阅读说明：本技术 包含淫羊藿苷元和羟基积雪草酸的药物组合物的医药用途 (Medical application of pharmaceutical composition containing icaritin and madecassic acid ) 是由 张贵民 李兰芳 高雷 于 2020-11-04 设计创作，主要内容包括：本发明属于医药领域,具体涉及一种药物组合物的医药用途。所述药物组合物由淫羊藿苷元和羟基积雪草酸组成。本发明还公开了所述药物组合物在制备治疗银屑病药物中的应用。本发明药物组合物治疗银屑病的效果不仅显著优于两种药物单独使用时的效果,更优于两者效果的加和；两种药物联合使用可以明显减少各自用量,减少不良反应发生；同时,增加银屑病患者的临床用药选择,有很好的临床应用前景。(The invention belongs to the field of medicines, and particularly relates to a medical application of a pharmaceutical composition. The pharmaceutical composition consists of icaritin and madecassic acid. The invention also discloses application of the pharmaceutical composition in preparing a medicament for treating psoriasis. The effect of the pharmaceutical composition for treating psoriasis is not only obviously superior to the effect of two drugs when used separately, but also superior to the addition of the two effects; the two medicines are combined for use, so that the respective dosage can be obviously reduced, and adverse reactions are reduced; meanwhile, the clinical medication selection of patients with psoriasis is increased, and the medicine has good clinical application prospect.)

1. A pharmaceutical composition containing icariin and madecassic acid is used for preparing medicine for treating psoriasis.

2. Use according to claim 1, characterized in that: the medicine is used for treating psoriasis vulgaris, joint psoriasis, erythrodermic psoriasis and pustular psoriasis.

3. Use according to claim 1, characterized in that: the weight ratio of the icariin to the madecassic acid in the pharmaceutical composition is 1: 0.01-30.

4. Use according to claims 1-3, characterized in that: the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or auxiliary material.

5. Use according to claims 1-3, characterized in that: the madecassic acid in the pharmaceutical composition is madecassic acid, or a salt or a complex thereof.

6. Use according to claims 1-3, characterized in that: the pharmaceutical composition can be prepared into clinically acceptable dosage forms.

7. Use according to claim 6, characterized in that: the clinically acceptable preparation formulation is an oral preparation, an injection preparation or an external preparation.

8. Use according to claim 7, characterized in that the oral formulation is one or more of a tablet, capsule or microemulsion formulation thereof.

9. Use according to claim 7, characterized in that the injectable preparation is one or more of an injection or an injectable microemulsion thereof.

10. The use according to claim 7, wherein the external preparation is one or more of a powder, an ointment, a gel, a cream, a spray or a patch, and the external preparation is administered through the skin.

Technical Field

The invention belongs to the field of medicines, relates to a medical application of a pharmaceutical composition, and particularly relates to an application of a composition containing icariin and madecassic acid in preparation of a medicine for treating psoriasis.

Background

The icariin belongs to flavonol compounds, is slightly present in epimedium medicinal materials, and has the following chemical structural formula:

icariin can be isolated from epimedium medicinal material (grandson, xu Ying, Wen bright light, etc., chemical components of epimedium koreanum, China journal of phytochemistry 1998, 8 (2): 122-125), or icariin can be isolated by enzymolysis (leaf spergualin, Liujia, Loujia, preparation of icariin derivatives and studies on estrogen-like action thereof, proceedings of Zhejiang university, 2005, 34 (2): 131-136).

There is a report that icaritin has the effect of resisting apoptosis of mouse primary culture nerve cells caused by A beta peptide (Zhang Annan, Wang Huanhuan, Wang Zhi Qiang, etc., icaritin has the effect of resisting apoptosis of mouse primary culture nerve cells caused by A beta peptide, Zhejiang university report, 2007, 36 (3): 224-226). Chinese patent CN101836976A discloses that icaritin has the function of resisting tumor angiogenesis. Chinese patent CN101428015A discloses that icaritin has the effect of resisting endotoxemia. Chinese patent CN101284000A discloses that icariin has the function of preventing and treating obesity or fatty liver. Chinese patent CN1869204A discloses the use of icariin in inducing stem cell in vitro directional differentiation. Chinese patent CN1194701C discloses the application of icariin or demethylicariin in the preparation of estrogen receptor modulators.

Madecassic acid is an important pentacyclic triterpenic acid compound in centella asiatica of Umbelliferae, and has the following chemical structural formula:

recent studies have shown that asiatic acid and madecassic acid have certain effects in resisting oxidation and cancer. Chinese patent CN201110040181.7 discloses that madecassic acid has obvious inhibitory effect on mammary gland hyperplasia of experimental mice. Chinese patent CN200910067429.1 discloses the use of asiatic acid and madecassic acid in the preparation of drugs for alpha-glucosidase inhibitors.

The content of aglycone such as madecassic acid in centella asiatica is much lower than that of madecassic acid glycoside, the content of saponin in centella asiatica is generally 1.8-5%, and the content ratio of sapogenin to saponin is generally 1: 2.5. chinese patent CN201610728601.3 discloses a method for preparing madecassic acid by hydrolyzing madecassoside. The method can rapidly break glycosidic bond at normal temperature and normal pressure, has mild hydrolysis process and thorough hydrolysis, and does not generate configuration change of madecassic acid.

Psoriasis is a common chronic, recurrent inflammatory skin disease. The disease has a relatively long course, and cannot be completely cured at present. As an erythrosquamous dermatosis, it often causes impairment of the appearance of the patient and seriously affects the quality of life and even physical and mental health of the patient. In the later stage of the disease, the joint and a plurality of internal organs of the patient are possibly damaged, and the life health of the patient is greatly threatened.

Psoriasis is classified according to its clinical features and is classified into the ordinary type, the articular type, the pustular type and the erythrodermic type, and more than 90% of psoriasis belongs to the ordinary type. Psoriasis vulgaris mostly appears in the knee, elbow, trunk, four limbs, and the like of scalp, and has the characteristic of symmetrical distribution, wherein basic skin damage is mainly plaque of red papule, silvery white scale is attached to the surface of the psoriasis vulgaris, and punctate bleeding phenomenon occurs if the psoriasis vulgaris is scraped. The facial skin damage is drop-shaped infiltrative erythema and rash; scalp skin damage occurs in the form of obvious fascicles; skin lesions in areas with much friction such as the armpit and breast appear as a symptom of decreased scales, exudation and erosion. The skin damage of the joint psoriasis is similar to the common psoriasis, but the joint symptoms can also appear after the skin damage generally occurs. Any joint may be affected and the incidence of joint deformity increases. The clinical manifestations of erythroderma psoriasis are mainly general flushing of the skin and swelling, a large amount of bran-like scales appear on the skin lesion, and general symptoms such as fever may occur. Pustular psoriasis: it is classified as localized or generalized. Firstly, the skin lesions of the localized pustular psoriasis are limited to palms and soles and are symmetrically distributed, and the skin lesions are small pustules on the basis of erythema. Secondly, the generalized pustular psoriasis frequently occurs acutely, and yellowish white superficial aseptic pimples with different sizes are rapidly appeared on normal skin with skin damage of the psoriasis vulgaris or without skin damage, are densely distributed, can be fused and rapidly reach the whole body, and can have the general symptoms of chill, high fever and the like. Some cases have fever, headache, arthralgia and superficial lymph node swelling.

Psoriasis is a common disease in dermatology, the mild patients mainly take external medicines, and the severe patients can select systemic treatment according to the disease condition. The new hair has small skin damage, and the externally applied medicine is the first choice. The oral medicine can be selected from methotrexate, retinoic acid, glucocorticoid, antibiotic, etc. for treatment. The psoriasis has a long course of disease and tends to relapse easily, and the physical and psychological health of patients is seriously harmed.

Through retrieval, no report that the icariin and the madecassic acid are used together for treating the psoriasis exists at present.

Disclosure of Invention

The invention aims to provide a medical application of a pharmaceutical composition, the pharmaceutical composition takes icariin and madecassic acid as main active ingredients, and concretely relates to the application of the pharmaceutical composition in preparing a medicine for treating psoriasis.

Preferably, the psoriasis is selected from psoriasis vulgaris, arthrosis, erythrodermic psoriasis, pustular psoriasis.

The inventor is dedicated to the development of treatment medicines for a long time, and unexpectedly finds that the combined application of icariin and madecassic acid has good treatment effect on psoriasis on the basis of the previous research, and the combination of the two medicines has obvious synergistic treatment effect.

The embodiment 1 of the invention shows that the composition of icariin and madecassic acid has obvious therapeutic effect on psoriasis mice, can obviously improve the histomorphological change of psoriasis models, improves the infiltration number of skin lesion inflammatory cells and the longitudinal epidermal area value, and has obvious therapeutic effect on psoriasis.

Example 2 of the invention shows that the composition of icariin and madecassic acid has obvious therapeutic effect on psoriasis mice by external application, and can relieve psoriasis symptoms.

The effect of the pharmaceutical composition for treating psoriasis is not only obviously superior to the effect of two drugs when used separately, but also superior to the addition of the two effects; the two medicines are combined for use, so that the respective dosage can be obviously reduced, and adverse reactions are reduced; meanwhile, the clinical medication selection of patients with psoriasis is increased, and the medicine has good clinical application prospect.

The asiatic acid in the pharmaceutical composition is one of asiatic acid, salt thereof or a complex thereof.

The weight ratio of the icariin to the madecassic acid in the pharmaceutical composition is preferably 1: 0.01-30.

The pharmaceutical composition also comprises a pharmaceutically acceptable carrier or auxiliary material.

The invention also provides a pharmaceutical preparation containing the pharmaceutical composition, and the pharmaceutical composition can be prepared into a proper and clinically acceptable pharmaceutical preparation according to the needs.

Preferably, the clinically acceptable dosage form is an oral preparation, an injection preparation or an external preparation.

Preferably, the oral formulation is one or more of a tablet, a capsule or a microemulsion formulation thereof.

The inventor considers the requirement of convenient medication for patients and the characteristic that two medicaments are convenient to take at one time compared with the two medicaments taken in a plurality of times, and prepares the medicinal composition into solid medicinal preparations, such as tablets, capsules, granules, pills, dropping pills and the like according to the properties of the active ingredients of madecassic acid and icariin. Wherein the tablet comprises common tablet, coated tablet, sugar coated tablet, film coated tablet, enteric coated tablet, effervescent tablet, chewable tablet, multilayer tablet, disintegrating tablet, dispersible tablet, sublingual tablet, buccal tablet, sustained release tablet, etc. The invention adopts the solid pharmaceutical preparation, has the advantages of convenient carrying and use and simple and easy administration route, and is easy to be accepted by patients.

Preferably, the injection preparation is one or more of injection liquid or injection microemulsion thereof.

Preferably, the external preparation is one or more of powder, ointment, gel, cream, spray or patch, and the administration mode of the external preparation is skin administration.

The pharmaceutical composition can be clinically used for treating psoriasis. In the medical application, the medicinal composition of icariin and madecassic acid can be prepared into a proper medicinal preparation according to the condition of animals and the application part so as to be convenient to use, and the administration time and the administration frequency of the medicinal composition for treating psoriasis need to be determined according to the specific diagnosis result of the condition of the animals, which is within the technical range mastered by the technicians in the field. For example, it will be apparent to one of ordinary skill in the art that a therapeutic regimen for psoriasis in mice is applied to humans and that the effective dose of all drugs to humans can be converted to the effective dose of the drug to mice.

Compared with the prior art, the pharmaceutical composition has the following outstanding advantages in the aspect of treating psoriasis:

(1) compared with the single administration of the icariin or the madecassic acid, the pharmaceutical composition containing the icariin and the madecassic acid has better treatment effect on psoriasis models, and the two pharmaceutical active ingredients show good synergistic effect in the aspect of treating psoriasis.

(2) The pharmaceutical composition contains madecassic acid and icariin, and the combined use of the madecassic acid and the icariin has a synergistic effect and can obviously reduce the use amount of the madecassic acid and the icariin, thereby obviously reducing the toxic and side effects of patients during medication and reducing the occurrence of adverse reactions.

(3) The pharmaceutical composition treats psoriasis in a fixed combination mode, is more convenient to take compared with the mode of taking single medicine simultaneously, improves the compliance and compliance of patients, and enhances the treatment effect on the disease to a certain extent.

Detailed Description

The present invention will be further described below by way of specific embodiments, but the scope of application of the present invention is not limited to the following examples. Alterations and/or combinations of features of the invention will be apparent to those skilled in the art from the disclosure, spirit and/or scope of the invention, and are intended to be encompassed by the invention.

Example 1 Effect of the composition on Imquimod-induced psoriasis-like skin lesions in mice

1. Animal modeling, grouping and administration

1.1 Molding and grouping

The BABL/c mice were depilated on their backs, approximately 2cm by 1.5cm in area, and gently maneuvered to avoid skin damage. The method comprises dividing 70 mice into 7 groups by random number table method, feeding 10 mice each group in cages, and feeding with sterilized water and common mouse feed every day.

Randomly selecting a group as a normal control group, and smearing distilled water on the hairless area of the back about 0.5mL every day; and (3) coating 5% imiquimod cream on the back hairless areas of the other 6 groups, wherein the amount of the imiquimod cream is 60mg/d (the effective drug is 3mg/d), and the number of the continuous 6d areas is less than or equal to the number of the red spot scaling skin damage on the back of the mouse, and judging that the model is successfully prepared.

60 mice successfully molded are divided into a model control group, an icariin group, a madecassic acid group, a composition A group, a composition B group and a composition C group, wherein each group comprises 10 mice.

After the model is successfully prepared, different medicines are respectively given according to groups for treatment.

1.2 administration of drugs

Icariin group: 1mg/kg icariin;

madecassic acid group: 20mg/kg madecassic acid;

composition group A: 10mg/kg icariin +2mg/kg madecassic acid;

composition B group: 1mg/kg of icaritin +20mg/kg of madecassic acid;

composition group C: 10mg/kg icariin +1mg/kg madecassic acid;

each test drug was suspended in 0.5% sodium carboxymethylcellulose and administered 1 time a day for 2 weeks. The normal control group and the model control group were given equal volume of 0.5% sodium carboxymethylcellulose 1 time per day.

In the administration process, except for the normal control group, 5% of imiquimod cream (60mg/d) is applied to the hairless area on the back of the mice of the other groups every other day so as to reduce skin damage of the model mice and naturally relieve the interference on the experiment.

2. Experimental methods and data processing

2.1 Experimental animal specimen Collection

After the experiment, the mice were sacrificed by cervical dislocation. Skin biopsy was performed to collect the back skin lesion tissue, which was fixed in 10% formaldehyde solution for paraffin section preparation and HE staining.

2.2 Observation of the erythematous scaling lesion area and severity of each group of mice

The vital signs of the mice are carefully observed in the treatment process, and the change condition of the erythema scales at the skin lesion is observed at the same time every day.

2.3HE staining method for detecting histological changes in skin

And (3) preparing an HE stained section of the tissue at the skin lesion of the mouse, and observing the infiltration number of inflammatory cells and the change of the thickness of the epidermis. Observing each section under a microscope multiplied by 200, and counting inflammatory cell infiltration number (number/HP) after amplifying by adopting image processing software; and simultaneously, drawing the area of the longitudinal epidermis by using image processing software and taking the substrate membrane band as a boundary, and converting the area into a numerical value, wherein the larger the value is, the thicker the skin damage thickness is.

2.4 data statistics and analysis

Data toShown, analysis of variance was performed using SPSS17.0 software.

3. Results and discussion

3.1 Observation of the area and severity of erythrosquamous lesions in mice

After 2 weeks of treatment, the normal control group had normal back skin and hair growth.

Compared with the normal control group, the skin lesions on the back of the model control group have obvious erythema and scale formation, and the skin is obviously thickened and is similar to the skin lesions of patients with psoriasis.

Symptoms of each treatment group were improved over the model control group, with the composition showing a significant reduction in erythema scales in each group. The reduction in erythema scales was more pronounced in composition A and composition B compared to composition C.

3.2 histological changes in the skin of groups of mice after 2 weeks of treatment

HE staining shows that compared with a normal control group, the skin lesion epidermal acanthocyte layer of the model control group is obviously thickened, the horny layer hyperkeratosis, parakeratosis, dermal papilla vasodilatation and tortuosity, the dermal layer and the subcutaneous tissue inflammatory cell infiltration are obvious, and the histological performance of the skin lesion is similar to that of the psoriasis patient.

Compared with the model control group, the skin lesion thickness of the other drug treatment groups is obviously reduced, the epidermis layer is smoother, and inflammatory cell infiltration is also obviously reduced, wherein the skin lesion thickness of each group of the composition is more obviously reduced, and the inflammatory cells of the composition A group and the composition B group are most obviously reduced. The number of inflammatory cell infiltrates and the longitudinal epidermal area values for each group are shown in Table 1.

TABLE 1 skin lesion inflammatory cell infiltration and longitudinal epidermal area values for each group of mice

Compared with the model control group,^#P<0.05，^##P<0.01; compared with the icariin group,^&P<0.05，^&&P<0.01; compared with the madecassic acid group,^★P<0.05，^★★P<0.01; compared with the composition C group,^＊P<0.05，^＊＊P<0.01。

the combination of the icaritin and the madecassic acid has obvious synergistic effect compared with a single medicine, and has obvious treatment effect on psoriasis. This is of great significance in clinical application.

EXAMPLE 2 Effect of topical composition on Imquimod-induced psoriasis-like lesions in mice

1. Animal modeling, grouping and administration

1.1 Molding and grouping

BABL/c mice were shaved at the ears, approximately 2cm by 1.5cm in area, and gently maneuvered to avoid skin damage. The method comprises dividing 70 mice into 7 groups by random number table method, feeding 10 mice each group in cages, and feeding with sterilized water and common mouse feed every day.

Randomly selecting a group as a normal control group, and smearing distilled water on the hairless area of the ear about 0.5mL every day; the remaining 6 groups were continuously applied with 250mg:12.5mg of imiquimod cream once per morning to the ear for 7 days to prepare an imiquimod-induced psoriasis mouse model. Each administration group continuously and externally applies drugs with different concentrations to the ear skin of the mice in a modeling process. 60 mice successfully molded are divided into a model control group, an icariin group, a madecassic acid group, a composition A group, a composition B group and a composition C group, wherein each group comprises 10 mice.

The mouse models of each group are obtained after erythema scales appear locally.

1.2 administration of drugs

Different treatments were given according to group:

icariin group: icaritin 10 ug/time;

madecassic acid group: madecassic acid 2 mg/time;

composition group A: icaritin 100 ug/time + madecassic acid 200 ug/time;

composition B group: icaritin 10 ug/time + madecassic acid 2 mg/time;

composition group C: icaritin 100 ug/time + madecassic acid 100 ug/time;

each test drug was dissolved in diethylene glycol monoethyl ether and administered 1 time a day for 2 weeks. The normal control group and the model control group were given equal volume of diethylene glycol monoethyl ether 1 time per day.

2. Experimental methods and data processing

Comparative analysis of psoriasis-like mouse model for external treatment: the mice were observed for gross changes, comparing the extent of erythema and scaling. And (3) dehydrating the collected mouse skin lesion gradient alcohol, performing HE staining after dimethylbenzene is transparent, measuring the thickness of the mouse ear epidermis by using a vernier caliper, and comparing the inflammatory cell infiltration degree.

3. Results and discussion

3.1 comparison of symptoms in groups of mice

The mice are continuously externally applied with imiquimod cream for 7d to model psoriasis, and the ear skin of the mice is observed to turn red and enlarged, the surface of the mouse is attached with scale, and the blood vessels are expanded obviously. In the modeling process, various groups of medicines with different concentrations are continuously and externally applied to the skin of the mouse, the phenotype of the mouse is observed after 7 days, and compared with a model control group, the erythema scale of the mouse is obviously reduced and the epidermis thickness is obviously reduced after the medicines of various groups are externally applied.

3.2 comparison of ear edge epidermis thickness in groups of mice

The ear rim thickness was measured and found to be improved in each treatment group compared to the model control group, with the improvement in ear rim thickness being more evident in each group of the composition. The ear rim thickness was more pronounced in composition a and composition B groups compared to composition C group. See table 2.

TABLE 2 ear edge thickness variation for each group of mice

Group of	n	Ear edge thickness (mm)
			Normal control group	10	0.22±0.027##
Model control group	10	0.69±0.071
			Icariin group	10	0.53±0.064#
Madecassic acid group	10	0.49±0.051#
			Composition group A	10	0.35±0.037##&&★★＊
Composition group B	10	0.31±0.032##&&★★＊
			Composition group C	10	0.42±0.056##&★

Compared with the model control group,^#P<0.05，^##P<0.01; compared with the icariin group,^&P<0.05，^&&P<0.01；

compared with the madecassic acid group,^★P<0.05，^★★P<0.01; compared with the composition C group,^＊P<0.05，^＊＊P<0.01。

3.3 comparison of pathological examination of skin tissues of mice in each group

The ears were subjected to a pathological examination of the skin tissue, and HE staining showed: compared with a blank control group, the ear parts of the mice in the model control group have parakeratosis and hyperkeratosis, the thickening of the epidermis acanthosis is obvious, the dermis superficial layer blood vessels expand, inflammatory cells infiltrate, the symptoms of the psoriasis are obviously relieved by each treatment group, and the number of the dermal infiltration lymphocytes is obviously reduced. Wherein the thickness of the skin lesion of each group of the composition is reduced more obviously, and the inflammatory cells of the group A and the group B of the composition are reduced most obviously.

The results suggest that the external icariin, madecassic acid and the composition group can directly act on keratinocytes to relieve psoriasis symptoms, and the treatment effect of each group of the composition is obviously enhanced, so that the composition has better psoriasis treatment effect.

EXAMPLE 3 preparation of tablets

Prescription: 10g of icaritin, 2g of madecassic acid, 25g of microcrystalline cellulose, 20g of lactose and 1.5g of magnesium stearate.

Mixing the above materials and adjuvants uniformly, granulating by conventional wet method, drying, and tabletting.

EXAMPLE 4 preparation of injection

Prescription: 10g of icaritin, 20g of madecassic acid, 800g of sodium chloride and a proper amount of water for injection.

Dissolving sodium chloride in water for injection under stirring, adding icariin and madecassic acid, dissolving, adding water for injection to total amount, filtering, bottling, and sterilizing.

EXAMPLE 5 preparation of capsules

Prescription: 10g of icaritin, 2g of madecassic acid, 30g of starch, 10g of microcrystalline cellulose, a proper amount of 2% hydroxypropyl cellulose (HPMC) aqueous solution and 0.5g of magnesium stearate.

Mixing icariin, madecassic acid, starch and microcrystalline cellulose, adding 2% HPMC water solution, and stirring to obtain soft material. Granulating, oven drying, adding magnesium stearate, grading, mixing, and making into capsule.

EXAMPLE 6 preparation of the ointment

Prescription: 0.15g of icaritin, 0.25g of madecassic acid, 5.45g of PEG4005.45g, PEG33504.00g and 0.30g of hexadecanol.

Dissolving anhydroicaritin, madecassic acid, hexadecanol, PEG400 and PEG3350 in water bath under heating, stirring, and condensing.