WO2022021981A1 - Di-(benzimidazole)-1, 2, 3-triazole derivative and preparation therefor and use thereof - Google Patents

Abstract

The present invention belongs to the technical field of medicinal micro-molecules. Specifically disclosed are a brand-new di-(benzimidazole)-1, 2, 3-triazole derivative, preparation therefor and use thereof and a preparation containing the active ingredient. Researches of the present invention find that the brand-new compound has an unexpected effect in the aspect of inhibiting miR-210 expression, and can be used for developing medicines for pathological miR-210 overexpression related diseases.

Description

Bis-(benzimidazole)-1,2,3-triazole derivatives and their preparation and application technical field

The invention belongs to the technical field of medicine, and in particular relates to a small molecule with medicinal activity.

Background technique

miR-210 is a classic hypoxia-inducible miRNA molecule, and its expression level is regulated by Hypoxia inducible factor-1α (HIF-1α). Studies have confirmed that miR-210 is involved in the regulation of biological processes such as cell proliferation, apoptosis, mitochondrial metabolism, DNA damage repair and angiogenesis. Hypoxia is a common pathological condition that is common in various solid tumors, inflammatory reactions, and ischemic diseases. Current studies have shown that miR-210 is abnormally expressed in many solid tumors, cardiovascular and cerebrovascular diseases, inflammatory diseases, and bone metabolic diseases, and it is particularly closely related to inflammatory and immune responses. CD4 ⁺ T cells, including regulatory T cells and helper T cells, play a very important role in the initiation of immune responses and the maintenance of immune homeostasis. They can provide help to other cells and undertake a variety of effector functions. , when the homeostasis is disrupted, disease states such as autoimmune activation, infection and tumor immunity will arise. TH1 and _TH2 are the classic subtypes involved in immune response; _TH17 is a new cell lineage that secretes inflammatory factors such as IL-17 and IL-22 _. A large number of studies have shown that excessive activation of _TH 17 and _TH 1 is one of the important pathogenesis of many autoimmune diseases, inflammatory diseases and solid tumors. Abnormal pathways play a role in targeted therapy. In recent years, due to the important biological functions of miRNAs and their important roles in various diseases, many techniques for interfering with the expression of miRNAs have been invented. In vivo administration of miRNA mimics, synthetic specific double-stranded small RNA molecules and miRNA overexpression vectors can restore the level of downregulated miRNAs in vivo; while the use of antimiRs, antisense, and antagomiRs can interfere with overexpressed miRNAs. However, there are currently no methods and strategies for the treatment of inflammatory diseases and autoimmune diseases based on miRNAs. Therefore, it is of great significance to develop new small-molecule drugs targeting miRNA with high efficacy, low side effects and low cost.

SUMMARY OF THE INVENTION

The first object of the present invention is to provide a bis-(benzimidazole)-1,2,3-triazole derivative with a new structure.

The second object of the present invention is to provide a method for preparing the bis-(benzimidazole)-1,2,3-triazole derivative.

The third object of the present invention is to provide a use of the bis-(benzimidazole)-1,2,3-triazole derivative for preparing a drug for inhibiting the expression of miR-210.

The fourth object of the present invention is to provide a medicine comprising the bis-(benzimidazole)-1,2,3-triazole derivative.

A bis-(benzimidazole)-1,2,3-triazole derivative, having the structural formula of formula 1;

Formula 1

Said n and m are independently integers from 1 to 6;

Among the R ₁ and R ₂ , at least one substituent is an active substituent, and the active substituent is a hydroxyl group, an alkoxy group, a hydroxyalkyl group or an alkoxyalkyl group.

The present invention provides a compound of formula 1 with a brand-new structure, and it is found that the compound with the brand-new structure can unexpectedly inhibit the expression of miR-210 (microRNA-210) based on the intramolecular action of its intramolecular fragments and groups. For the treatment of pathological miR-210 overexpression-related diseases, further, it can inhibit the differentiation of autoimmune inflammatory _TH 17 and _TH 1 cells and the secretion of related cytokines, promote the differentiation of _TH 2 cells and Secretion of IL-4; unexpected pharmacodynamic activity in autoimmune diseases, inflammatory diseases, infections and tumors mainly mediated by elevated _TH17 and/or _TH1 cells, and can be used in this type of Small molecule drug development for disease.

In the present invention, the n is an integer of 1-6; preferably an integer of 2-4; more preferably 3.

In the present invention, the m is an integer of 1-6; preferably an integer of 2-4; more preferably 3.

The research of the present invention unexpectedly found that in the bis-(benzimidazole) core structure, the 1,2,3-triazole ring was innovatively modified at its end and controlled that the ring contained at least one of the bis-(benzimidazole) Oxygen-active groups, which can unexpectedly increase the inhibitory effect on miR-210 based on intramolecular as well as molecular steric interactions. For example, it can improve the inhibitory effect of autoimmune inflammatory _TH17 and _TH1 cell differentiation and the expression of related cytokines, promote the differentiation of _TH2 cells and the secretion of IL-4, and strengthen the inhibition of keratinocyte proliferation. Effect.

In the present invention, the alkoxy group is C ₁ -C ₆ alkoxy group; for example, methoxy group, ethoxy group, propoxy group and the like.

Preferably, the hydroxyalkyl group is a C ₁ -C ₆ alkyl group with one or more hydroxy substituents. For example, a C ₁ -C ₆ linear or branched alkyl group is modified on the 1,2,3-triazole ring, and a hydroxyl group is substituted with the linear or branched alkyl group. For example, the hydroxyalkyl group is hydroxymethyl, hydroxyethyl and the like.

Preferably, the alkoxyalkyl group is a C ₁ -C ₆ alkyl group with one or more alkoxy substituents. For example, a C ₁ -C ₆ linear or branched alkyl group is modified on a 1,2,3-triazole ring, and an alkoxy group is substituted with the linear or branched alkyl group. For example, the structure of the alkoxyalkyl group is:

The R is a straight-chain or straight-chain alkyl group, and the n1 is an integer from 1 to 10; when n1 is not 1, the alkoxyalkyl group is a polyether substituent.

Preferably, among the R ₁ and R ₂ , one of them is a reactive substituent, and the other substituent is H, the reactive substituent, an alkyl group, a phenyl group, a benzyl group or a halogen.

Preferably, the R ₁ is hydrogen; the R ₂ is a hydroxyalkyl group. The hydroxyalkyl group is preferably hydroxymethyl, 1-hydroxyethyl, 1-hydroxypropyl, 2-hydroxyethyl, 2-hydroxypropyl or 3-hydroxypropyl.

Most preferably, the bis-(benzimidazole)-1,2,3-triazole derivative has the following structural formula (Formula 1-A);

Formula 1-A.

The present invention found that the preferred novel compound (Formula 1-A) has a better inhibitory effect on miR-210, for example, in autoimmune inflammatory _TH17 , TH1 cell differentiation and related cytokine secretion _. Inhibition, as well as promotion of _TH2 cell differentiation and IL-4 secretion, and inhibition of keratinocyte proliferation have unexpected efficacy and less toxic side effects.

The present invention also provides a method for preparing the bis-(benzimidazole)-1,2,3-triazole derivative, which is obtained by subjecting the compound of formula 2 and the compound of formula 3 to a cyclization reaction;

Formula 2

Formula 3

In the present invention, the selection ranges of n, m and substituents in the formula 2 and formula 3 are the same.

Preferably, the solvent for the cyclization reaction is methanol. A reaction aid is added to the cyclization reaction, preferably a Cu(I) source. For example, the Cu(I) source can be obtained based on the reaction between the Cu(II) source and the reducing agent in the reaction system.

In the present invention, the compound of formula 2 is obtained by amidation of the compound of formula 4 and the compound of formula 5;

Formula 4

Formula 5.

The selection range of n and m in Equation 4 and Equation 5 is the same as that in Equation 1.

In the present invention, the amidation reaction can be realized based on existing means.

Preferably, in the present invention, in the amidation reaction process, the compound of formula 4 and an activator are activated in advance, and then reacted with the compound of formula 5. The activator is preferably carbodiimide and 1-hydroxybenzotriazole.

In the present invention, the compound of formula 4 is obtained by cyclization of compounds of formula 6 and formula 7, followed by ester hydrolysis;

Formula 6

Formula 7

The R ₃ is a C ₁ -C ₆ alkyl group, and the selection range of n is the same as in formula 1.

The present invention also provides a bis-(benzimidazole)-1,2,3-triazole derivative and pharmaceutically acceptable salts, solvates, co-crystals, derivatized esters, derivatized amides thereof Use of at least one of the compounds (also referred to as an active ingredient in the present invention) for the manufacture of a medicament for inhibiting the expression of miR-210.

The compound of formula 1 described in the present invention can unexpectedly knock down the expression of miR-210, and can be used for the treatment of pathological miR-210 overexpression related indications.

Preferably, the active ingredient is used to prepare a drug that specifically inhibits the expression of miR-210;

A further preferred application is to prepare a drug that specifically inhibits the expression of miR-210, thereby inhibiting the differentiation of _TH 17 and _TH 1 cells and their cytokine secretion, and promoting the differentiation of _TH 2 cells and the secretion of IL-4.

A further preferred application is for the preparation of a drug for the treatment of autoimmune diseases, inflammatory diseases, infections and tumors mainly mediated by elevated expression of miR-210 and elevated _TH17 and/or _TH1 cells .

In the described application, the autoimmune disease, inflammatory disease, and infection include but are not limited to inflammatory skin diseases (preferably cutaneous lupus erythematosus, psoriasis, parapsoriasis, pityriasis rosea, vitiligo, alopecia areata). at least one of), lichen planus, eczema, urticaria, rheumatoid arthritis, spondyloarthritis, inflammatory bowel disease, multiple sclerosis, type I diabetes, Hashimoto's thyroiditis, human immunodeficiency virus (HIV) infection, hepatitis B virus (HBV) infection, etc.

A preferred application of the present invention is to combine the bis-(benzimidazole)-1,2,3-triazole derivatives and their pharmaceutically acceptable salts, solvates, co-crystals, esters, and amide derivatives into Use of at least one (also referred to as active ingredient in the present invention) for the preparation of a medicament for inflammatory skin diseases. For example, the pharmaceutically acceptable salt of the compound of formula 1 can be hydrochloride, acetate, mesylate, besylate, oxalate, citrate, sulfuric acid of the compound of formula 1 Salts and other pharmaceutically acceptable salts in any form. The solvate of the compound of formula 1 can be, for example, other solvates such as the hydrate of the compound of formula 1. The co-crystal of the compound of formula 1 is a co-crystal formed by the active compound of formula 1 and other pharmaceutical ligands. The pharmaceutical ligands are, for example, polycarboxylic acid ligands. The ester derived from the compound of formula 1 can be a compound modified with an ester group based on the hydroxyl group in the structure, in order to achieve the purpose of prodrug design, or to improve solubility and drug efficacy. The amide compound derived from the compound of formula 1 can be a compound modified by amide group based on the primary amino group and secondary amino group in the structure, in order to achieve the purpose of prodrug design, or improving solubility and drug efficacy. In the present invention, the bis-(benzimidazole)-1,2,3-triazole derivative is used as an active ingredient in combination with the existing pharmaceutical preparation technology to prepare any pharmaceutically acceptable pharmaceutical dosage form; For example, it is used to prepare skin transdermal external preparations, injection preparations, oral preparations and the like. Preferably, the inflammatory skin diseases include but are not limited to psoriasis, parapsoriasis, pityriasis rosea, and cutaneous lupus erythematosus.

In the present invention, the bis-(benzimidazole)-1,2,3-triazole derivatives and their pharmaceutically acceptable salts, solvates, co-crystals, derivatized esters, and derivatized amide compounds are added to At least one of them is used as an active ingredient in combination with the existing pharmaceutical preparation technology to prepare any pharmaceutically acceptable pharmaceutical dosage form; for example, for the preparation of skin transdermal absorption external preparations, injection preparations, oral preparations, etc.

The present invention also provides a medicament comprising a pharmaceutically effective amount of an active ingredient, wherein the active ingredient is the bis-(benzimidazole)-1,2,3-triazole derivative or a pharmaceutically acceptable ingredient thereof. At least one of acceptable salts, solvates, co-crystals, derivatized esters, derivatized amide compounds.

The active ingredient of the medicament of the present invention can be the compound of formula 1 (eg free state), the pharmaceutically acceptable salt of the compound of formula 1, the solvate of the compound of formula 1, the co-crystal of the compound of formula 1, the derivative ester, the derivative at least one of the amides.

The pharmaceutically acceptable salt of the compound of formula 1 can be hydrochloride, acetate, mesylate, besylate, oxalate, citrate, sulfate, etc. of the compound of formula 1 A pharmaceutically acceptable salt in any form.

The solvate of the compound of formula 1 can be, for example, other solvates such as hydrate of the compound of formula 1.

The co-crystal of the compound of formula 1 is a co-crystal formed by the active compound of formula 1 and other pharmaceutical ligands. The pharmaceutical ligands are, for example, polycarboxylic acid ligands.

The ester derived from the compound of formula 1 can be a compound modified with an ester group based on the free hydroxyl group in the structure, in order to achieve the purpose of prodrug design, or to improve solubility and drug efficacy.

The amide compound derived from the compound of formula 1 can be a compound modified by amide group based on the free primary amino group and secondary amino group in the structure, in order to achieve the purpose of prodrug design, or improving solubility and drug efficacy.

In the present invention, the medicament further comprises pharmaceutically acceptable excipients. The adjuvant can be any additive other than the active ingredient that is well known to those skilled in the industry, used to make the active ingredient into a formulation and facilitate the exertion of its medicinal effect, for example, it can be a dispersant, a cosolvent. , colorants, sweeteners, disintegrants, stabilizers, corrosion inhibitors, excipients, sprays, antioxidants, etc.

The medicament of the present invention has any pharmaceutically acceptable dosage form. According to the technical solution of the present invention, the active ingredients described in the present invention can be made into any pharmaceutically acceptable dosage form based on the existing formulation technology, equipment and theory.

As a preference; the dosage form of the drug is external preparation, injection preparation or oral preparation.

Preferably, the external preparation is a transdermal absorption preparation or a transmucosal absorption preparation.

For example, a transdermal absorption preparation contains the active ingredient and any known adjuvants that can achieve its transdermal absorption and enhance its transdermal absorption.

Further preferably, the external preparation is a patch, dressing, external spray, gel, cream, ointment, external lotion, external oil solution, foam or suppository.

In the present invention, the injection preparations are preferably intravenous injection preparations, subcutaneous injection preparations, intradermal injection preparations, and intramuscular injection preparations; more preferably, injection powder preparations or injection solutions.

Preferably, the oral preparation is a tablet, capsule, powder, oral solution or suspension.

The medicament of the present invention is a medicament for inhibiting the expression of miR-210. For example, the medicament of the present invention can be used to knock down pathological miR-210 overexpression, and can be used for the treatment of indications related to miR-210 overexpression in pathological conditions.

Preferably, the drug is a drug that specifically inhibits the expression of miR-210, thereby inhibiting the differentiation of _TH 17 and _TH 1 cells and the secretion of cytokines, and promoting the differentiation of _TH 2 cells and the secretion of IL-4.

Further preferably, the drug is a drug for the treatment of autoimmune diseases, inflammatory diseases, infections and tumors mainly mediated by elevated expression of miR-210 and elevated _TH17 and/or TH1 cells _.

Preferably, the autoimmune disease, inflammatory disease, infection includes but is not limited to inflammatory skin disease (preferably at least one of cutaneous lupus erythematosus, psoriasis, parapsoriasis, pityriasis rosea, vitiligo, alopecia areata) a), lichen planus, eczema, urticaria, rheumatoid arthritis, spondyloarthritis, inflammatory bowel disease, multiple sclerosis, type I diabetes, Hashimoto’s thyroiditis, human immunodeficiency virus (HIV) infection, B At least one of hepatitis virus (HBV) infection. Described inflammatory dermatosis is at least one in cutaneous lupus erythematosus, psoriasis, parapsoriasis, pityriasis rosea, vitiligo, alopecia areata; further preferably including but not limited to psoriasis, parapsoriasis, Pityriasis rosea or cutaneous lupus erythematosus.

In the present invention, the drug, in addition to the formula 1 and its related active ingredients described in the present invention, also includes other types of pharmaceutical active ingredients, preferably, other active ingredients that can be used to inhibit the expression of miR-210 . The other active ingredients that can be used to inhibit the expression of miR-210 are the reported pharmaceutical active ingredients that can be used to inhibit the expression of miR-210, especially those that can be used to specifically inhibit the expression of miR-210.

Preferably; the dosage form of the medicament of the present invention is an external preparation; more preferably, it is a transdermal absorption preparation or a transmucosal absorption preparation. For example, a transdermal absorption preparation contains the active ingredient and any known adjuvants that can achieve its transdermal absorption and enhance its transdermal absorption. Further preferably, the external preparation is a patch, dressing, external spray, gel, cream, ointment, external lotion, external oil solution, foam or suppository. In the present invention, the medicine is a medicine for the treatment of inflammatory skin diseases; further preferably, the inflammatory skin diseases include but are not limited to psoriasis, parapsoriasis, pityriasis rosea, cutaneous lupus erythematosus at least one of them. A preferred medicine of the present invention is a topical preparation such as ointment, gel, liniment, cream, etc. for treating inflammatory skin diseases, preferably an ointment, preferably, it comprises an ointment base and the described ointment base dispersed in the ointment base. Active ingredient. In the present invention, the ointment base can be an existing base, or a base that is prepared based on existing theories and is suitable for use. Preferably, the ointment base is an oily base, an emulsion type base or a water-soluble base; preferably an oily base. Further preferably, the ointment base is at least one of white petrolatum, lanolin, and cetyl alcohol; preferably, it includes white petrolatum and lanolin. The ratio of the white petrolatum and lanolin can be adjusted according to the needs of the paste, preferably 100-300:5-30; more preferably 250-300:5-15. The weight percentage of the active ingredient is 0.001%-1%; preferably 0.008%-0.8%.

beneficial effect

The present invention provides a novel compound, and it is found that the novel compound can inhibit the overexpression of pathological miR-210 and can be used for the treatment of diseases related to overexpression of miR-210.

For example, the novel compounds and their derivatives can inhibit the differentiation of inflammatory _TH17 and TH1 cells and the secretion of related cytokines, promote the differentiation of _TH2 cells and the secretion of IL-4, and inhibit keratinocytes _. Unexpected technical effects in overproliferation. In addition, the brand-new compound of formula 1 described in the present invention not only has good effects in terms of drug efficacy, but also has advantages in terms of toxic and side effects, physical properties, etc. For example, the compound described in the present invention has good solubility and Stability, with lower toxic side effects. The invention has good application prospects in the drug development of autoimmune diseases, inflammatory diseases and infectious diseases.

Description of drawings

One, the accompanying drawing of embodiment 1 and comparative example 1 part: wherein:

Fig. 1-1 is the HPLC chart of the formula 1-A (TD-02) synthesized in Example 1;

Figure 1-2 is the H-NMR chart of formula 1-A (TD-02) synthesized in Example 1;

Figures 1-3 to 1-9 are the H-NMR diagrams of TD-01, TD-03, TD-04, TD-05, TD-06, TD-07, and TD-08 prepared in Comparative Example 1, respectively;

Two, the accompanying drawing of the embodiment 2 part: wherein:

Figure 2-1 Expression of miR-210 in CD4 ⁺ T cells of psoriasis patients before and after treatment with compound 16 and eight small molecule compounds (n=4). ^* P<0.05, ^** P<0.01.

Figure 2-2 The proportion of _TH 17, _TH 1, _TH 2 cell subsets in normal human CD4 ⁺ T cells before and after treatment with different concentrations of compound 16, TD-02, TD-05 and TD-07 (A) and the corresponding Expression levels of cytokine IL-17A, IFN-γ, IL-4 gene mRNA (B), (n=4). ^* P<0.05, ^** P<0.01.

Figures 2-3 Effects of different concentrations of Compound 16, TD-02, TD-05 and TD-07 on the proliferation of keratinocytes (HaCaT cells) (n=5). ^* P<0.05, ^** P<0.01.

3. The accompanying drawings of part 4 of the embodiment

Figure 3-1 Topical application of TD-02 significantly improved imiquimod-induced psoriasis-like skin lesions in mice. (A) Expression of miR-210 in skin lesions of mice in each group; (B, C) gross (B) and pathological (C) changes in inflammatory skin lesions of mice in each group; (D, E) mice in each group Epidermal thickness (D) and inflammatory cell infiltration (E) in skin lesions. ^* P<0.05, ^** P<0.01, ^*** P<0.001

Figure 3-2 Topical application of TD-02 can improve the inflammatory disorder in psoriasis-like skin lesions in mice. (A, B) Flow and statistical graphs of the proportion of _TH 17 and _TH 1 cell infiltration in the skin lesions of mice in each group; (CE) mRNA expressions of il17a, ifng and il4 in the skin lesions of mice in each group. ^* P<0.05, ^** P<0.01, ^*** P<0.001

4. The accompanying drawings of part 5 of the embodiment

Figure 4 (such as 4-1 to 4-12) are respectively the microscopic pictures of organ tissue morphology (the scales are all 50 μm).

detailed description

Example 1: Synthesis of Formula 1-A:

In the present invention, taking the synthesis of formula 1-A as an example, its synthetic route (reaction formula) is as follows:

The synthesis steps are

The solvents DMF and THF used in the reaction were anhydrous treated with CaH ₂ , and EtOH was anhydrous treated with metallic Na. The nuclear magnetic resonance apparatus used was Bruker DPX 400-MHz.

Step (1): Synthesis of 5-p-methylpiperazine-2-nitroaniline (4):

At room temperature, 6.0 g of 5-chloro-2-nitroaniline (2) and 7.5 g of anhydrous K ₂ CO ₃ were weighed into a 100 mL round-bottomed flask, 18 mL of anhydrous DMF was added under nitrogen protection, and then 4.2 g of 4.2 g was added dropwise. -Methylpiperazine (3), the reaction system was heated to 110°C for 12h. After the raw material 2 completely disappeared, the reaction solution was cooled to room temperature, diluted with water (500 mL) and the aqueous phase was extracted with ethyl acetate (500 mL*3), and the obtained organic phase was washed with water (300 mL). The organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain the crude product. 15 mL of methanol was added to the crude product, and the mixture was heated to 60° C. to dissolve all the solids, and then slowly lowered to room temperature. After about 60 min, the solid was no longer precipitated, and 4.9 g of yellow solid compound 4 was obtained by filtration with a yield of 60%.

Step (2): Synthesis of 4-amino-3-nitrobenzylimide ethyl ester (7):

3.0 g of 4-amino-3-nitrobenzonitrile was added to a 500 mL three-necked round-bottomed flask, and then 300 mL of anhydrous EtOH was added to vigorously stir to dissolve the solid. Under the ice bath, HCl gas (concentrated sulfuric acid was dropped into the solid sodium chloride) was continuously fed into the above solution for more than 10h, until no more solid was produced. The stirring was continued overnight, and 4.1 g of yellow powdery solid compound 7 was obtained by suction filtration with a yield of 91%.

Step (3): Synthesis of 4-p-methylpiperazine-1,2-diphenylamine (5):

At room temperature, 4.0 g of compound 4 and 2.0 g of palladium on carbon (10%) were weighed into a 100 mL single-necked round-bottomed flask, evacuated, and anhydrous EtOH (45 mL) was added, stirred vigorously, filled with hydrogen, and reacted overnight. The palladium carbon was removed by suction filtration, and concentrated under reduced pressure to obtain 3.5 g of compound 5 as a pale yellow solid with a yield of 100%. The product must be used immediately and cannot be stored for a long time.

Step (4): Synthesis of Compound 8:

At room temperature, 2.8 g of compound 5 and 3.4 g of compound 7 were weighed into a 250 mL single-neck round-bottomed flask, anhydrous EtOH (40 mL) and AcOH (20 mL) were added, and the mixture was heated to 100° C. and refluxed for 4 h. After the reaction was completed, it was cooled in an ice bath, and concentrated under reduced pressure to remove the solvent to obtain a reddish-brown solid. The solid was dissolved in water (30 mL), concentrated aqueous ammonia (20 mL) was added dropwise under an ice bath, and the solution was allowed to stand overnight. The solid was filtered off, the above solid was dissolved in 20 mL of AcOH/MeOH mixed solvent (AcOH:MeOH=1:9), the insoluble matter was filtered off, and then concentrated under reduced pressure to obtain 3.5 g of compound 8 with a yield of 73%.

Step (5): Synthesis of 4-hydroxybutyric acid (11):

At room temperature, 10.2 g of γ-butyrolactone (10) was weighed into a 250 mL single-necked round-bottomed flask, and then purified water (120 mL) was added to start stirring. Under ice bath, 4.8 g of NaOH solid was slowly added to the above reaction solution, and then the temperature was raised to 100°C and refluxed overnight. Stop heating, cool to room temperature, adjust the pH of the solution to about 5 with dilute hydrochloric acid, extract with ethyl acetate (150 mL*3), and dry the organic phase with anhydrous magnesium sulfate. Concentration under reduced pressure gave 7.9 g of compound 11 in a yield of 65%.

Step (6): Synthesis of benzyl 4-hydroxybutyrate (12):

Under nitrogen protection, 6.2g of compound 11, 2.9g of sodium hydroxide and 0.9g of tetrabutylammonium bromide were sequentially weighed into a 250mL single-neck round-bottom flask, acetone (60mL) was added, and 10.8g of benzyl bromide was slowly added dropwise, and heated. Reflux to 65°C overnight. Stop heating, add 15 mL of water to the above system to quench the reaction, wash with saturated NaHSO ₄ (20 mL), NaHCO ₃ (20 mL), and NaCl (20 mL) solutions successively, and dry the organic phase with anhydrous magnesium sulfate. The solvent was removed under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate=5:1) to obtain 4.5 g of compound 12 as a pale yellow oil with a yield of 40%.

Step (7): Synthesis of Compound 13:

Under nitrogen protection, 3.4 g of compound 12, 4.6 g of triphenylphosphine, 2.2 g of m-hydroxybenzaldehyde and THF (30 mL) were sequentially weighed into a 100 mL single-neck round-bottomed flask, lowered to 0 °C, and 3.6 g of diphenylamine was added dropwise with stirring. Diisopropyl azodicarboxylate (DIAD). After the above reaction solution was continuously stirred for 2 h, water (5 mL) was added to quench the reaction, extracted with ethyl acetate (40 mL*3), and the organic phase was dried over anhydrous magnesium sulfate. Concentrate under reduced pressure to obtain a yellow oily crude product, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate=10:1) to obtain 3.1 g of compound 13 as a bright yellow oil in a yield of 60%.

Step (8): Synthesis of Compound 9:

At room temperature, 1.5 g of compound 8 and 0.8 g of palladium on carbon (10%) were weighed into a 100 mL single-necked round-bottomed flask, and anhydrous EtOH (40 mL) was added after vacuuming, and hydrogen was charged to react overnight. Palladium-carbon was filtered off, and concentrated under reduced pressure to obtain 1.4 g of brick-red compound 9 with a yield of 100%. Compound 9 must be prepared and used immediately and cannot be stored for a long time.

Step (9): Synthesis of Compound 14:

At room temperature, 1.2 g of compound 9 and 1.1 g of compound 13 were weighed into a 500 mL single-neck round-bottom flask, and nitrobenzene (120 mL) was added as a reaction solvent and heated to 145° C. for 24 h. The temperature was lowered to room temperature, and petroleum ether was added to the reaction solution until no more precipitation occurred, about 150 mL. The solid was filtered off and dissolved in methanol, the insolubles were filtered off again, the filtrate was collected, and concentrated under reduced pressure to obtain 1.6 g of compound 14 as a reddish-brown solid in a yield of 70%.

Step (10): Preparation of Compound 15:

At room temperature, 0.8 g of compound 14 and 0.6 g of anhydrous potassium carbonate were sequentially weighed into a 50 mL single-necked round-bottomed flask, and DMF (20 mL) and purified water (20 mL) were used as reaction solvents, and heated to 90 °C for reaction for 2 h. Cool to room temperature, concentrate under reduced pressure to remove the solvent to obtain a yellow solid, dissolve the solid in water, adjust the pH to about 5 with dilute hydrochloric acid, and wash with ethyl acetate (20 mL*3). The aqueous phase was collected and concentrated under reduced pressure to obtain 0.8 g of a crude product of compound 15, with a crude yield exceeding 100%.

Step (11): Synthesis of Compound 16:

At room temperature, 0.4 g of the crude product of compound 15, 0.2 g of carbodiimide, 0.1 g of 1-hydroxybenzotriazole and 10 mL of anhydrous DMF were sequentially weighed into a 50 mL single-neck round-bottomed flask, and 90 mg of triethylamine was added under stirring. React 60min, finally drip 82mg 1-azidopropylamine, react overnight. Concentrated under reduced pressure, purified by silica gel column chromatography (dichloromethane:methanol=5:1), the product was collected, and then separated by preparative chromatography to obtain 50 mg of compound 16 with a yield of 12%.

Step (12): Synthesis of TD-02 (Formula 1-A):

Compound 16 (50mg, 0.08mol, 1.0eq), IM1 (11mg, 0.06mmol, 1.0eq) were dissolved in 1mL MeOH, then copper sulfate pentahydrate (2mg, 0.008mol, 0.1eq), Sodium ascorbate (4.75mg, 0.024 mmol, 0.3 eq) was stirred at room temperature overnight. After the reaction, the reaction solution was separated and purified by preparative thin layer plate (MeOH:NH3H2O=15:1) to obtain 20 mg of pale yellow solid, yield: 34%. The HPLC chart of TD-02 is shown in Figure 1-1; the H-NMR chart is shown in Figure 1-2.

Comparative Example 1:

In a similar manner to Example 1, the 1-azidopropylamine in step (11) is changed to diamine, or IM1 in step (12) is changed; the following comparative compounds are synthesized respectively, and their corresponding H-NMR diagrams are shown in Figure 1- 3 to Figure 1-9;

Example 2:

Use the compounds synthesized in Example 1 and Comparative Example 1 to evaluate the efficacy of the drug:

1) Effects of nine small molecule compounds on the expression of miR-210

The peripheral blood of 4 patients with psoriasis vulgaris was collected, centrifuged by density gradient and used to separate CD4 ⁺ T cells with magnetic beads, and then treated with 2 μM compound 16 and TD-01 to TD-08 small molecule compound solutions and corresponding volumes of solvent, respectively, 48 hours later Cells were collected, total RNA was extracted, and the expression of miR-210 was detected by real-time PCR. The results showed that compared with the solvent-treated control group, the expression of mature miR-210 in CD4 ⁺ T cells of all 4 patients with psoriasis was significantly decreased after compound 16, TD-02, TD-05, and TD-07 treatment. And the inhibitory effect of TD-02 on miR-210 is the strongest among the four compounds, while the inhibitory effect of TD-05 and TD-07 on miR-210 is slightly weaker than that of compound 16; TD-01, TD-03, TD- 04, TD-06 and TD-08 had no significant effect on the expression of mature miR-210 in psoriatic CD4 ⁺ T cells (Fig. 2-1). Therefore, we selected TD-02, TD-05 and TD-07 for further functional screening.

2) Effects of compound 16, TD-02, TD-05 and TD-07 on different subsets of CD4 ⁺ T cells

Normal human peripheral blood was collected, CD4 ⁺ T cells were sorted by magnetic beads, activated with anti-CD3 antibody and anti-CD28 antibody for 24 hours (hrs), and 0 nM, 20 nM, 100 nM, 200 nM, 500 nM and 1 μM compound 16, TD-02 were added, respectively. , TD-05 and TD-07 solutions, and continue to incubate for 48hrs. Cells were collected, and flow cytometry was used to detect the proportion of different CD4 ⁺ T cell subsets ( _TH 1, _TH 2 and _TH 17) in each group, and Real-time PCR was used to detect the corresponding cytokine IFN in cells of each group. - mRNA expression levels of γ, IL-17A and IL-4. The results showed that different concentrations of compound 16 and TD-02 could inhibit the proportion of _TH 17 and _TH 1 cells and the secretion of IL-17A and IFN-γ in CD4 ⁺ T cells, and promoted the proportion of _TH 2 cells and the secretion of IL-4. , and the effect of TD-02 was stronger than that of compound 16 (Figure 2-2A,B), while TD-05 and TD-07 had no significant effect on the proportion of CD4 ⁺ T cells and cytokine secretion in different subsets (Figure 2-2A,B). ).

3) Effects of compound 16, TD-02, TD-05 and TD-07 on keratinocyte proliferation

Human keratinocyte line HaCaT cells were used in this experiment. Before treatment for 12hrs, HaCaT cells were seeded in 96-well plates and cultured in a 37°C, 5% _CO2 incubator for 12h. When the cells grow to 80%-90% confluency, add OnM, 20nM, 100nM, 200nM, 500nM, 1μM Compound 16, TD-02, TD-05 and TD-07 solutions to the keratinocytes, respectively, and continue to culture for 24hrs . 4 hrs before the end of the culture, 10 μL of CCK8 solution was added to each well to continue culturing for 4 hrs. Cell proliferation was detected at a wavelength of 450 nm. The results showed that compound 16 and TD-02 could significantly inhibit the proliferation of HaCaT cells, and the inhibitory effect was in a concentration-dependent manner. Cell proliferation was not significantly affected (Figure 2-3).

This example proves that among the above nine small-molecule compounds, TD-02 shows better effects than other compounds: TD-02 unexpectedly effectively inhibits the expression of mature miR-210, thereby inhibiting the differentiation of _TH 17 and _TH 1 cells And cytokine secretion, promote _TH 2 cell differentiation and IL-4 secretion; at the same time, TD-02 can also significantly inhibit the proliferation of keratinocytes. To further explore the effect of TD-02 on imiquimod-induced psoriasis-like skin inflammation and pathological changes in mice, we conducted the following studies.

Embodiment 3: TD-02 ointment small test recipe technology:

1. Optimum composition and preparation process of blank matrix

1.1 Matrix composition

name	Preferred Composition 1	Preferred Composition 2
White Vaseline	18.0g	19.0g
Lanolin	1.0g	1.0g
cetyl alcohol	1.0g	/

1.2 Preparation process

Weigh the formulation amount of auxiliary materials into a beaker, heat it in a 60°C water bath, stir until it is completely dissolved, withdraw from the water bath, and cool at room temperature to solidify.

1.3 Character investigation

Methods: The samples were placed at room temperature, 40 ℃ constant temperature drying box, the upper room of the refrigerator (2-8 ℃) and the lower room of the refrigerator (-20 ℃) overnight overnight, taken out, and observed the change of properties.

result:

40 ℃ sample: no obvious melting, but the viscosity is significantly lower than the room temperature sample; after 30 minutes at room temperature, there is no obvious difference with the room temperature sample;

2-8℃ sample: no obvious difference from room temperature sample;

-20℃ sample: the hardness increases, the viscosity is significantly higher than that of the room temperature sample, but there is no hard block; after 30min at room temperature, there is no obvious difference with the room temperature sample;

2. Small test formulation composition and preparation process

2.1 Composition of prescription

2.2 Preparation process

①Crush the TD-02 API, pass it through a No. 8 sieve, and set aside;

(2) Weigh the ingredients in the recipe amount into a beaker, heat them in a 60°C water bath, stir until completely dissolved, withdraw from the water bath, and cool at room temperature to solidify to obtain a blank matrix;

③Weigh the prescription amount and sieve the TD-02 API, add it to the blank matrix, stir evenly, and then pack it.

Example 4: Pharmacodynamic study of topical TD-02

Adopt the TD-02 preparation that embodiment 3 obtains, carry out drug effect research:

Imiquimod (IMQ) applied to the back skin of mice can induce a psoriasis-like mouse model, and psoriasis-like pathological changes appear on the back skin of mice. Therefore, the IMQ-induced psoriasis model has been widely recognized for the study of psoriasis pathogenesis and therapeutic drugs. We selected 42 healthy female Balb/c mice aged 6-8 weeks, with 2 × 2 cm of back hair removal, and randomly divided them into 7 groups of 6 mice, one of which was normal control mice without any treatment, and the other 6 The experiment was carried out according to the following methods: ①IMQ group: 80mg of 5% IMQ ointment was applied on the back every morning; ②blank matrix group: 80mg of 5% IMQ ointment was applied on the back every morning, and 72mg of blank matrix ointment was applied on the back every afternoon , for 6 consecutive days; ③ Positive control group: apply 5% IMQ ointment 80 mg on the back every morning, and apply calcipotriol ointment 72 mg on the back every afternoon for 6 consecutive days; ④ 0.008% TD-02 group: daily Apply 5% IMQ ointment 80mg on the back in the morning, and apply 0.008% TD-0272mg on the back every afternoon for 6 consecutive days; ⑤0.024% TD-02 group: apply 5% IMQ ointment 80mg on the back every morning, every afternoon 0.024% TD-0272mg was applied on the back for 6 consecutive days; ⑥0.08% TD-02 group: 80 mg of 5% IMQ ointment was applied on the back every morning, and 0.08% TD-0272 mg was applied on the back every afternoon for 6 consecutive days. Mice in each group were photographed every other day and scored for PASI of psoriatic skin lesions. They were sacrificed on the 7th day, and their back skin lesions were removed, and HE staining was performed to observe the pathological changes. At the same time, RNA was extracted to detect the changes of miR-210 and inflammatory cytokines. , the dermal single cell suspension was separated and flow cytometry was used to detect the proportion of _TH 17 and _TH 1 cells. The results showed that: on the 7th day of modeling, compared with normal mice, the expression of miR-210 in the skin lesions of the IMQ group and the blank matrix group was significantly increased (Fig. 3-1A); The expression in the skin lesions of mice in each TD-02 treatment group was significantly decreased in a concentration-dependent manner (Fig. 3-1A). Topical TD-02 ointment can significantly improve IMQ-induced inflammatory skin lesions in mice (including gross and pathological changes), and the 0.08% TD-02 group was the most significant (Figure 3-1B-E). At the same time, topical TD-02 ointment can significantly inhibit the infiltration of _TH 17 and _TH 1 cells in IMQ-induced inflammatory skin lesions in mice (Figure 3-2A, B), inhibit the expression of IL-17A and IFN-γ mRNA, and promote IL-4 mRNA expression (Figure 3-2C-E).

Example 5: Pharmacokinetic and toxicological studies of TD-02 ointment

1 The basic situation of the variety

The test product in this study is TD-02 ointment (prepared in Example 3), and the main indication is the topical treatment of inflammatory skin diseases. The intended clinical route of administration is skin administration, and the intended clinical dosage should not exceed 100g per week.

2 Summary of pharmacological and toxicological studies

Non-clinical trials of TD-02 ointment carried out toxicological studies: Rodents (SD rats) were used as the experimental system for the toxicological studies of TD-02 ointment to investigate the long-term toxic effects of the test substance after transdermal administration, and the results were obtained in The long-term toxicity test is accompanied by the assessment of plasma and skin exposure to determine the toxic reaction, toxic target organ or target tissue of repeated administration of the test substance.

2.1 Summary of toxicological studies

2.1.1 Overview of toxicology studies

The toxicology study of TD-02 ointment used rodents (SD rats) as the experimental system to investigate the long-term toxic effects of the test substance after administration through the skin, and the drug exposure in plasma and skin was evaluated in the long-term toxicity test. , to determine the toxic reaction, toxic target organ or target tissue of repeated administration of the test substance.

2.1.2 Repeated administration toxicity with toxicokinetics test in rats for 14 consecutive days

SPF SD rats were selected as the experimental system, and a 14-day repeated administration toxicity study was carried out in rats according to the proposed clinical route of TD-02 ointment. 40 male SD rats were selected and randomly divided into 4 groups according to body weight, with 10 animals in each group. TD-02 ointment low-dose damaged skin group; TD-02 ointment medium-dose damaged skin group; TD-02 ointment high-dose damaged skin group (drug content: 0.000%, 0.080%, 0.240%, 0.800%, respectively), each The animals in the group were given 10% body surface area, and the coating dose was about 0.03 g/cm ² , once a day, 7 days a week, for a total of 14 days (2 weeks). In the high-dose group, toxicokinetic blood collection was performed for the first administration, and toxicokinetic blood collection and skin collection were performed for the last administration. /2 etc.)

The results showed that (1) the body weight, hematology, coagulation, blood biochemistry and electrolytes, organ coefficients, and morphology of tissues and organs (observed by naked eye and microscope; see Figure 4-1 to Figure 4-12) in each group were all equal. There was no abnormal change related to the test substance (see the attached table for details); (2) The concentration of TD-02 in plasma and skin was detected by UPLC-MS/MS method. The lower limit of quantification in plasma was 1 mg/L, and the linear range was 1-64 mg/L. The concentration of TD-02 in all samples was lower than the lower limit of quantification. TD-02 was not detected in plasma under the current testing conditions, suggesting a low risk of systemic exposure to the drug. The lower limit of skin quantification was 5mg/L, and the linear range was 5-320mg/L. No TD-02 was detected in the skin of the blank group for 30 minutes. g; 3h skin concentration was 19.82±13.65mg/L, and the content was 79.26±54.62mg/g; 24h skin concentration was lower than the lower limit of quantification. According to the characteristics of local administration of the drug, it is suggested that the drug can penetrate the skin, and the clearance time is relatively fast, and it will not accumulate in the skin within 24 hours.

Conclusion: Under the experimental conditions, SD rats were given TD-02 ointment through the skin for 14 days (2 weeks), no obvious accumulation effect of the test substance in the body, and no obvious toxic reaction.

3 Comprehensive evaluation

The test product in this study is TD-02 ointment, and its main indication is the topical treatment of inflammatory skin diseases. The intended clinical route of administration is skin administration, and the intended clinical dosage should not exceed 100g per week. This study was evaluated by the preclinical preliminary pharmacology and toxicology study of TD-02 ointment, including:

(1) Toxicological studies showed that no obvious toxicity was observed after repeated skin administration of TD-02 ointment for 14 consecutive days, and no accumulation of the test substance was found in the body.

Attached table 1 The effect of TD-02 ointment on animal body weight (unit: g,

)

Attached Table 2 The effect of TD-02 ointment on rat hematological indexes (excluding toxic animals)

Note: Compared with the blank control group, ⁺ P≤0.05, ⁺⁺ P≤0.01.

Attached table 3 The effect of TD-02 ointment on biochemical indexes of rats

Note: Compared with blank control group, ⁺ P≤0.05.

Supplementary Table 4 Effects of TD-02 ointment on electrolytes in rats

Note: Compared with the blank control group, ⁺ P≤0.05, ⁺⁺ P≤0.01.

Attached table 5 The effect of TD-02 ointment on rat blood coagulation index

See Figures 4-1 to 4-12 for the shape of the organs:

Fig. 4-1 Liver of blank matrix damaged skin group (1M02 animal). At the end of administration, HE staining, ×200. Hepatocyte cords were arranged in an orderly manner, no degeneration or necrosis of hepatocytes, and no congestion of hepatic sinusoids.

Fig. 4-2 Kidneys of blank matrix damaged skin group (1M02 animal). At the end of administration, HE staining, ×200. The structure of kidney cortex and medulla was clear, the nephron morphology and structure were normal, and there was no inflammatory cell infiltration in the interstitium.

Figure 4-3 The heart of the blank matrix damaged skin group (1M02 animal). At the end of administration, HE staining, ×200. Myocardial fibers were evenly stained, with clear horizontal stripes, no degeneration, necrosis, interstitial hemorrhage, or infiltration of inflammatory cells.

Fig. 4-4 Lungs of blank matrix damaged skin group (1M02 animal). At the end of administration, HE staining, ×200. The structure of the alveolar wall was normal, there was no inflammatory cell infiltration in the interstitium, and no obvious exudate was found in the alveolar cavity.

Figure 4-5 Spleen of blank matrix damaged skin group (1M02 animal). At the end of administration, HE staining, ×200. The structure of the white pulp and red pulp of the spleen was clear, and there was no congestion or fibrous tissue hyperplasia.

Fig. 4-6 Damaged skin of blank matrix damaged skin group (1M02 animal). At the end of administration, HE staining, ×100. Epidermal parakeratosis, thickened acanthus, blisters and crusts can be seen.

Figure 4-7 Liver of TD-02 ointment high-dose damaged skin group (4M01 animal). At the end of administration, HE staining, ×200. Hepatocyte cords were arranged in an orderly manner, no degeneration or necrosis of hepatocytes, and no congestion of hepatic sinusoids.

Fig. 4-8 Kidneys of damaged skin group with high dose of TD-02 ointment (4M01 animal). At the end of administration, HE staining, ×200. The structure of kidney cortex and medulla was clear, the nephron morphology and structure were normal, and there was no inflammatory cell infiltration in the interstitium.

Figure 4-9 Heart of TD-02 ointment high-dose damaged skin group (4M01 animal). At the end of administration, HE staining, ×200. Myocardial fibers were evenly stained, with clear horizontal stripes, no degeneration, necrosis, interstitial hemorrhage, or infiltration of inflammatory cells.

Fig. 4-10 Lungs of TD-02 ointment high-dose damaged skin group (4M01 animal). At the end of administration, HE staining, ×200. The structure of the alveolar wall was normal, there was no inflammatory cell infiltration in the interstitium, and no obvious exudate was found in the alveolar cavity.

Figure 4-11 Spleen of TD-02 ointment high-dose damaged skin group (4M01 animal). At the end of administration, HE staining, ×200. The structure of the white pulp and red pulp of the spleen was clear, and there was no congestion or fibrous tissue hyperplasia.

Figure 4-12 Damaged skin of TD-02 ointment high-dose damaged skin group (4M01 animal). At the end of administration, HE staining, ×100. Focal ulceration of the skin epidermis, parakeratosis, thickening of the acanthus, and crusts can be seen.

Attached table 6 Statistical table of TD-02 ointment on rat organ coefficient

In conclusion, the compound of formula 1-A described in the present invention has excellent efficacy and lower toxic and side effects, and can be used for drug development of inflammatory skin diseases.

Claims (16)

1. A bis-(benzimidazole)-1,2,3-triazole derivative, characterized in that it has the structural formula of formula 1;

   

   Said n and m are independently integers from 1 to 6;

   Among the R ₁ and R ₂ , at least one substituent is an active substituent, and the active substituent is a hydroxyl group, an alkoxy group, a hydroxyalkyl group or an alkoxyalkyl group.
2. The bis-(benzimidazole)-1,2,3-triazole derivative according to claim 1, wherein the alkoxy group is a C ₁ -C ₆ alkoxy group;

   Preferably, the hydroxyalkyl group is a C ₁ -C ₆ alkyl group with one or more hydroxy substituents;

   Preferably, the alkoxyalkyl group is a C ₁ -C ₆ alkyl group with one or more alkoxy substituents.
3. The bis-(benzimidazole)-1,2,3-triazole derivative of claim 1, wherein one of the R ₁ and R ₂ is an active substituent, and the other Substituents are H, the reactive substituents described, alkyl, phenyl, benzyl or halogen.
4. The bis-(benzimidazole)-1,2,3-triazole derivative according to any one of claims 1 to 3, wherein the R ₁ is hydrogen; the R ₂ is hydroxyl alkyl.
5. The bis-(benzimidazole)-1,2,3-triazole derivative according to claim 1, characterized in that, it has the following structural formula;

   
6. A method for preparing a bis-(benzimidazole)-1,2,3-triazole derivative according to any one of claims 1 to 5, wherein the compound of formula 2 and the compound of formula 3 are combined Obtained by cyclization reaction;

   
7. The method for preparing bis-(benzimidazole)-1,2,3-triazole derivatives according to claim 6, wherein the solvent for the cyclization reaction is methanol;

   A reaction aid is added in the cyclization reaction, which is a Cu(I) source.
8. The method for preparing bis-(benzimidazole)-1,2,3-triazole derivatives according to claim 6, wherein the compound of formula 2 is obtained by amidation of the compound of formula 4 and the compound of formula 5;

   

   Preferably, in the amidation reaction process, the compound of formula 4 is activated under an activator in advance, and then reacts with the compound of formula 5;

   The activator is preferably carbodiimide and 1-hydroxybenzotriazole.
9. The method for preparing bis-(benzimidazole)-1,2,3-triazole derivatives according to claim 8, wherein the compound of formula 4 is cyclized by the compound of formula 6 and formula 7, It is then obtained by ester hydrolysis;

   

   

   The R ₃ is a C ₁ -C ₆ alkyl group.
10. A bis-(benzimidazole)-1,2,3-triazole derivative according to any one of claims 1 to 5 and a pharmaceutically acceptable salt, solvate, co-crystal, derivative ester thereof, Use of at least one of the derivatized amide compounds for the preparation of a medicament for inhibiting the expression of miR-210;

    Preferably, for the preparation of a drug that specifically inhibits the expression of miR-210;

    Further preferably, it is used to prepare a drug that specifically inhibits the expression of miR-210, thereby inhibiting the differentiation of _TH 17 and _TH 1 cells and their cytokine secretion, and promoting the differentiation of _TH 2 cells and the secretion of IL-4;

    Further preferably, it is used for preparing and treating autoimmune diseases, inflammatory diseases, infections and tumor medicines mainly mediated by elevated expression of miR-210 and elevated _TH17 and/or _TH1 cells;

    Preferably, the autoimmune disease, inflammatory disease, infection includes but is not limited to inflammatory skin disease, lichen planus, eczema, urticaria, rheumatoid arthritis, spondyloarthritis, inflammatory bowel disease, multiple sclerosis , Type I diabetes, Hashimoto's thyroiditis, human immunodeficiency virus (HIV) infection, hepatitis B virus (HBV) infection, etc.;

    Preferably, at least one of the bis-(benzimidazole)-1,2,3-triazole derivatives and their pharmaceutically acceptable salts, solvates, co-crystals, derivatized esters, and derivatized amide compounds An application for preparing a medicine for inflammatory skin disease; the inflammatory skin disease is at least one of cutaneous lupus erythematosus, psoriasis, parapsoriasis, pityriasis rosea, vitiligo, and alopecia areata; Further preferred include but not limited to psoriasis, parapsoriasis, pityriasis rosea, cutaneous lupus erythematosus;

    Preferably, a pharmaceutically effective amount of the bis-(benzimidazole)-1,2,3-triazole derivative is combined with a pharmaceutically acceptable adjuvant to prepare any pharmaceutically acceptable preparation; the The formulations described include, but are not limited to, topical formulations, oral formulations or injection formulations.
11. A medicine comprising a pharmaceutically effective amount of an active ingredient, wherein the active ingredient is the bis-(benzimidazole)-1,2,3-triazole according to any one of claims 1 to 5 Derivatives, or at least one of pharmaceutically acceptable salts, solvates, co-crystals, derivatized esters, derivatized amide compounds thereof.
12. The medicine of claim 11, further comprising pharmaceutically acceptable excipients.
13. The medicine according to claim 11, wherein the medicine has any pharmaceutically acceptable dosage form; preferably an external preparation, an injection preparation or an oral preparation;

    Preferably, the external preparations are transdermal absorption preparations or transmucosal absorption preparations; more preferably, patches, dressings, external sprays, gels, creams, ointments, external lotions, external oil solutions, Foams or suppositories;

    Preferably, the injection preparation is preferably an intravenous injection preparation, a subcutaneous injection preparation, an intradermal injection, or an intramuscular injection preparation; more preferably, an injection powder or injection;

    Preferably, the oral preparation is a tablet, capsule, powder, oral solution or suspension.
14. The medicine according to any one of claims 11 to 13, wherein the medicine is a medicine for inhibiting the expression of miR-210;

    Preferably, the drug is a drug that specifically inhibits the expression of miR-210, thereby inhibiting the differentiation of _TH 17 and _TH 1 cells and their cytokine secretion, and promoting the differentiation of _TH 2 cells and the secretion of IL-4;

    Further preferably, the medicine is a medicine for the treatment of autoimmune diseases, inflammatory diseases, infections and tumors mainly mediated by elevated expression of miR-210 and elevated _TH17 and/or _TH1 cells;

    Preferably, the autoimmune disease, inflammatory disease, infection includes but is not limited to inflammatory dermatosis, lichen planus, eczema, urticaria, rheumatoid arthritis, spondyloarthritis, inflammatory bowel disease, multiple sclerosis , at least one of type I diabetes, Hashimoto's thyroiditis, human immunodeficiency virus (HIV) infection, and hepatitis B virus (HBV) infection;

    Preferably, the inflammatory dermatosis is at least one of cutaneous lupus erythematosus, psoriasis, parapsoriasis, pityriasis rosea, vitiligo, and alopecia areata; further preferably including but not limited to psoriasis, parapsoriasis Psoriasis, pityriasis rosea, or cutaneous lupus erythematosus.
15. The medicine according to any one of claims 11 to 14, characterized in that the medicine further comprises other types of active pharmaceutical ingredients, preferably other active ingredients that can be used to inhibit the expression of miR-210.
16. The medicament according to any one of claims 11 to 15, wherein the medicament is a medicament for treating inflammatory skin diseases;

    Preferably, the inflammatory skin disease includes but is not limited to at least one of psoriasis, parapsoriasis, pityriasis rosea, cutaneous lupus erythematosus;

    Preferably, the medicine is an ointment, gel, liniment, cream, etc. for treating inflammatory skin diseases, preferably an ointment, preferably, it comprises an ointment base and the active ingredients dispersed in the ointment base;

    Preferably, the ointment base is an oily base, an emulsion type base or a water-soluble base; preferably an oily base;

    Further preferably, the ointment base is at least one of white vaseline, lanolin, and cetyl alcohol; preferably, it includes white vaseline and lanolin;

    More preferably, the weight percentage of the active ingredient is 0.001% to 1%; preferably 0.008% to 0.8%.

Images