CN111388467A

CN111388467A - New application of indigo

Info

Publication number: CN111388467A
Application number: CN202010274779.1A
Authority: CN
Inventors: 刘靖; 莫念; 赖家文; 张思文; 杨丽红; 陈健勤; 成雪莉; 张粤琳
Original assignee: First Affiliated Hospital of Guangzhou University of Chinese Medicine
Current assignee: First Affiliated Hospital of Guangzhou University of Chinese Medicine
Priority date: 2020-04-09
Filing date: 2020-04-09
Publication date: 2020-07-10

Abstract

The invention relates to a new application of indigo, in particular to an application of indigo in preparing a medicament with a psoriasis treatment effect. Innovative research shows that the indigo has excellent treatment effect on psoriasis, can achieve treatment effect by using a small amount of medicine, has good curative effect, can be used for preparing medicines for preventing, relieving and treating psoriasis, provides a new medicine selection for effectively treating psoriasis, and has good application and development prospects. Meanwhile, the raw material resources of the indigo are wide, and the prepared medicine is low in cost and has practical application value.

Description

New application of indigo

Technical Field

The present invention relates to a novel use of compounds, in particular indigo.

Background

Psoriasis is an abnormal immunity chronic proliferative skin disease which is genetically determined by multiple genes and induced by multi-environmental factor stimulation, and is one of dermatological difficult diseases. The disease usually attacks repeatedly, is lingering and difficult to cure, and can cause skin damage to the whole body when the skin damage is serious, accompanied by a large amount of desquamation and severe pruritus, thereby bringing serious influence on physical and mental health of patients. At present, for the traditional Chinese medicine treatment of psoriasis, no externally applied ointment prepared from simple, effective and cheap traditional Chinese medicine monomers exists.

For example, photochemotherapy (UVA, UVB) is a standard treatment mode for psoriasis skin diseases, but requires a corresponding treatment center and has potential photosensitivity and carcinogenicity, local external treatment is usually applied to psoriasis patients with skin damage range of less than 5%, local treatment alone or combined with phototherapy may be insufficient for moderate to severe psoriasis patients, systemic treatment is applied when patients are resistant to local treatment, the skin damage range is too large or the life quality of the patients is reduced, liver and kidney damage and the like can occur in long-term application, and in recent years, various high-efficiency biological agents can be used for treating moderate and severe psoriasis, but the long-term application of TNF- α inhibitor can increase the risk of tumor.

Therefore, the development of a medicament with good psoriasis treatment effect, high safety and low cost is an urgent problem to be solved.

Disclosure of Invention

Based on this, there is a need to provide a new application of indigo. The new application refers to the application of indigo in preparing medicines with the effect of treating psoriasis.

The specific technical scheme is as follows:

application of indigo in preparing medicine for treating psoriasis is disclosed.

In one embodiment, the psoriasis is psoriasis vulgaris.

In one embodiment, the indigo is effective in treating psoriasis by inhibiting hyperkeratosis or parakeratosis of epidermal keratinocytes.

In one embodiment, the indigo is effective in treating psoriasis by inhibiting acanthosis hypertrophy.

In one embodiment, the medicament comprises indigo and a pharmaceutically acceptable excipient.

In one embodiment, the dosage form of the drug is an ointment.

In one embodiment, the medicament comprises indigo blue and white petrolatum.

In one embodiment, the weight percentage of the indigo in the medicine is 0.1-1%.

In one embodiment, the preparation method of the medicine comprises the following steps:

dissolving the indigo blue by a solvent, and mixing and grinding the indigo blue and the white vaseline.

In one embodiment, the solvent is Dimethyl sulfoxide (DMSO) in an amount of 2 to 7 μ L per mg of the indigo.

Compared with the prior art, the invention has the following beneficial effects:

according to the invention, the research shows that the indigo has an excellent treatment effect on psoriasis, the treatment effect can be achieved by using a small amount of medicine, the curative effect is good, the indigo can be used for preparing medicines for preventing, relieving and treating psoriasis, a new medicine selection is provided for effectively treating psoriasis, and the indigo has good application and development prospects. Meanwhile, the raw material resources of the indigo are wide, and the prepared medicine is low in cost and has practical application value.

Drawings

FIG. 1 shows the skin lesions of mice in the blank, model and positive control groups, as well as high and low concentrations of indigo;

FIG. 2 shows pathological sections of mice in high and low concentrations of indigo, and in the blank, model and positive control groups.

Detailed Description

The novel use of indigo according to the invention is described in more detail below with reference to specific examples. The present invention may be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Indigo naturalis also called indigo naturalis, indigo naturalis powder, indigo naturalis flower, blue dew, lake flower, and indigo foam flower, and is dried pigment of plants such as Dicotyledoneae plant, such as fructus Isatidis, Dicotyledoneae plant, such as Caesalpinia japonica, Dicotyledoneae plant, such as Caesalpinia tinctoria, or Polygonaceae plant, such as folium Polygonum tinctorii. Regarding the efficacy application of indigo naturalis, the Chinese pharmacopoeia records that:

1. the anti-tumor comprises 200mg/kg of indigo naturalis component indirubin which is intraperitoneally injected or subcutaneously injected for 7 consecutive days, the composition has an inhibiting effect on a rat cancer W256 solid tumor, 100mg/kg of indigo naturalis component indirubin has an inhibiting effect on mouse sarcoma-180 (S180), 50mg/kg of indirubin is intragastrically infused for 10 consecutive days, the composition has an inhibiting effect on mouse L ews lung cancer, the indirubin treats chronic myelocytic leukemia, bone marrow white cells are rapidly reduced, a large number of degenerative necrotic cells are observed by an electron microscope, juvenile granulocytes are both performed in a swelling and soluble necrosis mode, cytoplasm and nucleus are both obviously damaged, 200mg/kg of indirubin is intraperitoneally injected for 3-5 consecutive days, the inhibiting rate of (3H-TdR) on rat W256 tumor tissue deoxyribonic acid (DNA) is 46%, but the inhibiting effect of 3H-uracil riboside (3H-UR) on tumor tissue doped with ribonucleic acid (RNA) and 3H-phenylalanine, and the in vitro experiment shows that 3 × 10 of the 3H-T-G-DNA is lower than that in vitro experiment^-6The concentration of mol/L can inhibit the 3H-TdR from being mixed into cancer cells of rat W256, mouse Ehrlich Ascites Carcinoma (EAC) and mouse liver cancer (HepA), has little inhibition effect on regenerative liver cells, has no influence on the nucleic acid and protein synthesis of mouse lymphocyte leukemia (L1210) cells, and the concentration of 4 mu g/m L of indirubin has the inhibition effect on the TdR being mixed into human chronic granulocytic leukemia (chronic granulocytic) cellsTherefore, the incorporation of acute granule TdR is more strongly inhibited, which shows that indirubin has greater inhibiting and killing effects on proliferating cells with more active DNA synthesis, but has no inhibiting effect on the incorporation of TdR into naive lymphocytes.

2. Effect on leukemia cells: pharmacological research preliminarily shows that indirubin has the function of destroying leukemia cells. From the aspect of ultrastructural morphology, under the action of indirubin, the cells of degeneration necrosis are mostly in swelling and lytic necrosis. In the experiment, the indirubin is found to enhance the phagocytic capacity of the mononuclear macrophagy system of animals. The monocyte macrophage system plays a certain role in the immune response of the organism, so that the anticancer effect of indirubin is probably related to the improvement of the immune capability of the organism.

3. Bacteriostasis: the tryptanthrin is an active component for resisting dermatophytosis fungi through antifungal tests, and has strong inhibiting effects on microsporum lanosum, tinea circinata, microsporum gypseum, trichophyton purpureum, trichophyton gypseum, trichophyton rubrum, epidermophyton floccosum and the like.

Therefore, in the prior art, the efficacy research aiming at the indigo naturalis mainly focuses on the effective component indirubin. The other component, indigo (also known as food blue No. 1, food cyan No. 2, food blue, acid indigo and hardened indigo), is a disodium salt of 3,3 ' -dioxo-2, 2 ' -biindolyl-5, 5 ' -disulfonic acid, and is a water-soluble non-azo colorant widely used in the food and textile printing and dyeing industries under the general cognition of people. In addition, some studies have disclosed that indirubin in indigo naturalis has a certain therapeutic effect on psoriasis, and in these studies, indigo is generally treated as an impurity, the content of which is controlled to be 0 or close to 0 as much as possible.

The invention discovers that the indigo has excellent treatment effect on psoriasis through innovative research, can achieve treatment effect by using a small amount of medicine, has good curative effect, can be used for preparing medicines for preventing, relieving and treating psoriasis, provides a new medicine selection for effectively treating psoriasis, and has good application and development prospects. Meanwhile, the raw material resources of the indigo are wide, and the prepared medicine is low in cost and has practical application value. The specific technical scheme is as follows:

the embodiment of the invention provides application of indigo in preparing a medicament with the effect of treating psoriasis.

In one particular embodiment, the psoriasis is psoriasis vulgaris.

Psoriasis can be classified into psoriasis vulgaris, pustular psoriasis, joint psoriasis and erythrodermic psoriasis. Among them, psoriasis vulgaris is common and manifested as three signs of psoriasis, namely, wax dripping, punctate bleeding and pellicle. Psoriasis vulgaris lesions first develop in the scalp and the hair appears as bunched hairs that appear as a cloudy, punctate depression in the nail plate and deformation of the nail plate. The indigo provided by the embodiment of the invention is particularly suitable for treating psoriasis vulgaris.

In one specific embodiment, the indigo is effective in treating psoriasis by inhibiting hyperkeratosis or parakeratosis of epidermal keratinocytes.

In one specific embodiment, the indigo is effective in treating psoriasis by inhibiting acanthosis hypertrophy.

In one specific embodiment, the medicament comprises indigo and a pharmaceutically acceptable excipient.

In one embodiment, the pharmaceutical formulation is an ointment.

In one specific embodiment, the drug comprises indigo blue and white petrolatum.

In one specific embodiment, the weight percentage of the indigo in the medicine is 0.1-1%.

In one specific embodiment, the preparation method of the medicine comprises the following steps:

In one specific embodiment, the solvent is Dimethyl sulfoxide (DMSO) in an amount of 2 to 7 μ L per mg of the indigo.

In addition, the embodiment of the invention also provides a medicine with the effect of treating psoriasis, which comprises indigo and pharmaceutically acceptable auxiliary materials.

In one particular embodiment, the psoriasis is psoriasis vulgaris.

Psoriasis can be classified into psoriasis vulgaris, pustular psoriasis, joint psoriasis and erythrodermic psoriasis. Among them, psoriasis vulgaris is common and manifested as three signs of psoriasis, namely, wax dripping, punctate bleeding and pellicle. Psoriasis vulgaris lesions first develop in the scalp and the hair appears as bunched hairs that appear as a cloudy, punctate depression in the nail plate and deformation of the nail plate. The medicine provided by the embodiment of the invention is particularly suitable for treating psoriasis vulgaris.

In one embodiment, the pharmaceutical formulation is an ointment.

Additionally, embodiments of the present invention also provide a method of treating psoriasis comprising the step of applying a medicament comprising indigo to the affected area.

In one particular embodiment, the psoriasis is psoriasis vulgaris.

Psoriasis can be classified into psoriasis vulgaris, pustular psoriasis, joint psoriasis and erythrodermic psoriasis. Among them, psoriasis vulgaris is common and manifested as three signs of psoriasis, namely, wax dripping, punctate bleeding and pellicle. Psoriasis vulgaris lesions first develop in the scalp and the hair appears as bunched hairs that appear as a cloudy, punctate depression in the nail plate and deformation of the nail plate. The methods provided by the embodiments of the present invention are particularly useful in the treatment of psoriasis vulgaris.

In one embodiment, the pharmaceutical formulation is an ointment.

The following is a study of the clinical efficacy of the specific indigo ointment.

Preparation of indigo ointment

1. Indigo monomer: purchased from Doctory Elfin Biotechnology Ltd, under product batch No. D-035-181216, wherein the purity of indigo is > 99%.

2. The materials comprise 10m L syringe, 50m L centrifuge tubes (5 tubes), 0.6m L EP tubes (4 tubes), white vaseline, monomer, electronic scale (accurate to 0.001g), electric grinder, grinding rod, and small melon spoon.

3. The preparation method of the indigo ointment comprises the following steps of firstly, taking a 0.6m L EP tube and balancing on an electronic scale, then respectively weighing 10mg and 100mg of indigo monomers, respectively dissolving the indigo monomers in DMSO (50 mu L), placing the indigo monomers aside for standby, taking a marked 50m L centrifuge tube and balancing, pulling out a core of a 10m L injector, adding a proper amount of white vaseline into the injector, then placing the core again, respectively injecting the white vaseline into the bottom of the centrifuge tube and weighing until the amount of the white vaseline is equal to the total weight of the amount of the indigo monomers to be added into the corresponding centrifuge tube and the total weight of the white vaseline is 10g, pouring the dissolved indigo monomers into the corresponding centrifuge tube after the vaseline is weighed, grinding the indigo ointment by using a grinding rod, and fully grinding the indigo ointment with the weight percentage of 0.1% and 1% respectively.

(II) therapeutic study on psoriasis

1. Test drugs and drugs

5% imiquimod (imported, idale, available for purchase in dermatology hospitals in Guangdong province);

positive control drug: 999 compound dexamethasone acetate ointment;

treatment groups: 1% indigo ointment, 0.1% indigo ointment;

the administration mode comprises the following steps: the administration is carried out on the skin.

2. Laboratory animal

The BA L B/C mice were purchased from animal laboratories of Guangzhou university of traditional Chinese medicine, and were fed to SPF-grade laboratory animal houses of the first subsidiary hospital of Guangzhou university of traditional Chinese medicine.

3. Test methods and results

3.1 Experimental methods

40 male mice of SPF grade BA L B/C, 6-7 weeks old, weighing 17-21g, were randomly grouped into 8 mice per group, wherein,

preparing skin and vaseline without medicine, scraping hair on the back of the mouse to expose a rectangular hairless area of about 3cm × 4cm on the back of the mouse, and uniformly coating the vaseline on the hairless area on the back of the mouse in the blank group by using a sterile medical cotton swab, wherein the thickness is about 1-2mm, and the vaseline is applied once in the morning and evening;

preparing skin, modeling and vaseline without medicine, wherein the modeling method comprises scraping hair on the back of a mouse to expose a rectangular hairless area of about 3cm × 4cm on the back of the mouse, uniformly coating an imiquimod ointment on the hairless area on the back of the mouse in the imiquimod model group by using a sterile medical cotton swab, wherein the thickness of the ointment is about 1-2mm, the ointment is applied once a day, and then uniformly coating the vaseline on the back of the mouse by using the sterile medical cotton swab once in the morning and at night;

dexamethasone control group (positive control group): preparing skin, molding and dexamethasone, and uniformly smearing dexamethasone ointment on a hairless area on the back of a mouse in a dexamethasone control drug group by using a sterile medical cotton swab while molding, wherein the thickness is about 1-2mm, and the ointment is applied once in the morning and at night;

high concentration treatment group (1% indigo ointment group): preparing skin, molding and 1% indigo ointment, and uniformly coating the 1% indigo ointment on the hairless area on the back of a high-concentration treatment group mouse by using a sterile medical cotton swab while molding, wherein the thickness is about 1-2mm, and the ointment is applied once in the morning and at night;

low concentration treatment group (0.1% indigo ointment group): skin preparation, modeling and 0.1% indigo ointment, and when modeling is carried out as described above, the 0.1% indigo ointment is uniformly smeared on the hairless area on the back of the mice of a low-concentration treatment group by using a sterile medical cotton swab, the thickness is about 1-2mm, and the ointment is applied once in the morning and at night.

All groups of mice were fed with feed and drinking water at a constant temperature of 25 ℃ for 7 days. Meanwhile, general conditions of each group of mice were observed and recorded on days 3, 5, and 7, respectively, and recorded and evaluated according to a scoring system. On day 7, each group of mice was sacrificed by cervical dislocation and immediately the skin of the target lesion was cut with a sterile surgical scissors and sectioned to prepare pathological sections.

3.3 results of the experiment

Seven days after administration, the mice in each group were in good condition and active, and the skin lesions of the mice in each group are shown in fig. 1. After 7 days of administration, the pathological tissue sections of the local skin lesions were examined under a 400-fold microscope, and the pathological sections are shown in FIG. 2.

As can be seen from FIG. 1, the skin damage of the shaved area on the back of the blank group is not abnormal, and part of the hair grows rapidly; the epidermises of the imiquimod model group appear obvious erythema scales on the third day of administration, the change of the erythema scales is aggravated along with the experiment, and partial skin lesions fall off; the thickening and hardening conditions of the surface skin of the dexamethasone control group are obvious compared with those of the blank group, but the dexamethasone control group is lighter than that of the imiquimod model group, and no shedding-like change is seen; the mouse erythema scales and epidermis thickening and hardening of the 0.1% indigo ointment group and the 1% indigo ointment group are more obvious than those of the blank group and are lighter than those of the imiquimod model group. The 0.1% indigo ointment group and the 1% indigo ointment group have inhibiting effect on psoriasis skin lesions of mice modeled by imiquimod, including erythema, scale, epidermal thickening, etc.

As can be seen from fig. 2, no psoriasis-like skin lesion manifestations such as hyperkeratosis, parakeratosis, acanthosis thickening and epidermal downward edge are seen in the blank group, psoriasis-like pathological images such as hyperkeratosis, parakeratosis and acanthosis thickening are seen in the imiquimod model group, and the thickness of the epidermis of the mice in the dexamethasone control group, the 0.1% indigo ointment group and the 1% indigo ointment group is lighter than that of the imiquimod group and thicker than that of the mice in the blank group. The 0.1% indigo ointment group and the 1% indigo ointment group have inhibitory effects on hyperkeratosis, parakeratosis and acanthosis hypertrophy in psoriasis lesions of mice modeled by imiquimod.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. Application of indigo in preparing medicine for treating psoriasis is disclosed.

2. Use according to claim 1, wherein the psoriasis is psoriasis vulgaris.

3. The use according to claim 1, wherein the indigo has therapeutic efficacy in the treatment of psoriasis by inhibiting hyperkeratosis or parakeratosis of epidermal keratinocytes.

4. The use according to claim 1, wherein the indigo is effective in treating psoriasis by inhibiting acanthosis hypertrophy.

5. The use according to any one of claims 1 to 4, wherein the medicament comprises indigo and a pharmaceutically acceptable excipient.

6. The use of claim 5, wherein the medicament is in the form of an ointment.

7. The use of claim 6, wherein the medicament comprises indigo blue and white petrolatum.

8. The use according to claim 7, wherein the indigo is present in the medicament in an amount of 0.1 to 1% by weight.

9. Use according to claim 7 or 8, characterized in that the process for the preparation of the medicament comprises the following steps:

10. The use according to claim 9, wherein the solvent is dimethyl sulfoxide in an amount of 2-7 μ L per mg of indigo.