CN114042036A

CN114042036A - Ganciclovir external preparation and application thereof

Info

Publication number: CN114042036A
Application number: CN202111587952.4A
Authority: CN
Inventors: 刘实; 杨芬芬; 崔琳; 邓姿萍
Original assignee: Beijing Dingshi Pharmaceutical Technology Co ltd
Current assignee: Beijing Dingshi Pharmaceutical Technology Co ltd
Priority date: 2021-12-23
Filing date: 2021-12-23
Publication date: 2022-02-15

Abstract

The invention relates to a ganciclovir external preparation and application thereof. The external preparation is prepared from the following raw materials: ganciclovir, a high molecular matrix and a pH regulator. The ganciclovir external preparation has better antiviral effect than acyclovir gel.

Description

Ganciclovir external preparation and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a ganciclovir external preparation and application thereof.

Background

Herpes simplex virus type II (HSV-2), a typical member of the alphaherpesviridae subfamily, can infect the epithelium causing genital herpes, latent in the peripheral nervous system and spread to the central nervous system, resulting in a lifelong latent infection. More than about 4 million people worldwide are infected with this virus, and HSV-2 infection is currently incurable, creating a significant medical and public health problem.

In the clinical treatment of herpes virus, mainly nucleoside analogues (acyclovir or valacyclovir) are taken as main drugs, the nucleoside analogues stop ongoing DNA chain elongation by inhibiting viral DNA polymerase, can effectively resist HSV-2 infection and reduce the severity and duration of symptoms, but cannot completely cure the herpes virus, and latent viruses still can be reactivated when the immune state is disordered to cause irregular relapse of the symptoms. The existing drug acyclovir for treating herpes virus in the market has dosage forms of gel, cream and the like, and also has the problems.

Therefore, there remains a clinical need for new effective anti-herpes drugs.

Disclosure of Invention

For the reasons, the applicant obtains a new ganciclovir external preparation through multiple creative tests, wherein the external preparation is prepared from the following raw materials: ganciclovir, a high molecular matrix and a pH regulator. Experimental research shows that the ganciclovir external preparation has better antiviral effect than acyclovir gel.

The invention is realized by the following technical scheme.

In one aspect, the invention provides a ganciclovir external preparation, which is prepared from the following raw materials: ganciclovir, a high molecular matrix, a pH regulator and the balance of water.

Preferably, the external preparation is prepared from the following raw materials in percentage by weight: 0.15-3% of ganciclovir, 0.3-0.4% of polymer matrix, 0.1-1% of pH regulator and the balance of water;

more preferably, the content of ganciclovir in the external preparation is 0.15-1%, preferably 0.15-0.5% by weight;

more preferably, the content of the polymer matrix in the external preparation is 0.35-0.4% by weight, preferably 0.4% by weight. Preferably, the polymer matrix is selected from one or more of carbomer, xanthan gum, cetyl alcohol, stearyl alcohol, glyceryl stearate, white vaseline, liquid paraffin, lanolin, hydrogenated castor oil, dimethicone and polyethylene glycol;

preferably, the polymeric matrix is carbomer.

Preferably, the pH adjusting agent is selected from one or more of sodium hydroxide, potassium hydroxide, triethanolamine, ethylenediamine, sodium bicarbonate, and amino acids;

preferably, the pH adjuster is sodium hydroxide or triethanolamine.

Preferably, the external preparation further comprises one or more of the following raw materials: humectant, antiseptic, and skin penetration enhancer.

Preferably, the humectant is selected from glycerin, propylene glycol;

preferably, the preservative is selected from ethylparaben, benzalkonium chloride;

preferably, the transdermal absorption enhancer is selected from laurocapram.

Preferably, the pH value of the external preparation is 6.0-7.5;

preferably, the external preparation is a gel.

In another aspect, the present invention also provides the use of the aforementioned external preparation for the preparation of an antiviral drug.

Preferably, the antiviral drug is an anti-herpes virus drug.

Preferably, the antiviral drug is an anti-herpes simplex virus type II drug.

At present, no ganciclovir external preparation exists in the market, the ganciclovir external preparation fills the gap of the market, and the antiviral effect of the ganciclovir external preparation is superior to that of the existing acyclovir external preparation in the market.

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

fig. 1 is a graph of herpes score data for the external preparation of the present invention and acyclovir gel.

Fig. 2 is a graph showing skin score data of the external preparation of the present invention and acyclovir gel.

Fig. 3 is a graph of clinical symptom score data of the external preparation of the present invention and acyclovir gel.

Detailed Description

Specific embodiments of the present invention will be described in more detail below with reference to embodiments and the accompanying drawings. While specific embodiments of the invention are shown in the drawings, it should be understood that the invention may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

It should be noted that certain terms are used throughout the description and claims to refer to particular combinations. It will be understood by those skilled in the art that various terms may be used to refer to a single composition. The present specification and claims do not differentiate between components by noun differences, but rather combine functional differences as criteria for differentiation. In the following description and in the claims, the terms "include" and "comprise" are used in an open-ended fashion, and thus should be interpreted to mean "include, but not limited to. The description which follows is a preferred embodiment of the invention, but is made for the purpose of illustrating the general principles of the invention and not for the purpose of limiting the scope of the invention. The scope of the present invention is defined by the appended claims.

For the purpose of facilitating an understanding of the embodiments of the present invention, the following description will be made in terms of several specific embodiments with reference to the accompanying drawings, and the drawings are not intended to limit the embodiments of the present invention.

The research on the component types, the dosage and the pH value in the external preparation of the invention

EXAMPLE 1.1 selection of the type of adjuvant

In the development process, the inventor performs a compatibility test of raw and auxiliary materials, wherein table 1 is a raw and auxiliary material ratio, and table 2 is a stability test.

TABLE 1 raw and auxiliary materials ratio

The preparation method comprises the following steps of respectively preparing raw materials and auxiliary materials according to the raw material and auxiliary material proportion in the table 1, uniformly mixing the raw materials and the auxiliary materials, mixing the solid oil phase matrix with API (namely ganciclovir) after melting, directly mixing the liquid matrix, and mixing the water phase solid auxiliary material preparation with the API. And (3) respectively placing the mixed materials under the conditions of illumination and high temperature of 40 ℃, and sampling and detecting after 10 days. The results are shown in Table 2.

TABLE 2 stability test results

The result shows that under the conditions of illumination and high temperature, the properties of the sample are changed after the sample is placed for 10 days, under the condition of high temperature, the related substances of cetearyl alcohol, lanoline and the API mixture are increased, and the properties and related substances of other raw and auxiliary material mixtures are not obviously changed, so that the incompatibility of hypromellose, cetearyl alcohol and ganciclovir is shown, and other auxiliary materials can be selected for research of the product.

EXAMPLE 1.2 selection of adjuvant dosage-screening of the amount of polymeric matrix (carbomer)

This example, using carbomer as an example, investigated the effect of dosage on gel formulations, screened by the formula of table 3.

TABLE 3

The preparation method comprises the following steps: (1) adding carbomer in a formula amount into purified water accounting for 40% of the total weight, and stirring to fully swell to form uniform dispersion liquid;

(2) dissolving sodium hydroxide, ganciclovir, benzalkonium chloride and glycerol by using purified water accounting for 50% of the total weight, adding the filtrate into the dispersion liquid obtained in the step (1) while stirring;

(3) regulating pH to about 7 with triethanolamine, adding purified water to a sufficient amount (100%), and stirring to form uniform gel.

Observing or measuring the properties, pH value and viscosity of the prepared product. The results are shown in Table 4.

TABLE 4

As can be seen from table 4, the amount of carbomer added is preferably 0.35% to 0.4%, more preferably 0.4%, since carbomer is added in an amount of 0.5%, has an excessively high gel viscosity, is in the form of water after being coated at a concentration of 0.3%, and has a weak adhesive force.

EXAMPLE 1.3 determination of the pH Range of the gels

The external preparation belongs to a semisolid preparation, and the pH value is a key quality attribute, so the pH value range of the external preparation is screened to determine a proper pH value range capable of stabilizing the preparation, the pH value of human skin is about 5.5-7.0, and ganciclovir is unstable under an acidic condition, so the range of the external preparation is 5.5-7.5. The formulations used are shown in table 5.

TABLE 5

(3) regulating pH value to preset pH value with triethanolamine, adding purified water to reach sufficient amount (100%), and stirring to form homogeneous gel;

(4) and filling the prepared ganciclovir gel into a medicinal aluminum ointment tube to obtain the ganciclovir gel.

Placing the sample at the conditions of high temperature of 60 ℃, illumination of 5000 +/-500 Lux and low temperature of 2-8 ℃, taking out the sample after 10 days, and detecting, wherein the results are shown in the following table 6:

TABLE 6

As can be seen from the above table, after the samples under various pH conditions are placed under the conditions of high temperature of 60 ℃, low temperature of 2-8 ℃ and illumination for 10 days, the properties, pH values, contents and related substances of the gel preparation with pH of 6.0-7.5 are not obviously changed, the related substances of the samples are slightly increased under the condition of pH5.5, and according to the test results, the pH value range of the product is 6.0-7.5.

Example 1.4 selection of Ganciclovir dosage

The ganciclovir molecular structure is similar to that of acyclovir, the ganciclovir external preparation is an antiviral drug, the concentration of the existing ganciclovir external preparation on the market is 3%, the ganciclovir external preparation on the market is an eye preparation, the general concentration is 0.15%, the ganciclovir external preparation is a ganciclovir external preparation, and the ganciclovir external preparation is compared with an acyclovir preparation for research, so that the ganciclovir preparation specification of the ganciclovir external preparation is selected from a range of 0.15% -3% for research. Ganciclovir is first dissolved completely in water solution and then added to gel matrix. The recipes used are shown in table 7.

TABLE 7

(3) adjusting pH with triethanolamine or hydrochloric acid water solution, adding purified water to reach sufficient amount (100%), and stirring to form uniform gel.

After 10 days of storage under high temperature and light conditions, the gel formulations of ganciclovir at 0.15% -3% concentration had the following results in table 8:

TABLE 8

According to the test result, the pH value of the 4.0% ganciclovir needs to be adjusted back because the pH value is too high and is not gelatinous after being mixed with the carbomer dispersion liquid due to excessive addition of sodium hydroxide during preparation, and the process is relatively more complex. After the ganciclovir gel preparation with the concentration of 0.15-3% is placed for 10 days under the conditions of high temperature and illumination, the properties, the pH value and related substances do not change obviously, the influence factor result is good, but in order to dissolve ganciclovir in the preparation process, the addition of sodium hydroxide in the ganciclovir concentration formulas with the concentration of 2-3% causes the pH value of the gel to be higher, and according to the pharmacodynamic result, the efficacy of ganciclovir is obviously superior to that of acyclovir preparations, the clinical requirement can be completely met, and high-concentration preparations do not need to be prepared, so the ganciclovir concentration of the invention is preferably 0.15-1.0%, more preferably 0.15-0.5%.

Preparation examples

Example 2.1:

TABLE 9 raw and auxiliary Material compositions of example 2.1

Composition (I)	Dosage (w/w%)
		Ganciclovir	0.15
Carbomer	0.4
		Benzalkonium chloride	0.0075
Glycerol	3.0
		Sodium hydroxide	0.08
Triethanolamine	Proper amount of
		Purified water (added to total 100)	Proper amount of
In total	100

(3) regulating the pH value to 6.0-7.5 by using triethanolamine, adding purified water to a sufficient amount (100%), and stirring to form uniform gel;

Example 2.2:

TABLE 10 raw and auxiliary materials composition of example 2.2

Example 2.3:

table 11 raw and auxiliary material composition of example 2.3:

composition (I)	Dosage (w/w%)
		Ganciclovir	1.0
Carbomer	0.4
		Benzalkonium chloride	0.0075
Glycerol	3.0
		Sodium hydroxide	0.16
Triethanolamine	Proper amount of
		Purified water (added to total 100)	Proper amount of
In total	100

Stability test

Example 3.1 stability Studies

The samples prepared in example 2.1-example 2.4 were observed at 30 ℃. + -. 2 ℃ and RH 65. + -. 5% for 6 months, and sampled and tested at 1, 2, 3 and 6 months, respectively, and the results are shown in Table 12 below:

TABLE 12 results of stability studies

The results show that: the accelerated test is carried out for 6 months, the properties, the content, the pH value and related substances of the samples of 3 groups of examples have no obvious change, the granularity of the raw materials in the samples accords with the specification of pharmacopoeia, and the stability of the samples is good.

Animal testing

Example 4.1 Observation of therapeutic efficacy of the external preparation ganciclovir gel of the invention in treating guinea pig type II herpes simplex virus (HSV 2)

1. Material

(1) Virus: herpes simplex virus type II (HSV 2), is an amplification of Mingkanded pharmaceutical science and technology, Inc., Nantong medicine.

(2) Guinea pigs: female guinea pigs weighing 250-300 g were purchased from Beijing Wittiulihua laboratory animal technologies, Inc.

(3) Medicine preparation: comparison: acyclovir gel (containing main drug acyclovir 1.0%, available from Shandongming pharmaceutical group, Inc., 2007081, 12 months 2020), self-made: ganciclovir gel (containing ganciclovir 0.15%, 0.5%, i.e. example 2.1 and example 2.2), vehicle.

2. Method of producing a composite material

(1) All guinea pigs were weighed before inoculation and then subjected to a random group, with no significant difference in mean body weight between groups.

(2) Guinea pigs were vaccinated with the virus via dermabrasion on day 0 at a dose of 4.08E +05 p.f.u./guinea pig at a volume of 0.02mL on one side and 0.04mL on both sides.

(3) Animal grouping: guinea pigs were divided into 4 groups, group 1: vehicle group, group 2: acyclovir gel (positive control) group, group 3: 0.15% ganciclovir gel group, group 4: a 0.5% ganciclovir gel group, 5 guinea pigs per group.

(4) The method comprises the following steps: continuously treating the raw materials with acyclovir gel, ganciclovir gel with different concentrations and blank solvent for 7 days from day 1 to day 7 respectively, wherein the administration interval is 6/6/12 hours every day, the administration mode is smearing, the administration volume is 0.5g, the administration is carried out for 21 times, and the first administration time is 24 hours after virus inoculation. Weighing, dosing and health monitoring of experimental animals on days 0 to 7, weighing and health monitoring of experimental animals on days 8 to 14, and end point of experiment: the surviving animals were euthanized.

(5) The detection indexes of the curative effect are as follows:

herpes scoring: no herpes or herpes scab shedding: 0 minute; 1-5 herpes: 1 minute; 5-15 herpes: 2 min; more than 15 are provided: 3 min;

skin disorder scoring: slight red and swollen is 1 point, red and swollen is 2 points, red and swollen of thigh and vulva is 3 points, and severe red and swollen is 4 points.

(6) Statistical analysis: analysis of body weight changes and herpes clinical morbidity scores was performed using Two Way ANOVA.

(7) Inclusion criteria for experimental results:

vehicle group (group 1): the incidence rate of the diseases (with herpes) is more than or equal to 60 percent.

If the experimental result meets the above conditions, the experimental data is considered to be credible, and the experimental result is accepted.

3. As a result:

(1) weight change:

vehicle group 1: the average weight of the guinea pigs after infection is relatively stable, obvious weight loss occurs until day 7, the weight begins to rise again until day 9, and the weight of the guinea pigs rises by 11% at the end of the experiment;

group 2 acyclovir gels: the average weight of the guinea pigs continuously increased after infection, slightly decreased on day 7, and started to rise again after day 8, and the weight of the guinea pigs increased by 20% at the end of the experiment;

group 3, 0.15% ganciclovir gel: the weight of the guinea pig shows a rising trend after infection, and the weight of the guinea pig rises by 30% at the end point of the experiment;

group 4, 0.5% ganciclovir gel: guinea pigs showed a tendency to gain weight after infection, with the end point of the experiment showing a 30% increase in weight.

(2) The incidence of diseases is as follows:

vehicle group 1: the animals developed herpes on day 4 with 100% incidence on day 5 (6/6).

Group 2 acyclovir gel group: the animals developed herpes on day 5 with a 50% incidence by the end of the experiment (3/6).

Group 3, 0.15% ganciclovir gel group: the animals developed herpes on day 5 with a prevalence of 16.66% by the end of the experiment (1/6).

Group 4, 0.5% ganciclovir gel group: the incidence of the animals was 0% by the end of the experiment (0/6).

(3) Herpes score (number of herpes)

Vehicle group 1: the herpes score increased from day 4, reached a maximum of 3 on

day

7 and 8, declined from day 9 to day 13 with a herpes disappearance score of 0.

Group 2 acyclovir gel group: the score started to increase from day 5, the mean values on

days

7 and 8 reached the highest value of 0.8, the score started to decrease from day 9 to day 13, and the herpes vanishing score was 0; after day 4, the symptoms were significantly better than the vehicle group.

Group 3, 0.15% ganciclovir gel group: the herpes score started to rise on day 5, with a 6 th day balance of 0.33 maximum and a herpes disappearance score of 0 by day 10; after day 4, the symptoms were significantly better than the vehicle group.

Group 4, 0.5% ganciclovir gel group: from the start of day 0 to the end of the experiment, no herpes was observed and scored as 0. After day 4, the symptoms were significantly better than the vehicle group. The results are shown in FIG. 1.

(4) Skin scoring

Vehicle group 1: the skin score increased from day 5 to the 7 th day when the mean reached the highest value of more than 3 points, after which the mean fluctuated over 2;

group 2 acyclovir gel group: the skin score increased from day 5 to a peak of 1.2 on the 7 th mean and then decreased to a redness disappearance score of 0 by day 13; after day 6, the symptoms were significantly better than the vehicle group.

Group 3, 0.15% ganciclovir gel group: the skin score increased from day 5 to a maximum of 1.2 on average at day 6, followed by a rapid decline with a red swelling disappearance score of 0 by day 13; after day 7, the symptoms were significantly better than the vehicle group.

Group 4, 0.5% ganciclovir gel group: the skin score increased from day 6, with the mean values at

day

6 and 7 reaching a maximum of 0.7 points, followed by a rapid decline until day 8 with a redness-swelling disappearance score of 0; after day 6, the symptoms were significantly better than the vehicle group. The results are shown in FIG. 2. (5) Clinical symptom score (skin red swelling and herpes combined score)

Vehicle group 1: clinical symptom scores increased from day 4, reached a maximum of 6 points on the 7 th mean, then dropped to 1.3 points on the 12 th mean. The number of the clinical symptoms is 1.5 when the number of the clinical symptoms is increased by 13 days and 14 days after the experiment is finished;

group 2 acyclovir gel group: clinical symptom score increased from day 5 to mean 2 points at day 7, followed by a decline to day 13 score of 0; after day 5, the symptoms were significantly better than the vehicle group.

Group 3, 0.15% ganciclovir gel group: clinical symptom score increased from day 5 to a maximum of 1.5 on day 6, followed by a rapid decline to day 13 score of 0; after day 5, the symptoms were significantly better than the vehicle group.

Group 4, 0.5% ganciclovir gel group: clinical symptom scores increased from day 6, mean values reached a maximum of 0.7 on

days

6 and 7, 0 on day 8, no other symptoms by day 14 after the end of the experiment, and 0. After day 5, the symptoms were significantly better than the vehicle group. The results are shown in FIG. 3.

4. Conclusion

The inventor utilizes a guinea pig pathogenic model of HSV infection to observe the treatment effect of ganciclovir gel on guinea pig herpes virus, and finds that the medicine has an obvious HSV resistant effect, which is shown in that ganciclovir can reduce the degree of HSV lesion and accelerate the healing of the lesion. On the 4 th day of treatment, the treatment effect is shown, on the 5 th day of treatment, the treatment effect is better in the 0.15 percent and 0.5 percent ganciclovir groups and the acyclovir group, and the antiviral effect of the ganciclovir gel is obviously better than that of the acyclovir gel.

The experimental result shows that the ganciclovir gel has the therapeutic effect on the herpes virus and is a new dosage form for resisting the herpes virus with hopeful transition to clinic.

It should be understood that the above examples are only for clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. It is intended that the present invention be understood and implemented by those skilled in the art, and not limited thereto. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. The ganciclovir external preparation is prepared from the following raw materials: ganciclovir, a high molecular matrix, a pH regulator and the balance of water.

2. The external preparation according to claim 1, wherein the external preparation is prepared from the following raw materials in percentage by weight: 0.15-3% of ganciclovir, 0.3-0.4% of polymer matrix, 0.1-1% of pH regulator and the balance of water.

3. The external preparation according to claim 1 or 2, wherein the content of ganciclovir in the external preparation is 0.15-1%, preferably 0.15-0.5%;

preferably, the content of the polymer matrix in the external preparation is 0.35-0.4% by weight, and preferably 0.4% by weight.

4. The external preparation according to any one of claims 1 to 3, wherein the polymeric base is selected from one or more of carbomer, xanthan gum, cetyl alcohol, stearyl alcohol, glyceryl stearate, white petrolatum, liquid paraffin, lanolin, hydrogenated castor oil, dimethicone, polyethylene glycol; preferably, the polymeric matrix is carbomer.

5. The external preparation according to any one of claims 1 to 4, wherein the pH-adjusting agent is selected from one or more of sodium hydroxide, potassium hydroxide, triethanolamine, ethylenediamine, sodium bicarbonate, and amino acids; preferably, the pH adjuster is sodium hydroxide or triethanolamine.

6. The external preparation according to any one of claims 1 to 5, wherein the external preparation further comprises one or more of the following raw materials: humectant, antiseptic, skin penetration enhancer;

preferably, the humectant is selected from glycerin and propylene glycol;

preferably, the preservative is selected from ethylparaben and benzalkonium chloride;

preferably, the transdermal absorption enhancer is selected from laurocapram.

7. The external preparation according to any one of claims 1 to 6, wherein the pH of the external preparation is 6.0 to 7.5;

preferably, the external preparation is a gel.

8. Use of the external preparation according to any one of claims 1 to 7 for the preparation of an antiviral medicament.

9. The use of claim 8, wherein the antiviral drug is an anti-herpes virus drug.

10. The use of claim 8, wherein the antiviral drug is an anti-herpes simplex II virus drug.