阅读说明：本技术 一种稳定的重组人干扰素软膏及其生产方法 (Stable recombinant human interferon ointment and production method thereof ) 是由 刘景会 刘琳琳 王莹 王宇 刘玉林 黄曼 郑全莉 杨红育 富瑞丽 王江林 刘晨 于 2019-10-11 设计创作，主要内容包括：本发明涉及医药制剂领域,具体涉及一种稳定的重组人干扰素软膏及其生产方法,所述软膏以重组人干扰素α2a原液为主要活性成分,与软膏基质混合制备能够获得稳定的重组人干扰素软膏,其能够在低温条件下长期保存,质量好、稳定性佳；本发明的重组人干扰素软膏制备工艺包括水相配制、油相配制、原液稀释、以及基质乳化四个步骤,操作简便,便于实际生产应用。(The invention relates to the field of pharmaceutical preparations, in particular to a stable recombinant human interferon ointment and a production method thereof, wherein the ointment takes a recombinant human interferon alpha 2a stock solution as a main active ingredient, and is prepared by mixing with an ointment matrix to obtain the stable recombinant human interferon ointment which can be stored for a long time under a low-temperature condition, and has good quality and good stability; the preparation process of the recombinant human interferon ointment comprises four steps of water phase preparation, oil phase preparation, stock solution dilution and matrix emulsification, is simple and convenient to operate and is convenient to practical production and application.)

1. A recombinant human interferon alpha 2a ointment is characterized in that: the ointment is prepared by mixing a recombinant human interferon alpha 2a stock solution serving as a main active ingredient with a cream matrix, wherein the cream matrix consists of an excipient, an emulsifier, a stabilizer, a preservative and water, and the excipient consists of glycerol, glyceryl monostearate, glyceryl distearate and white vaseline; the emulsifier is polysorbate 80; the stabilizer is dextran 40; the preservative is ethyl p-hydroxybenzoate.

2. The recombinant human interferon alpha 2a ointment of claim 1, which is characterized in that: preferably, the cream matrix comprises the following components in parts by weight: 20:1:0.15:46.5:10:15: 3.

3. the recombinant human interferon alpha 2a ointment of any one of claims 1 to 2, wherein: the content of interferon alpha 2a in the ointment is 1 x 10 in terms of active mass ratio⁴-5×10⁸IU/g ointment, preferably, the content is 1 × 10⁴-1×10⁵IU per gram of ointment.

4. The recombinant human interferon alpha 2a ointment of any one of claims 1 to 2, wherein: it can be used for treating viral infectious skin diseases.

5. The recombinant human interferon alpha 2a ointment of claim 3, which is characterized in that: the virus infectious skin diseases are facial herpes and genital herpes caused by infection of herpes simplex virus I, II type, condyloma acuminatum caused by human papilloma virus and herpes zoster.

6. The recombinant human interferon alpha 2a ointment of any one of claims 1 to 4, wherein: the ointment can be stored for at least 18 months at 2-8 ℃ in the dark.

7. A method for producing a recombinant human interferon alpha 2a ointment according to any one of claims 1 to 5, which comprises: the production method comprises the following steps:

1) Adding glycerol, dextran 40, ethyl p-hydroxybenzoate and injection water into a stainless steel barrel according to the total weight of 20%, 1%, 0.15% and 46.5%, respectively, fully dissolving and mixing uniformly, and sterilizing at 111-131 deg.C for 60-70 min;

2) Determining IU value of the recombinant human interferon alpha 2a in the recombinant human interferon alpha 2a stock solution according to a standard method, and diluting the recombinant human interferon alpha 2a stock solution by 0.10-0.11M phosphate buffer solution with pH of 7.0-7.5 after sterilization according to the IU value of the stock solution and the IU value of the human interferon alpha 2a in the ointment preparation, wherein the amount of the sterilized recombinant human interferon alpha 2a stock solution is 4.35% of the total weight;

3) Weighing white vaseline, glyceryl monostearate, glyceryl distearate and polysorbate-80 respectively according to the amount of 10%, 15% and 3% of the total weight for later use;

4) Setting the temperature of a vacuum homogenizing emulsifying machine to be 81-83 ℃, stirring at the speed of 20-25 rpm, adding white vaseline into the emulsifying machine while stirring, adding glycerin monostearate and glycerin distearate after melting, adding polysorbate-80 after fully melting, uniformly mixing the sterilized water phase, slowly pouring the uniformly mixed water phase into the emulsifying machine, continuously stirring and uniformly mixing, opening a cooling circulating water switch, reducing the temperature to 52-56 ℃, adding the recombinant human interferon alpha 2a diluent obtained in the step 2), and continuously stirring until complete emulsification and sealing for later use;

5) connecting a silica gel pipeline with a discharge port of the vacuum homogenizing emulsifying machine and a feeding port of a feeding tank of a filling machine, connecting compressed air to a top connecting port of the vacuum homogenizing emulsifying machine, opening the compressed air and the discharge port, and transferring the ointment matrix into the feeding tank by using the compressed air;

6) Setting the temperature of a material feeding tank of an ointment filling and end sealing machine to be 52-56 ℃, subpackaging at a speed of 60-70 counts/min, and filling the medicinal polyethylene composite ointment pipe after the air of a filling head is exhausted;

7) Setting the temperature of a heat-sealing tail of an ointment filling and tail-sealing machine to be 350-360 ℃, heating and sealing the tail, marking a batch number, cutting into a product to be packaged, and finally packaging into a finished product after the product is qualified.

Technical Field

The invention relates to the field of pharmaceutical preparations, in particular to a stable recombinant human interferon ointment and a production method thereof

Background

The recombinant human interferon alpha 2a is a kind of multifunctional and high activity inducing protein produced by leucocyte and lymphocyte and is unstable in light and moisture. The recombinant human interferon alpha 2a has broad-spectrum antiviral action and immunoregulation function on organisms, and the antiviral mechanism is mainly realized by combining interferon with a target cell surface interferon receptor, inducing multiple antiviral proteins such as 2-5(A) synthetase, protein kinase PKR, MX protein and the like in a target cell, preventing the synthesis of viral protein and inhibiting the replication and transcription of viral nucleic acid.

The development of the dosage form of the recombinant human interferon alpha 2a is not ideal all the time, and the application of the recombinant human interferon alpha 2a is influenced. The recombinant human interferon alpha 2a clinically used at present is an injection, a tablet, a gel, a cream, a suppository and the like. The higher the interferon concentration, the better the effect of treating the virus. However, some dosage forms of recombinant human interferon alpha 2a have great influence on the treatment effect due to various side effect reactions caused by systemic administration or local administration with overhigh concentration. The commonly used recombinant human interferon alpha 2a injection has the problems of inconvenient use and slight pain, can not directly act on the focus, has limited concentration reaching the application part and has certain adverse reaction. After the recombinant human interferon alpha 2a tablet enters the body, the recombinant human interferon alpha 2a is difficult to dissolve out and play a role. Although the recombinant human interferon alpha 2a suppository has good curative effect on treating female chronic cervicitis, cervical erosion and vaginitis caused by virus infection, the suppository also has limitations, and the clinical curative effect of the suppository is mainly related to the biological activity of the recombinant human interferon alpha 2 a.

The existing human interferon gel and cream have the defects that liposome needs to be prepared or human serum albumin needs to be used as a raw material, so that the preparation process is complex, the risk of virus infection exists, the actual production operation is not facilitated, and the production cost is high due to the human serum albumin, more raw materials and operation steps.

Therefore, aiming at the defects of the prior art, the invention further researches and develops the recombinant human interferon alpha 2a ointment and the preparation method thereof.

Disclosure of Invention

In order to solve the defects of the prior art, the invention provides a recombinant human interferon alpha 2a ointment which is prepared by uniformly mixing a recombinant human interferon alpha 2a stock solution obtained by fermentation and purification with a matrix prepared from glycerol, dextran 40, ethyl p-hydroxybenzoate, white vaseline, glycerin monostearate, polysorbate-80 and water for injection according to a proportion.

Specifically, the invention provides a recombinant human interferon alpha 2a ointment, which is prepared by mixing a recombinant human interferon alpha 2a stock solution serving as a main active ingredient with a cream matrix, wherein the cream matrix consists of an excipient, an emulsifier, a stabilizer, a preservative and water, and the excipient consists of glycerol, glycerol monostearate, glycerol distearate and white vaseline; the emulsifier is polysorbate 80; the stabilizer is dextran 40; the preservative is ethyl p-hydroxybenzoate.

Further, the preferable cream matrix comprises glycerin, dextran 40, ethyl p-hydroxybenzoate, water for injection, white vaseline, glyceryl monostearate, glyceryl distearate and polysorbate-80 in parts by weight: 20:1:0.15:46.5:10:15:3.

Furthermore, the content of interferon alpha 2a in the ointment is 1 x 10 in terms of active mass ratio⁴-5×10⁸IU/g ointment, preferably, the content is 1 × 10⁴-1×10⁵IU per gram of ointment.

Further, the ointment can be used for treating viral infectious skin diseases.

Preferably, the viral infectious skin diseases are facial herpes and genital herpes caused by herpes simplex virus type i and type ii infections, condyloma acuminatum and herpes zoster caused by human papilloma virus.

Furthermore, the ointment can be stably stored for at least 18 months under the condition of being kept at 2-8 ℃ in the dark.

The invention also provides a production method of the recombinant human interferon alpha 2a ointment, which comprises the following steps:

1) Adding glycerol, dextran 40, ethyl p-hydroxybenzoate and injection water into a stainless steel barrel according to the total weight of 20%, 1%, 0.15% and 46.5%, respectively, fully dissolving and mixing uniformly, and sterilizing at 111-131 deg.C for 60-70 min;

2) Determining IU value of the recombinant human interferon alpha 2a in the recombinant human interferon alpha 2a stock solution according to a standard method, and diluting the recombinant human interferon alpha 2a stock solution by 0.10-0.11M phosphate buffer solution with pH of 7.0-7.5 after sterilization according to the IU value of the stock solution and the IU value of the human interferon alpha 2a in the ointment preparation, wherein the amount of the sterilized recombinant human interferon alpha 2a stock solution is 4.35% of the total weight;

3) Weighing white vaseline, glyceryl monostearate, glyceryl distearate and polysorbate-80 respectively according to the amount of 10%, 15% and 3% of the total weight for later use;

4) Setting the temperature of a vacuum homogenizing emulsifying machine to be 81-83 ℃, stirring at the speed of 20-25 rpm, adding white vaseline into the emulsifying machine while stirring, adding glycerin monostearate and glycerin distearate after melting, adding polysorbate-80 after fully melting, uniformly mixing the sterilized water phase, slowly pouring the uniformly mixed water phase into the emulsifying machine, continuously stirring and uniformly mixing, opening a cooling circulating water switch, reducing the temperature to 52-56 ℃, adding the recombinant human interferon alpha 2a diluent obtained in the step 2), and continuously stirring until complete emulsification and sealing for later use;

5) connecting a silica gel pipeline with a discharge port of the vacuum homogenizing emulsifying machine and a feeding port of a feeding tank of a filling machine, connecting compressed air to a top connecting port of the vacuum homogenizing emulsifying machine, opening the compressed air and the discharge port, and transferring the ointment matrix into the feeding tank by using the compressed air;

6) Setting the temperature of a material feeding tank of an ointment filling and end sealing machine to be 52-56 ℃, subpackaging at a speed of 60-70 counts/min, and filling the medicinal polyethylene composite ointment pipe after the air of a filling head is exhausted;

7) Setting the temperature of a heat-sealing tail of an ointment filling and tail-sealing machine to be 350-360 ℃, heating and sealing the tail, marking a batch number, cutting into a product to be packaged, and finally packaging into a finished product after the product is qualified.

the specific production process of the invention comprises the following steps: preparing water phase → sterilizing → diluting stock solution → preparing oil phase → emulsifying matrix → transferring matrix → filling → sealing tail → cutting to prepare product to be packaged → testing qualified → packaging to obtain finished product.

Advantageous effects

The matrix formula of the invention is particularly suitable for the characteristics of recombinant human interferon alpha 2a, and is specifically embodied as follows:

dextran 40 is a plasma substitute solution, can reduce blood viscosity and improve microcirculation except the function of expanding blood volume, has the properties of human serum albumin in physicochemical properties, and has the functions of protecting and stabilizing interferon structure and activity.

The glyceryl monostearate and the glyceryl distearate are nonionic emulsifiers, are water-in-oil type, have both hydrophilic groups and lipophilic groups, so that the interferon stock solution can be better dissolved in a matrix, the ointment is more exquisite and smooth, and compared with an oily emulsifier, the skin can be absorbed and cleaned more easily.

Polysorbate-80 is used as nonionic surfactant and transdermal absorption enhancer to promote skin to absorb interferon so as to further increase drug effect.

The ethyl p-hydroxybenzoate not only serves as a preservative and a bacteriostatic agent, but also can increase the water solubility of the matrix when being used together with polysorbate-80, so that the interferon can exert the drug effect more conveniently.

White vaseline has water resistance, can keep skin moist when being smeared on the skin, enables skin tissues of wound parts to keep the optimal state, accelerates the self-repairing capacity of the skin, can better protect the skin, relieves the pain of patients, and plays a role in adjuvant therapy. In addition, white petrolatum can block bacteria from the air from contacting the skin, thereby reducing the likelihood of infection.

glycerol is used as a stabilizer and excipient and has strong water absorption. The physical action of glycerol is as a dehydrating agent, and since bacteria reproduce slowly in a local dehydration environment, glycerol can play a role in stabilizing interferon. And besides moistening the skin, the glycerin also has the effect of effectively protecting damaged skin mucous membranes.

The production process is simple and easy to implement, low in manufacturing cost and suitable for large-scale industrial production, and simultaneously ensures that the product has better quality uniformity and stability.

The invention is used as a local external preparation and directly acts on the local part of a lesion, so that the interferon with low concentration keeps high active ingredients in the damaged skin mucosa, the local medicine concentration is higher, the treatment effect is very obvious, the common cold-like symptoms and other side reaction symptoms after the interferon is taken by the whole body of the product can not occur, and the product has no toxic or side effect.

The invention is a local external preparation with convenient use, safety, no pain, economy and definite curative effect, which is mainly used for treating face herpes and genital herpes caused by herpes simplex virus I and II infection and condyloma acuminatum and herpes zoster caused by human papilloma virus by transdermal administration.

Drawings

FIG. 1: the pH of the recombinant human interferon alpha 2a ointment accelerates the stability test result (23-27 ℃).

FIG. 2: the biological activity of the recombinant human interferon alpha 2a ointment accelerates the stability test result (23-27 ℃).

FIG. 3: the long-term stability test result of the pH of the recombinant human interferon alpha 2a ointment (2-8 ℃).

FIG. 4: the long-term stability investigation result of the biological activity of the recombinant human interferon alpha 2a ointment (2-8 ℃).

Detailed Description

The present invention is described in more detail below to facilitate an understanding of the present invention.

It should be understood that the terms or words used in the specification and claims should not be construed as having meanings defined in dictionaries, but should be interpreted as having meanings that are consistent with their meanings in the context of the present invention on the basis of the following principles: the concept of terms may be defined appropriately by the inventors for the best explanation of the invention.