WO2007009383A1

WO2007009383A1 - A rectum suppository of protein and its preparation method and the use

Info

Publication number: WO2007009383A1
Application number: PCT/CN2006/001755
Authority: WO
Inventors: Lei Jin; Jin Pei; Minghua Duan; Yuhui Zheng
Original assignee: Genescience Pharmaceuticals Co., Ltd.
Priority date: 2005-07-19
Filing date: 2006-07-19
Publication date: 2007-01-25
Also published as: CN1899604A

Abstract

A rectum suppository consisting of protein, suppository matrix, absorption enhancer, bacterial inhibitor, protective agent, pH regulator and/or antioxidant and its preparation method and the use are provided. Local administration (via rectum) may obtain systemic therapeutic effect. The suppository of the invention is convenient to use in clinic, metabolic process is long, and the therapeutic effect is perfect.

Description

Protein drug rectal suppository, preparation method and application thereof

The present invention relates to a protein drug rectal suppository, and more particularly to a protein drug rectal suppository which achieves systemic therapeutic action by local administration of the rectum, and a preparation method and application thereof.

Background technique

With the rapid development of bioengineering technology, biotechnology active drugs have been continuously introduced, and many biological products have been widely used in the prevention, treatment and prognosis of clinical diseases. According to reports, so far, more than 40 kinds of biological products have been approved for caps at home and abroad, and biologic drugs in the research and development stage have become hot spots in the pharmaceutical field.

Most of the bioactive drugs are protein drugs. Since the physicochemical properties of protein drugs determine the instability of their biological activities, the main dosage forms currently used clinically for protein drug preparations that require systemic administration are still liquid injections or lyophilized injections. The limitations of clinical application of such injections are mainly reflected in two aspects: The first aspect is reflected in the limitations of drug metabolism. After intravenous or subcutaneous injection, the drug has a short peak time and a high peak plasma concentration. It will soon be hydrolyzed by proteases in plasma, so the half-life of drug metabolism is short, which reduces its clinical therapeutic index. The second aspect is that for clinical patients, it is often necessary to receive daily injections, and the clinical use period is longer, long-term. Frequent injections cause a lot of pain to the patient, greatly reducing patient compliance. At present, the research and development of new dosage forms of protein drugs that achieve systemic therapeutic effects through non-injection drug delivery will become an urgent problem for many pharmaceutical companies and research institutes at home and abroad, inventing a solution that not only protects the active ingredient activity, It is also necessary to use a new dosage form that is convenient for clinical use and has obvious effects.

The development of new formulations of protein drugs mainly includes long-acting, controlled-release, oral, nasal spray, suppository, etc., and achieved certain results. The protein drug suppositories marketed at home and abroad all achieve the purpose of local action by topical administration. Such as interferon ot-2b suppository clinical indication for the treatment of viral infection caused by (or at the same time) cervical erosion; interferon a-2a suppository clinical indication for the treatment of vaginal viral infection caused by chronic cervicitis, cervical erosion, Vaginitis, prevention of cervical cancer. To date, there have been no reports of protein drug suppository dosage forms that achieve systemic therapeutic effects by topical administration at home and abroad. When protein and peptide drugs are administered through the rectum, the main problems they face are poor biofilm permeability and loss of activity. Once the protein drug passes through the rectal mucosa, it can enter the liver through the upper rectal vein and then run through the whole body or through the middle and lower rectal veins and anal canal veins, bypassing the liver, and directly entering the blood circulation from the inferior vena cava. At the same time, the drug can also be absorbed by the rectal mucosa through the rectal mucosa and enter the lymphatic system. Therefore, the rectal lymphatic system absorbs the drug almost in the same position as the blood. Therefore, inventing a high-absorbency and stable protein drug suppository has become the key to the research and development of this dosage form.

Summary of the invention

SUMMARY OF THE INVENTION The object of the present invention is to provide a protein drug rectal suppository which achieves systemic therapeutic action by local administration of the rectum.

Another object of the present invention is to provide a method of preparing the above-described protein drug suppository. It is also an object of the present invention to provide the use of the above-described protein drug suppositories. .

In order to achieve the above object, the present invention provides a protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum, and is composed of the following components: a protein drug, a suppository base, a protein protectant, an absorption enhancer, a bacteriostatic agent. , pH adjusters and / or antioxidants. The components are in the percentage by weight: protein drug 0.00001~5%, suppository matrix 60~99%, absorption enhancer 0.001~10%, bacteriostatic agent>0~5%, protein protectant 0.00001~10%, pH Conditioner > 0 to 30% and / or antioxidant > 0 ~ 5%.

The components are preferably 0.001 to 5% by weight of the protein, 60 to 90% of the suppository base, 0.001 to 10% of the absorption enhancer, 0.001 to 5% of the bacteriostatic agent, 0.001 to 10% of the protein protective agent, and pH adjustment. Agents > 0 to 30% and / or antioxidants > 0 ~ 5%.

The proteinaceous drug is preferably recombinant human growth hormone.

The proteinaceous drug is preferably an interferon.

The proteinaceous drug is also preferably one or more of the following components: insulin, thymosin al, recombinant human granulocyte colony stimulating factor, somatostatin, glucose oxidase, heparin, superoxide dismutase, tissue Growth factors, recombinant human granulocyte macrophage stimulating factor, follicle stimulating hormone, erythropoietin, tissue growth factor, coagulation factor and/or cytokine.

The absorption enhancer is preferably one or more of the following components: laurel, formyldimethylamine, dimethyl sulfoxide, ethyl nicotinate, propylene glycol, aspartame, nonoxynol Ether, hesperidin, poloxamer, trapolone, vitamin E succinate and/or neohesperidin dihydrochalcone. The suppository base is a water soluble suppository base or a fat soluble suppository base.

The water-soluble suppository base is preferably one or more of the following components: glycerin gelatin, polyethylene glycol, polyoxyethylene stearate, Tween-61, Tween-60, Tween-80, Tween-20 and / or carbomer.

The fat-soluble suppository base is preferably one or more of the following components: cocoa butter, fragrant fruit fat, semi-synthetic cocoate, mixed fatty acid glyceride, semi-synthetic fatty acid ester, propylene glycol stearate, black glutinous rice Fat, fractionated palm oil, fractionated coconut oil, carnauba wax, glyceryl tripalmitate, glyceryl tristearate, hydrogenated vegetable oil, cocambide and/or semi-synthetic bacteriostatic agent are preferably the following ingredients One or more of: sorbic acid, phenol, cresol, chlorocresol, propionic acid, benzoic acid, dehydroacetic acid, methylparaben, ethylparaben and/or propylparaben.

The protein protectant is preferably one or more of the following components: trehalose, sucrose, mannitol, histidine, glycine, methionine, sodium tartrate, sodium succinate, cyclodextrin, ethylenediaminetetraacetic acid Disodium, human albumin, gelatin, sodium chloride and/or dextran.

The pH adjusting agent is preferably one or more of the following components: hydrochloric acid, sodium hydroxide, sodium carbonate, sodium hydrogencarbonate, citric acid, acetic acid, sodium acetate, maleic acid, sodium maleate, cesium Sodium citrate, boric acid, sodium borate, citric acid and/or sodium citrate.

The antioxidant is preferably one or more of the following components: Nibble methyl ester, Nipagin ethyl ester, Nipa gold propyl ester, sorbic acid, benzoic acid and salts, tartaric acid and salts, Trichlorotert-butanol, thimerosal, ascorbic acid, tocopherol and/or sodium thiosulfate.

The present invention also provides a method for preparing a protein rectal suppository which achieves systemic therapeutic effect by local administration of the rectum. When the suppository base is a water-soluble suppository base, the following steps are included: the bacteriostatic agent, the absorption enhancer, the antioxidant and the pH adjuster are mixed and heated according to a certain proportion of the suppository substrate, and cooled to 35-60 ° C to add the protein drug And protein protectants, filling, cooling, forming, demoulding, packaging, that is.

When the suppository base is a fat-soluble suppository base, the method comprises the steps of: first melting the fat-soluble matrix at 65 75 ° C, and respectively adding an absorbent, an antioxidant, a bacteriostatic agent and a pH adjuster to the molten matrix, After mixing evenly, the temperature is lowered to 35~60 °C, adding protein drugs and protein protection to cut, mold, cool, form, demould, and package.

The invention provides a recombinant human growth hormone rectal suppository which achieves systemic therapeutic effect by local administration of the rectum in the preparation of a hypothalamic-pituitary for treating children with slow growth, severe burns and phlegm caused by endogenous growth hormone deficiency. Disease-induced growth hormone Deficiency, a significant lack of growth hormone confirmed by two different growth hormone tests, a negative nitrogen balance after surgery, and an anti-aging drug.

The present invention provides an interferon rectal suppository which achieves systemic therapeutic effect by local administration of the rectum for the preparation of a medicament for the treatment of chronic hepatitis B, chronic hepatitis C, condyloma acuminata, cytoskeletal leukemia and chronic myeloid leukemia.

The protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to the invention has convenient clinical use, long drug metabolism time and obvious therapeutic effect.

The essence of the present invention will be exemplified by the results of the stability test of the recombinant human growth hormone plug and the recombinant human interferon a-2b plug and the results of the absorbance test in the animal body.

1. Recombinant human growth hormone suppository stability test

The stability of the recombinant human growth hormone-deficient suppository of the present invention for long-term preservation at 4 °C was examined. The results showed that the recombinant human growth hormone plug can be stored for 2 years at 4 °C. The experimental results show that according to the requirements of the Chinese Pharmacopoeia on the stability study of recombinant human growth hormone, the liquid phase determination of the relevant proteins and high molecular proteins of the three batches of samples are in line with the national pharmacopoeia standards, which proves the biological activity of recombinant human growth hormone suppositories. stability. The specific results are shown in Tables 1 and 2.

Table 1 Recombinant human growth hormone plug 4Γ stability test results (RP-HPLC)

Table 2 Recombinant human growth hormone plug 4 ° C stability test results (SEC-HPLC)

2, interferon a-2b suppository stability test

The stability of the interferon a-2b suppository for treating hepatitis and herpes virus of the present invention for long-term storage at 4 ° C was examined. The results showed that the interferon ot-2b plug can be stored for 2 years at 4 ° C. The results of the experiment indicated that: according to the requirements of the Chinese Pharmacopoeia on the stability study of interferon o-2b, the biological activity of interferon a-2b suppository was determined by cell virus inhibition method. The results of the three batches of samples were in line with the national pharmacopoeia standards. , proved interferon a-2b The stability of the biological activity of suppositories. The specific results are shown in Table 3.

Table 3 Interferon cc-2b plug 4 Stability test results

3. Resorptive test of recombinant human growth hormone in rabbits

Test objective: To observe the changes in the activity of recombinant human growth hormone in suppositories and their absorption in animals.

experiment method

Source of Drugs: Recombinant human growth hormone is derived from Changchun Jinsai Pharmaceutical Co., Ltd. 巿 Shangyu products, the trade name is Sai Zeng.

Excipient blank plug: Semi-synthetic fatty acid glyceride blank plug.

Test article: Recombinant human growth hormone thrombus.

Control substance: recombinant human growth hormone freeze-dried powder needle.

Animals: Rabbits, weighing 2 kg or so, male and female. Provided by the Animals Department of Tianjin Pharmaceutical Research Institute.

Animal grouping: Three groups (excipient blank plug group, recombinant human growth hormone freeze-dried powder and recombinant human growth hormone plug group), three in each group.

Dosage: Per rabbit by weight: 33.33mg/kg

Method of administration: At 25±2°C and 65% humidity, the rabbits were divided into 3 groups, 3 in each group, half male and half female, fasting for 18 hours (not allowed for water). The rabbits were first fixed in a rabbit fixation frame. After lh, blood was taken from the ear vein for 50 μl and then administered. Each group of suppositories. · Embryo blank plug and recombinant human growth hormone were inserted into the anus 2cm of the rabbit, and the anus was ligated with a wire rope, once a time, once a day, and administered rectally. 2 Subcutaneous injection of recombinant human growth hormone group: Recombinant human growth hormone solution (4.5 IU/ml) was injected into rabbit hip muscles by rabbit body weight (0.2 ml/kg) (0.075 IU/ml) once a day.

Blood collection method: Each group was continuously collected for blood at a prescribed time, 50μ each time, after centrifugation, the upper layer of plasma was taken, and the recombinant human growth hormone content was determined by radioimmunoassay.

test results

Preparation of standard curve

Precisely measure the blank plasma sample 100μ into the recombinant human growth hormone series standard solution, so that the concentration of recombinant human growth hormone in plasma is: 0.5, 1.0, 2.0, 4.0, 8.0 Ng/mL. The standard curve was drawn by taking the different concentrations of standard recombinant human growth hormone as the abscissa and the corresponding radioactivity obtained in the measurement as the ordinate. The curve equation is: y=0.0271x ² - 0.3119X + 0.8573.

Recovery rate determination

A certain amount of recombinant human growth hormone solution was accurately added to the blank plasma, and the results were analyzed by radioimmunoassay. The absolute recovery was calculated by comparing with the respective concentrations. The results are shown in Table 4.

Table 4 Recovery rate test results

The results showed that the average recoveries of recombinant human growth hormone in the plasma at low, medium and high levels were 90.00%, 92.831%, 89.96%, respectively. The average relative standard deviation was 4.44%, 0.16%, and 0.65%.

Precision measurement

A certain amount of recombinant human growth hormone solution was accurately added to the blank plasma, and the determination result was analyzed by radioimmunoassay. The same sample solution was measured 5 times a day to calculate the relative standard deviation within the sputum; the same batch of samples was measured 5 times within 5 days, and the daytime relative was calculated. Standard deviation, the results are shown in Table 5.

Table 5 precision experiment results

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Growth hormone concentration average RSD concentration average RSD (ng/ml) (ng/ml) (%) (ng/ml) {%)

0.5 0.44 1.75 0.43 0.15

2.0 1.82 0.85 1.84 0.84 0 7.18 0.25 7.20 0.72 The results showed that the intra-day average relative standard deviation of recombinant human growth hormone in plasma at low, medium and high levels was 1.75%, 0.85%, 0.25%, and recombinant human growth hormone was low, medium and high in plasma. The average relative standard deviations at daytime concentrations were 0.15%, 0.84%, and 0.72%, as shown in Tables 6, 7, and 8.

Table 6 Results of subcutaneous injection of recombinant human growth hormone lyophilized powder

Time, recombinant human growth hormone, blood concentration, blood collection point

( hr ) ( g/ml ) ±SD

1 0.25 0.86 ± 0.59

2 0.5 1.50±0.42

3 1.0 3.80±1.12

4 1.5 6.22±0.79

5 2.0 5.61±1.05

6 3.0 4.27±0.13

7 4.0 3.24±0.46

8 8.0 2.18±1.18

9 12.0 1.13±0.48

10 24.0 0.82±0.43

Table 7 blank plug measurement results

( hr ) ( ng/ml ) ±SD

1 0.25 0.00±0.00

2 0.5 0.00±0.00

3 1.0 0.00±0.00

4 1.5 0.00±0.00

5 2.0 0.00±0.00

6 3.0 0.00±0.00

7 4.0 0.00±0.00

8 8.0 0.00+0.00

9 12.0 0.00±0.00

10 24.0 0.00+0.00

Recombinant human growth hormone suppository assay results Time recombination human growth hormone blood concentration blood point

( hr ) ( g ml ) +SD

1 0.25 0.84 ± 0.27

2 0.5 0.89±1.17

3 1.0 2.41±0.37

4 1.5 4.25±0.34

5 2.0 5.72+0.27

6 3.0 4.53±0.32

7 4.0 3.15+0.20

8 8.0 2.43±0.68

9 12.0 1.24±0.97

10 24.0 0.91±1.26

From the above results, it can be seen that the blank plug has no effect on the detection of recombinant human growth hormone in the blood. The relative bioavailability of the recombinant human growth hormone suppository in rabbits is 92.57%, which is bioequivalent to the recombinant human growth hormone subcutaneous injection. It is indicated that recombinant human growth hormone thrombus can achieve systemic therapeutic effects through anal administration.

4. Efficacy test of recombinant human growth hormone thrombus in pituitary rats

OBJECTIVE: To determine whether recombinant human growth hormone subcutaneous injection and three different doses of recombinant human growth hormone suppository have the effect of promoting growth and development of pituitary rats.

Test Methods 'Clean-grade Wistar rats were used to remove the pituitary gland. The effects of recombinant human growth hormone subcutaneous injection and three different doses of recombinant human growth hormone suppository on the growth and development of pituitary gland were observed.

Source of Drugs: Recombinant human growth hormone is derived from Changchun Jinsai Pharmaceutical Co., Ltd., which has been listed as a product.

Excipient blank plug: Semi-synthetic fatty acid glyceride blank plug.

Test article: Recombinant human growth hormone thrombus.

Control substance: recombinant human growth hormone freeze-dried powder needle.

Animals: Pituitary rats, weighing 80.4 ± 4.1 g, male. Provided by the Animal Experimental Center of the Basic Medical College of Jilin University.

Method of administration: The temperature of 25 ± 2 ° C, humidity 65%, the pituitary rats were divided into 5 groups, 8 rats in each group, male, fasting for 18h (can not help water). Then administered, once a day. test results

Recombinant human growth hormone injection has a significant effect on promoting the growth and development of pituitary rats; recombinant human growth hormone suppository has a significant effect on promoting the growth and development of pituitary rats, and in the dose range of 50 _μβ / only to 200 μ ^ The inside is linear with the dose. There were significant differences between the three suppository groups and the model group (Ρ<0.05). There was no significant difference between the suppository group and the injection group (Ρ>0.05). The injection group had significant difference compared with the model group. (Ρ<0.05). The measurement results are shown in Table 9.

Table 9

5, recombinant human interferon cx-2b suture in rabbits in vivo absorption test

Test purposes

The activity of recombinant human interferon o-2b in suppositories and its absorption in animals were observed.

experiment method

Source of drugs: Interferon a-2b is derived from Changchun Jinsai Pharmaceutical Co., Ltd. Test lot number: Recombinant human interferon a-2b suppository (20041101) by Changchun Jinsai drug Industry limited liability company

Excipients: Glycerin gelatin plug

Control substance: recombinant human interferon ot-2b freeze-dried powder needle (commercial products of Changchun Institute of Biological Products)

Animal grouping: A total of three groups (excipient blank plug group, subcutaneous injection of commercially available interferon a-2b freeze-dried powder group and recombinant human interferon a-2b suppository group), three in each group.

Dosage: Per rabbit by weight: 1.25 x ^{10 5} IU/kg.

Method of administration: At 25±2°C and 65% humidity, rabbits were divided into 3 groups, 3 in each group, half male and half female, fasting for 18 hours (not allowed for water). The rabbits were first fixed in a rabbit fixation frame, and after lh, 50 μl of blood was taken from the ear vein, and then administered. Each group of suppositories: The excipient blank plug and interferon cx-2b were inserted into the anus 2 cm of the rabbit, and the anus was ligated with a wire rope, once a time, once a day, and administered rectally. 2 Subcutaneous injection of interferon a-2b group: Interferon a-2b solution (300xl0 ⁴ IU/ml) was injected into rabbit buttock muscles by syringe (1.25xl0 ⁵ IU / kg) once a day.

Blood collection method: Each group was continuously collected for blood at a prescribed time, 50 L each time, after centrifugation, the upper layer of plasma was taken, and the interferon a-2b content was determined by enzyme-free method.

test results

Preparation of standard curve

The blank plasma sample lOOuL was accurately weighed and added to the interferon oc-2b series standard solution to make the plasma interferon- 2b concentration: 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0, 32.0 ng/mL. The standard drug concentration was plotted on the abscissa with the different drug concentrations of the standard interferon a-2b, and the corresponding OD values obtained in the assay were plotted on the ordinate. The results of the measurements are shown in Table 10.

Table 10

Drug concentration (ng mL) 0.25 0.5 1.0 2.0 4.0 8.0 16.0 32.0

OD value 2459 2438 2399 1832 1398 1248 1198 1200 Determination of recovery

A certain amount of interferon a-2b solution was accurately added to the blank plasma, and the results were analyzed by enzyme immunoassay. The absolute recovery was calculated by comparing with the respective concentrations. The results are shown in Table 11. Table 11 Recovery rate experiment results

The results showed that the average recoveries of interferon a-2b in the plasma at low, medium and high levels were 90.00%, 92.831%, 89.96%, respectively. The average relative standard deviation was 4.44%, 0.155%, and 0.65%.

Precision measurement

A certain amount of interferon a-2b solution was accurately added to the blank plasma, and the results of the enzyme assay were analyzed. The same sample solution was measured 5 times a day to calculate the relative standard deviation in the sputum; the same batch of samples was measured 5 times within 5 days to calculate 曰The relative standard deviation between the two, the results are shown in Table 12.

Table 12 precision test results

Into interferon

A-2b concentration

Concentration average (ng/ml) RSD (%) Concentration average (ng/ml) RSD (%) ( ng / ml )

0.5 0.44 1.75 0.43 0.15

2.0 1.82 0.85 1.84 0.84

8.0 7.18 0.25 7.20 0.72 The results showed that the average relative standard deviation of interferon a-2b in the plasma at low, medium and high levels of plasma was 1.75%, 0.85%, 0.25%, interferon a-2b in plasma. The average relative standard deviation of daytime at low, medium and high levels was 0.15%, 0.84%, 0.72%. In accordance with the Pharmacopoeia regulations, see Tables 13, 14, and 15.

Table 13 Interferon a-2b sale freeze-dried powder needle subcutaneous injection test results Time concentration

(hr) (ng/ml) ±SD

1 0.25 0.87±0.35

2 0.5 1.48+0.27

3 1.0 3.95±0.27

4 1.5 6.28±0.97

5 2.0 5.64±0.18

6 3.0 4.28±0.93

7 4.0 3.28±0.29

8 8.0 2.17±0.58

9 12.0 1.19±1.05

10 24.0 0.84±0.85 Table 14 Blank plug measurement results

Time concentration

(hr) (ngml) ±SD

1 0.25 0.00+0.00

2 0.5 O.OOlO.OO

3 1.0 0.00±0.00

4 1.5 O.OOlO.OO

5 2.0 0.00+0.00

6 3.0 O.OOlO.OO

7 4.0 0.00±0.00

8 8.0 O.OOlO.OO

9 12.0 O.OOlO.OO

10 24.0 0.00±0.00 Table 15 Interferon oc-2b suppository measurement time concentration blood collection point

(hr) ( ng/ml ) ±SD

1 0.25 0.83 ± 0.28

2 0.5 4.28±0.98

3 1.0 5.73±0.23

4 1.5 4.45±0.18

5 2.0 4.08+0.78 6 3.0 3.52±0.27

7 4.0 3.12±1.02

8 8.0 2.46+0.85

9 12.0 1.21±1.08

10 24.0 0.90±0.52

By fitting the above parameters, it can be seen that the blank plug has no effect on the detection of interferon oc-2b in the blood. The relative bioavailability of interferon o-2b suppository in rabbits is 97.3%, and interferon injection of interferon a-2b. Bioequivalent. Explain that interferon a-2b suppository can achieve systemic treatment through anal administration.

detailed description

The invention is further illustrated by the following examples, which are not intended to limit the invention.

Example 1

500 mg of semi-synthetic fatty acid glyceride was melted at 65 ° C, and 25 mg of trehalose, 25 mg of cyclodextrin, and 0.1 mg of ascorbic acid were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 35 ° C, and the recombinant life was added. Long hormone 17.0mg, filling mold, cooling, molding, demoulding, packaging.

Example 2

750 mg of semi-synthetic glyceride was melted at 75 ° C, and 30 mg of sucrose, 30 mg of glycine, 20 mg of methionine, Tween-80 200 μl, nonoxynol 50 mg, sodium sorbate 0.05 mg, and Nibopol methyl ester 0.05 m _g were respectively After being added to the molten matrix, the mixture was uniformly mixed, the temperature was lowered to 60 ° C, and recombinant human growth hormone was added to 17.0 mg, filled, cooled, formed, demolded, and packaged.

Example 3

100 mg of semi-synthetic fatty acid glyceride was melted at 70 ° C, and 50 mg of histidine, 60 mg of sucrose, 70 mg of laurel, 0.03 mg of sodium sorbate, 0.07 mg of nipagin ethyl ester, and 0.1 mg of sodium hydroxide. They were added to the molten matrix separately, and after mixing uniformly, the temperature was lowered to 40 ° C, and recombinant human growth hormone was added to 17.0 mg, filled, cooled, formed, demolded, and packaged.

Example 4

The semi-synthetic fatty acid glyceride 740m _g was melted at 68 ,, and sucrose 35 mg, glycine 35 mg, Tween-80300 μl, and nisparin propyl acetate were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50°. C, adding recombinant human growth hormone 17.0mg, irrigation Mold, cooling, forming, demoulding, packaging.

Example 5

640 mg of semi-synthetic fatty acid glyceride was melted at 72 ° C, 30 mg of trehalose, 30 mg of cyclodextrin, and 0.02 mg of nipagin ethyl ester were added to the molten matrix, and after mixing, the temperature was lowered to 55 Ό, and the mixture was added. Human Growth Hormone 17.0mg, Filling, Cooling, Molding, Demoulding, Packaging.

Example 6

550 mg of semi-synthetic fatty acid glyceride was melted at 73 ° C, and 25 mg of trehalose, 20 mg of cyclodextrin, and 0.03 mg of sodium sorbate were separately added to the molten matrix. After mixing, the temperature was lowered to 50 ° C, and the mixture was added. Human Growth Hormone 17.0mg, Filling, Cooling, Molding, Demoulding, Packaging.

Example 7

750 mg of semi-synthetic fatty acid glyceride was melted at 67 ° C, 40 mg of trehalose, 40 mg of cyclodextrin, and 0.04 mg of nipagin ethyl ester were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50 ° C. Recombinant human growth hormone 37.0 mg was added, filled, cooled, shaped, demolded, packaged.

Example 8

500mg cocoa butter melted at 70 ° C, trehalose 20mg, cyclodextrin 20m _g, ascorbic acid O.Olmg, are added to the molten matrix, after mixing, temperature was lowered to 50 ° C, recombinant human growth hormone 27.0mg, filling, cooling, forming, demoulding, packaging.

Example 9

550mg of cocoa butter was melted at 70 ° C, 30mg of trehalose, 30mg of cyclodextrin, O.Omg of tocopherol, respectively, were added to the molten matrix, and after mixing, the temperature was lowered to 50 ° ( Adding recombinant human growth hormone 25.0mg, filling, cooling, forming, demoulding,

package.

Example 10

100 mg of cocoa butter was melted at 70 ° C, and 50 mg of trehalose, 50 mg of cyclodextrin, and O.Olmg of nipagin were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50 ° C, and the mixture was added. Human Growth Hormone 34.0mg, Filling, Cooling, Forming, Demoulding, Packaging.

Example 11

Glycerin gelatin 500mg, trehalose 20mg, cyclodextrin 30mg, ascorbic acid O.Olmg, after heating and mixing at 65 ° C, the temperature was lowered to 35 ° C, and recombinant human growth hormone 7.0 mg was added, filled, cooled, formed, demolded, and packaged.

Example 12

750mg of polyoxyethylene ( 40 ) stearate, 20mg of trehalose, 60mg of cyclodextrin, 0.02mg of Nibium ethyl ester, heated and mixed at 70 ° C, the temperature dropped to 60 ° C, added recombinant human growth Hormone 17.0mg, filling, cooling, molding, demoulding, packaging.

Example 13

1000mg of polyethylene glycol, 50mg of histidine, 60mg of cyclodextrin, 0.04mg of Nibopol methyl ester, heated and mixed at 75 ° C, the temperature was reduced to 50 ° C, and recombinant human growth hormone was added to 17.0 mg. Mold, cooling, forming, demoulding, packaging.

Example 14

450mg of glycerin gelatin, 20mg of trehalose, 30mg of glycine, O.Olmg of propylglycolate, heated and mixed at 68 ° C, the temperature was lowered to 40 ° C, 15.0 mg of recombinant human growth hormone was added, and the mold was poured and cooled. Molding, demoulding, packaging.

Example 15

650 mg of polyethylene glycol, 30 mg of trehalose, 30 mg of cyclodextrin, and O.Olmg of Nibopol ethyl ester were heated and mixed at 72 ° C, the temperature was lowered to 5 CTC, and 20.0 mg of recombinant human growth hormone was added to form a mold. Cooling, forming, demoulding, packaging.

Example 16

750mg of polyoxyethylene ( 40 ) stearate, 30mg of trehalose, 50mg of cyclodextrin, 0.05mg of sodium sorbate, heated and mixed at 70 ° C, the temperature was lowered to 55 ° C, and recombinant human growth hormone was added. Mg, filling, cooling, forming, demoulding, packaging.

Example 17

750mg of glycerin gelatin, 35mg of trehalose, 50mg of cyclodextrin, 0.06mg of nipagin ethyl ester, heated and mixed at 70 °C, the temperature was lowered to 50 ° C, 30.0 mg of recombinant human growth hormone was added, and the mold was cooled. , molding, demoulding, packaging.

Example 18

Polyoxyethylene (40) stearate 500mg, trehalose 25mg, cyclodextrins _g, ascorbic acid O.Olmg 25m, was heated at 70 ° C after mixing, temperature was lowered to 45 ° C, recombinant human growth hormone was added 25.0 Mg, filling, cooling, forming, demoulding, packaging.

Example 19

700mg of glycerin gelatin, 30mg of trehalose, 30mg of cyclodextrin, O.Omg of tocopherol, heated and mixed at 70 ° C, the temperature was lowered to 50 ° C, and recombinant human growth hormone was added. 34.0mg, filling, cooling, forming, demoulding, packaging.

Example 20

1000mg of polyethylene glycol, 50mg of trehalose, 50mg of cyclodextrin, O.Olmg of Nibopol ethyl ester, heated and mixed at 70 °C, the temperature was lowered to 40 ° C, and recombinant human growth hormone was added to 34.0 mg. Mold, cooling, forming, demoulding, packaging.

Example 21

480 mg of semi-synthetic fatty acid glyceride was melted at 70 ° C, and 25 mg of trehalose, 25 mg of cyclodextrin, and 0.1 mg of ascorbic acid were separately added to the molten matrix. After mixing uniformly, the temperature was lowered to 50 ° C, and interferon was added. _a -2b 50xl0 ⁷ IU (equivalent to 5.0mg), filling, cooling, forming, demoulding, packaging.

Example 22

650 mg of semi-synthetic fatty acid glyceride was melted at 70 ° C, and 30 mg of trehalose, 40 mg of cyclodextrin, and O.Olmg of Nibethyl ethyl ester were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50 ° C. , add interferon a-2b 30xl0 ⁷ IU (equivalent to 3.0mg), filling, cooling, molding, demoulding, packaging.

Example 23

720 mg of semi-synthetic fatty acid glyceride was melted at 70 ° C, and 35 mg of histidine, 45 mg of cyclodextrin, and 0.1 mg of Nibopol methyl ester were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50 °. C, add interferon a-2b 5.0xl0 ⁷ IU (equivalent to 0.50mg), mold filling, cooling, molding, demoulding, packaging.

Example 24

675 mg of semi-synthetic fatty acid glyceride was melted at 70 ° C, and 30 mg of trehalose, 45 mg of glycine, and 1.0 mg of propyl propyl methoxide were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50 ° C, and added. Interferon a-2b 1.5xl0 ⁷ IU (equivalent to 0.15mg), filling, cooling, forming, demoulding, packaging.

Example 25

747 mg of semi-synthetic fatty acid glyceride was melted at 70 ° C, and 25 mg of trehalose, 58 mg of cyclodextrin, and O.Olmg of Nibopol were added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50 ° C. , add interferon a-2b 15xl0 ⁷ IU (equivalent to 1.5mg), filling, cooling, molding, demoulding, packaging.

Example 26

630 mg of semi-synthetic fatty acid glyceride was melted at 70 ° C, and 35 mg of trehalose, 35 mg of cyclodextrin, and 0.1 mg of sodium sorbate were respectively added to the molten matrix, and after mixing, The temperature was lowered to 50 ° C, and interferon a-2b 20 x ^{10 7} IU (equivalent to 2.0 mg) was added, filling, cooling, molding, demoulding, and packaging.

Example 27

450 mg of semi-synthetic fatty acid glyceride was melted at 70 Torr, 25 mg of trehalose, 25 mg of cyclodextrin, and 0.1 mg of Nibopol ethyl ester were respectively added to the molten matrix, and after mixing uniformly, the temperature was lowered to 50 ° C, and added. Interferon a-2b 25xl0 ⁷ IU (equivalent to 2.5mg), filling, cooling, forming, demoulding, packaging.

Example 28

360 mg of cocoa butter was melted at 70 ° C, and 20 mg of trehalose, 20 mg of cyclodextrin, and 0.1 mg of ascorbic acid were respectively added to the molten matrix. After mixing uniformly, the temperature was lowered to 50 ° C, and interferon a-2b was added. 30xl0 ⁷ IU (equivalent to 3.0mg), filling, cooling, forming, demoulding, packaging.

Example 29

720 mg of cocoa butter was melted at 70 ° C, and 40 mg of trehalose, 40 mg of cyclodextrin, and 0.1 mg of tocopherol were respectively added to the molten matrix. After mixing uniformly, the temperature was lowered to 5 CTC, and interferon a-2b 35xl0 was added. ⁷ IU (equivalent to 3.5mg), filling, cooling, forming, demoulding, packaging.

Example 30

900 mg of cocoa butter was melted at 70 ° C, and 50 mg of trehalose, 50 mg of cyclodextrin, and O.Olmg of nipagin were respectively added to the molten matrix. After mixing uniformly, the temperature was lowered to 50 ° C, and interference was added. A-2b 45xl0 ⁷ IU (equivalent to 4.5mg), filling, cooling, forming, demoulding, packaging.

Example 31

450mg of glycerin gelatin, 30mg of trehalose, 20mg of cyclodextrin, O.Omg of ascorbic acid, heated and mixed at 70 ° C, the temperature was reduced to 50 ° C, interferon a-2b 50xl7 ⁷ IU (equivalent to 5.0mg) , filling, cooling, forming, demoulding, packaging.

Example 32

Polyoxyethylene (40) stearate 630mg, trehalose 30mg, cyclodextrin 40m _g, Winnipeg gold ethyl O.Olmg, heated at 70 ° C after mixing, temperature was lowered to 50 ° C, was added interference A- 2b 65xl0 ⁷ IU (equivalent to 6.5mg), filling, cooling, forming, demoulding, packaging.

Example 33

585mg polyethylene glycol, 30mg histidine, 35mg cyclodextrin, methyl nibble O.Olmg, after heating and mixing at 70 ° C, the temperature was lowered to 50 ° C, interferon a-2b 70xl0 ⁷ IU (equivalent to 7.0mg), filling, cooling, molding, demoulding, packaging.

Example 34

495 mg of glycerin gelatin, 20 mg of trehalose, 35 mg of glycine, and 100 mg of propyl propyl methoxide were heated and mixed at 70 ° C, and the temperature was lowered to 50 Ό, and interferon a-2b 75× ^{10 7} IU (equivalent to 7.5 m _{g) was added.} ), filling, cooling, forming, demoulding, packaging.

Example 35

540mg of polyethylene glycol, 30mg of trehalose, 30mg of cyclodextrin, O.Olmg of Nibopol ethyl ester, heated and mixed at 70 ° C, the temperature was lowered to 50 ° C, and interferon a-2b 1.5xl0 ^{7 was added.} IU (equivalent to 0.15mg), filling, cooling, forming, demoulding, packaging.

Example 36

540mg of polyoxyethylene ( 40 ) stearate, 20mg of trehalose, 40mg of cyclodextrin, O.Olmg of sodium sorbate, heated and mixed at 70 ° C, the temperature was lowered to 50 ° C, and interferon a- was added. 2b 1.5xl0 ⁷ IU (equivalent to 0.15mg), filling, cooling, forming, demoulding, packaging.

Example 37

495 mg of glycerin gelatin, 35 mg of trehalose, 20 mg of cyclodextrin, and O.Olmg of Nipagin ethyl ester were heated and mixed at 70 ° C, and the temperature was lowered to 50 ° C, and interferon a-2b 1.5× ^{10 7} IU was added. Equivalent to 0.15m _g ), filling, cooling, forming, demoulding, packaging.

Example 38

540mg of polyoxyethylene ( 40 ) stearate, 30mg of trehalose, 30m _{g of} cyclodextrin, O.Omg of ascorbic acid, heated and mixed at 70 ° C, the temperature was lowered to 50 ° C, and interferon a-2b was added. 1.5xl0 ⁷ IU (equivalent to 0.15mg), filling, cooling, forming, demoulding, packaging.

Example 39

Glycerin gelatin 630mg, trehalose 30mg, cyclodextrin 40mg, tocopherol 0.01mg, heated and mixed at 70 ° C, the temperature dropped to 50 ° C, added interferon a-2b 1.5xl0 ⁷ IU (equivalent to 0.15mg ), filling, cooling, forming, demoulding, packaging.

Example 40

900mg of polyethylene glycol, 50mg of trehalose, 50mg of cyclodextrin, O.Olmg of Nibopol ethyl ester, heated and mixed at 70 ° C, the temperature was lowered to 50 ° C, and interferon a-2b 3.0xl0 ^{7 was added.} IU (equivalent to 0.3mg), filling, cooling, forming, demoulding, packaging.

Claims

1. A protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum, and is characterized by the following components: a protein drug, a suppository base, an absorption enhancer, a bacteriostatic agent, a protein protectant, a pH adjuster And / or antioxidants.

2. A protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to claim 1, wherein each component is in a percentage by weight: a protein drug 0.00001 to 5%, a suppository substrate 60~ 99%, absorption enhancer 0.001-10%, bacteriostatic agent >0~5%, protein protectant 0.00001-10%, pH adjuster >0~30% and/or antioxidant >0~5%.

3. A protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to claim 2, wherein each component is in the range of weight percent: protein drug 0.001 to 5%, suppository matrix 60 ~ 90%, absorption enhancer 0.001 ~ 10%, bacteriostatic agent > 0 ~ 5 %, protein protectant 0.001 ~ 10%, pH regulator > 0 ~ 30% and / or antioxidant > 0 ~ 5 %.

4. A protein drug rectal suppository which achieves systemic therapeutic action by local administration of the rectum according to any one of claims 1 to 3, wherein the protein drug is recombinant human growth hormone.

5. A protein drug rectal suppository which achieves systemic therapeutic action by local administration of the rectum according to any one of claims 1 to 3, wherein the protein drug is recombinant human interferon.

6. A protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to any one of claims 1 to 3, wherein the protein drug is one or more of the following components. Species: insulin, thymosin al, recombinant human granulocyte colony-stimulating factor, somatostatin, glucose oxidase, heparin, superoxide dismutase, tissue growth factor, recombinant human granulocyte macrophage stimulating factor, follicle stimulating hormone , erythropoietin, tissue growth factor, coagulation factor and/or cytokine.

7. A protein drug rectal suppository according to any one of claims 1 to 3, wherein the absorption enhancer is one or more of the following components. Species: Lauryl azide, formyldimethylamine, dimethyl sulfoxide, ethyl nicotinate, propylene glycol, aspartame, nonoxynol, hesperidin, poloxamer, travazone, vitamins E succinate and / or neo-hesperidin dihydrochalcone.

8. A method according to any one of claims 1 to 3, which achieves the whole body by local administration of the rectum The therapeutic protein rectal suppository is characterized in that, when the suppository base is a water-soluble suppository base, it is one or more of the following components: glycerin gelatin, polyethylene glycol, polyoxyethylene stearate , Tween-61, Tween-60, Tween-80, Tween-20 and/or Carbomer.

The protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to any one of claims 1 to 3, wherein the suppository base is a fat-soluble suppository base, which is the following component One or more of: cocoa butter, fragrant fruit fat, semi-synthetic cocoate, mixed fatty acid glycerides, semi-synthetic fatty acid esters, propylene glycol stearate, guanosine, fractionated palm oil, fractionated coconut oil, Brazil Palm wax, glyceryl tripalmitate, glyceryl tristearate, hydrogenated vegetable oil, cocambum and/or semi-synthetic glycerides.

The protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to any one of claims 1 to 3, wherein the bacteriostatic agent is one or more of the following components. : sorbic acid, phenol, cresol, chlorocresol, propionic acid, benzoic acid, dehydroacetic acid, methylparaben, ethylparaben and/or propylparaben.

11. A protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to any one of claims 1 to 3, wherein the protein protectant is one or more of the following components. Species: trehalose, sucrose, mannitol, histidine, glycine, methionine, sodium tartrate, sodium succinate, cyclodextrin, disodium edetate, human serum albumin, gelatin, sodium chloride and/or Glucan.

12. A protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to any one of claims 1 to 3, wherein the pH adjuster is one or more of the following components. Species: Hydrochloric acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, citric acid, acetic acid, sodium acetate, maleic acid, sodium maleate, sodium citrate, boric acid, sodium borate, citric acid and/or citric acid sodium.

The protein drug rectal suppository which achieves systemic therapeutic effect by local administration of the rectum according to any one of claims 1 to 3, wherein the antioxidant is one or more of the following components. : Nibble methyl ester, Nibel gold ethyl ester, Nipa gold propyl ester, sorbic acid, benzoic acid and salts, tartaric acid and salts, chlorobutanol, thimerosal, ascorbic acid, tocopherol and/or thio Sodium sulfate.

14. A method for preparing a protein drug rectal suppository which achieves systemic therapeutic effect by local administration of rectal administration according to any one of claims 1 to 13, wherein the suppository base is a water-soluble suppository base comprising the following steps: The bacteriostatic agent, the absorption enhancer, the antioxidant and the pH adjuster are mixed and heated according to a certain proportion of the suppository substrate, and cooled to 35~60 ° C plus Into protein drugs and protein protectants, filling, cooling, molding, demoulding, packaging, that is.

The method for preparing a protein drug rectal suppository which achieves systemic therapeutic effect by local administration of rectal administration according to any one of claims 1 to 13, wherein when the suppository base is a fat-soluble suppository base, the following steps are included: First, the fat-soluble matrix is melted at 65-75 ° C, and the absorbent, antioxidant bacteriostatic agent and pH adjuster are separately added to the molten matrix. After mixing uniformly, the temperature is lowered to 35-6 (TC, added Protein drugs and protein protectants, filling, cooling, molding, demoulding, packaging, that is.

16. A protein drug rectal suppository which achieves systemic therapeutic effect by local administration of rectal administration as described in claim 4, in the preparation of a child suffering from endogenous growth hormone deficiency caused by slow growth, severe burns, and clear Growth hormone deficiency caused by hypothalamic-pituitary disease, significant lack of growth hormone confirmed by two different growth hormone tests, negative nitrogen balance after surgery, and anti-aging drugs.

17. A protein-based rectal suppository for the treatment of chronic hepatitis B, chronic hepatitis C, condyloma acuminata, hairy cell leukemia and chronic granulocytes according to claim 5, wherein a protein drug for rectal suppository is achieved by local administration of rectal administration. Application in leukemia drugs.