10,000-500,000, the polylactic acid molecule amount is 5,000-300,000.

8, a kind ofly contain the pharmaceutical composition that each described preparation of claim 1～7 for the treatment of effective dose and good biocompatibility and degradable carrier constitute.

9, pharmaceutical composition according to claim 8 is characterized in that, this pharmaceutical composition is subcutaneous or intramuscular dose or subcutaneous implant.

10, according to the preparation method of each described injectable progestogen microsphere of claim 1～9, it is characterized in that comprising the steps: progestogen and biological degradation polyalcohol are dissolved in the organic solvent, pour in the aqueous solution that contains suspending agent that is stirring, make O/w emulsion, fling to organic solvent, from this suspension, adopt conventional method to collect microsphere then, be preparation of the present invention.

11, preparation method according to claim 10 is characterized in that, degradable polymer content in organic solvent is 1～1000mg/ml;

Said organic solvent comprises dichloromethane, chloroform, ethyl acetate, acetonitrile or one or more;

Said suspending agent comprises polyvinyl alcohol, sodium carboxymethyl cellulose (CMC-Na), and polyvinylpyrrolidone (PVP), eucolloid, as gelatin, arabic gum, pectin etc. a kind of, the content of suspending agent in aqueous solution is 1～100mg/ml;

The organic solvent that contains progestogen and biological degradation polyalcohol is 1: 1～1000 (v/v) with the volume ratio that contains the aqueous solution of suspending agent.

12, according to of the application of each described progestin preparation of claim 1～9 in treatment endometriosis or hysteromyoma symptom.