Compounds targeting blood vessels and their medical devices and their application in phototherapy for reducing redness

Abstract

A compound, as shown in Formula I, is a multi-heterocyclic compound. It has been verified that the compound provided by this invention integrates a novel compound targeting the calcitonin gene-related peptide (CGRP) based on ephedrine. Upon contact with the skin, due to photoactivated cell biological function regulation and the blocking effect of the multi-heterocyclic compound I structure on serotonin receptors, it achieves therapeutic effects on cutaneous vascular malformations and telangiectasia, improves the effectiveness of phototherapy in improving chronic cutaneous vascular diseases, and promotes skin wound healing.

Description

Technical Field

[0001] This invention relates to a compound, and more particularly to a compound that has a targeted effect on vascular nerve regulation, which can close proliferating and abnormally dilated blood vessels through phototherapy, and its application in the treatment of skin wounds. Background Technology

[0002] Skin lesions characterized by capillary malformations and telangiectasias are among the most common chronic skin diseases in clinical practice, including port-wine stains (also known as nevus flammeus), acne, rosacea, and infected wounds. These diseases occur in all age groups and can affect both men and women. The main symptoms are erythema and flushing of the skin; in cases with secondary infections, inflammatory papules and pustules may appear, and in severe cases, proliferative nodules and scars may remain, affecting appearance. Current treatments have limited effectiveness for chronic erythema and flushing, as well as recurrent and persistent infected lesions; many patients suffer significant psychological distress due to the noticeable changes in appearance caused by these symptoms. Therefore, there is an urgent clinical need for a more effective treatment for erythema, flushing, and inflammatory wounds that are difficult to heal.

[0003] Currently, phototherapy (including low-energy light, intense pulsed light, and pulsed dye lasers) is one of the most promising treatment methods for "redness reduction" and photodynamic sterilization. Under light irradiation in the 400–1200 nm wavelength range, the photobiological regulation of the skin modulates the microenvironment of vascular endothelial cells and improves cellular energy metabolism, thereby inhibiting angiogenesis; light activates intracellular ROS substances for sterilization, improves excessive local inflammatory responses in refractory wounds, and promotes wound repair. However, current topical medications and phototherapy are slow to control refractory capillary diseases and are not ideal for controlling acne inflammation caused by Propionibacterium acnes infection, exhibiting adverse reactions such as skin irritation and photosensitivity. While existing low-energy phototherapy can alleviate symptoms through repeated treatments, it still suffers from unstable efficacy and a high recurrence rate, and repeated blue light irradiation can easily lead to skin pigmentation. Currently, the gels used in phototherapy are mainly ultrasound imaging gels to improve the penetration and uniformity of light in skin tissue, but no gel itself can currently improve symptoms through targeted neuromodulation effects on blood vessels. Summary of the Invention

[0004] One object of the present invention is to provide a compound that is targeted to blood vessels and achieves a neuromodulatory effect in regulating angiogenesis without causing direct damage to blood vessels.

[0005] Another objective of this invention is to provide a compound that enhances the contractile effect on dilated capillaries and improves the efficiency of phototherapy in improving vasodilation.

[0006] One object of the present invention is to provide the use of a compound in the preparation of a medicament for treating skin diseases (topical).

[0007] Another object of the present invention is to provide the use of a compound in the preparation of a medical device for phototherapy.

[0008] A compound comprising a multi-heterocyclic compound I as shown in Formula I.

[0009]

[0010] The compounds of the present invention further include a group R, which is covalently linked to compound I and ephedrine or an analogue thereof. Group R is an organic compound selected from ester, ether, amide, C1-C10 hydrocarbon groups, or one or more polymers.

[0011] Another compound is shown in Formula II.

[0012]

[0013] Verification has shown that the compound provided by this invention is a novel compound that integrates calcitonin gene-related peptide (CGRP) targeting ephedrine. Upon contact with the skin, due to the photoactivated regulation of cell biological functions and the blocking effect of the multi-heterocyclic compound I structure on serotonin receptors, it achieves therapeutic effects on vascular malformations and telangiectasia of the skin, improves the effectiveness of phototherapy in improving chronic vascular skin diseases (such as erythema, flushing, and sensitivity, including port-wine stains, hemangiomas, acne, rosacea, hypertrophic scars, keloids, and chronic wounds), and promotes skin wound healing.

[0014] The compounds of this invention can be used to prepare drugs and medicated medical devices (such as medicated gels, sprays, creams and dressings) for phototherapy of various types of vascular skin diseases, achieving a highly effective and safe redness reduction effect. Attached Figure Description

[0015] Figure 1 A schematic diagram of the synthetic route for preparing one embodiment of the novel compound of the present invention;

[0016] Figure 2 The above is an NMR result diagram of the novel compound obtained in this invention;

[0017] Figure 3 This is a skin redness-reducing effect diagram of the novel compound prepared according to an embodiment of the present invention;

[0018] Figure 4 This is a graph showing the quantitative analysis results of the novel compound of an embodiment of the present invention on the distribution of blood vessels;

[0019] Figure 5 This is a statistical chart showing the level of inflammatory factor secretion in vascular endothelial cells inhibited by a novel compound according to an embodiment of the present invention.

[0020] Figure 6 Photographs showing the skin redness reduction effect of a novel compound according to an embodiment of the present invention on combined phototherapy;

[0021] Figure 7 This is a graph showing the quantitative analysis results of the effect of a novel compound combined with phototherapy on vascular distribution according to an embodiment of the present invention;

[0022] Figure 8 This is a TUNEL staining result of the effect of a novel compound combined with phototherapy on apoptosis of skin tissue cells according to an embodiment of the present invention.

[0023] Figure 9 This is a comparison chart of the phototherapy effects of a novel compound medical device and a control medical device according to an embodiment of the present invention; wherein, A is a photo of the redness reduction effect of each experimental group before phototherapy, 1 day after phototherapy, and 3 days after phototherapy, and B is a statistical result chart of each experimental group before phototherapy, 1 day after phototherapy, and 3 days after phototherapy. Detailed Implementation

[0024] The technical solution of the present invention is described in detail below. The embodiments of the present invention are only used to illustrate the technical solution of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solution of the invention without departing from the spirit and scope of the technical solution of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention.

[0025] In the following examples, unless otherwise specified, all starting materials and reagents were commercially available from sources such as Selleck, Sigma-Aldrich, and Thermo Fisher Scientific. The obtained materials and reagents were used directly without further purification.

[0026] 1 The 1H NMR spectra were obtained using a Bruker instrument (400 MHz), and chemical shifts are expressed in ppm. Tetramethylsilane was used as an internal reference standard (i.e., 0.00 ppm). 1 H NMR representation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublets. If the coupling constant is provided, its unit is Hz.

[0027] The preparation methods provided in this invention should be understood as examples provided for the purpose of sufficient disclosure, and not as limitations on the scope of protection claimed by this invention. Those skilled in the art can prepare the various compounds provided by this invention by following the guidance of textbooks, lab manuals, or the examples listed in this invention. The preparation of these compounds is also within the general skills of those skilled in the art and can be accomplished under the guidance of existing technology. Specifically, the preparation methods of some compounds provided by this invention are as follows: Figure 1 As shown.

[0028] In the following embodiments of this invention, one-way ANOVA and post-hoc Tukey's test were used for comparisons among multiple groups. For paired comparisons, a two-tailed Student's t-test was used. GraphPad Prism software was used to analyze and plot the data. Each experiment was performed in triplicate, and the results are expressed as mean ± standard error (SEM). p < 0.05 indicates that the difference between groups is statistically significant. "*" indicates p < 0.05, and "**" indicates p < 0.01.

[0029] Example 1: Preparation of the compound

[0030] The method for synthesizing the compounds of this invention is as follows:

[0031] (1) Synthesis of 2-bromo-1-phenylprop-1-one. NBS and silica gel were added to a methanolic solution of acetone. The reaction mixture was stirred at 80 °C for 5 hours. The mixture was filtered, and the filtrate was concentrated under vacuum. The residue was diluted with water, extracted with EA, and the combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under vacuum to give the title product as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 8.07–7.99 (m, 2H), 7.60–7.54 (m, 1H), 7.53–7.48 (m, 2Hs), 5.30 (q, J = 6.6 Hz, 1H), 1.91 (d, J = 6.6 Hz, 3H).

[0032] (2) Synthesis of 1-(1-(1-oxo-1-phenylprop-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one. TEA and 2-bromo-1-phenylprop-1-one were added to a MeOH solution of 1-(piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one, and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated under vacuum. The residue was diluted with water, extracted with EA, and the combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE:EA = 2:1) to give the title compound as a yellow solid. 1H NMR (400MHz, CDCl3) δ 8.67 (s, 1H), 8.12 (d, J = 7.7Hz, 2H), 7.60 (t, J = 7.3Hz, 1H), 3H), 1.87-1.81 (m, 1H), 1.80-1.70 (m, 1Hs), 1.33 (d, J = 6.7Hz, 3H).

[0033] (3) Synthesis of 1-(1-(1-hydroxy-1-phenylprop-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one. NaBH4 was added to a MeOH solution of 1-(1-(1-oxo-1-phenylprop-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one, and the mixture was stirred at room temperature for 2 hours. The mixture was quenched with saturated NH4Cl, extracted with EA, and the combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (PE:EA = 2:1) to give the title compound as a yellow solid. ESI-MS (M+H)+: 352.2.

[0034] (4) Synthesis of 1-(1-(1-hydroxy-1-phenylprop-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one. 1-(1-(1-hydroxy-1-phenylprop-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one was purified by SFC to give a white solid compound, denoted as: Novel Compound.

[0035] The obtained novel compound was analyzed by NMR (see Figure 2The results were obtained by ¹H NMR (400MHz, DMSO-d6) and MS identification, with the following specific data: δ 10.84 (s, ¹H), 7.45–7.24 (m, 6H), 7.04–6.95 (m, 3H), 5.00 (s, ¹H), 4.33 (d, J = 9.1Hz, ¹H), 4.18 (s, ¹H), 2.99 (d, J = 10.2Hz, ¹H), 2.84 (s, ¹H), 2.66 (d, J = 9.3Hz, 2H), 2.35 (dd, J = 25.9, 10.9Hz, 2H), 1.69 (d, J = 10.0Hz, 2H), 0.71 (d, J = 6.5Hz, 3H). ESI-MS (M+H)+: 352.3.

[0036] Example 2: Novel compound applied topically to the skin to reduce redness.

[0037] The specific plan is as follows:

[0038] (1) Phorbol-12-myristate-13-acetate (PMA) was dissolved in anhydrous ethanol, and then sterile corn oil was added to make the volume ratio of anhydrous ethanol to corn oil 1:9 and the concentration of PMA 50 ng / ml. After anesthetizing mice with isoflurane, 100 μl of PMA solution was evenly applied to the skin of the mouse ear (once a day for 7 consecutive days) to induce a chronic inflammatory response of the skin, and significant dilation of the blood vessels in the ear was observed.

[0039] (2) The compound obtained in Example 1 was diluted to 1 mg / ml with phosphate buffer at pH 7.4 and added to glycerol to make a final concentration of 1.0%, that is, 0.01 mg of the novel compound per 1 ml of glycerol;

[0040] (3) Apply the novel compound glycerin solution to the reddened area of ​​the skin, incubate in the dark for 20 minutes, then clean it off. Repeat once a day for two consecutive days. Figure 3 As shown, inflammatory edema and telangiectasia of the skin were significantly improved. The skin telangiectasia index was significantly inhibited (see [reference]). Figure 4 ).

[0041] Example 3: Inhibitory effect of novel compounds on vascular cell inflammatory response

[0042] The specific plan is as follows:

[0043] (1) Human vascular endothelial cells (e.g., human umbilical vein endothelial cells HUVEC) cultured in vitro were treated with tumor necrosis factor α (TNFα) at ​​a final concentration of 20 ng / ml for 24 h to induce inflammatory response in HUVEC cells.

[0044] (2) Remove the cell culture medium containing TNFα;

[0045] (3) Add DMEM complete cell culture medium containing a novel compound at a final concentration of 5 μM to HUVEC cells cultured in vitro and incubate overnight in the dark.

[0046] (4) Total RNA was extracted from lysed HUVEC cells and reverse transcribed. The changes in the transcription levels of granulocyte-macrophage colony-stimulating factor (GMCSF) and interleukin-6 (IL6) genes secreted by human umbilical vein endothelial cells were detected by real-time quantitative PCR (Real-time qPCR).

[0047] (5) Collect HUVEC cell culture supernatant and use enzyme-linked immunosorbent assay (ELISA) to detect changes in the levels of GMCSF and IL6 secreted by human umbilical vein endothelial cells.

[0048] like Figure 5 As shown, the relative mRNA expression levels of pro-inflammatory cytokines (IL-6 and GM-CSF) in vascular endothelial cells, as well as the synthesis and secretion of their corresponding proteins, were inhibited by the novel compound, which was statistically significant.

[0049] Example 4: Novel compound combined with phototherapy to reduce redness

[0050] The specific plan is as follows:

[0051] (1) Dilute the novel compound to 5 mg / ml with phosphate buffer at pH 7.4 and add it to medical coupling agent (medical gel) to make the final concentration of the novel compound 5 mg / 100g gel, i.e. 0.05%;

[0052] (2) Apply the medical coupling agent containing the novel compound to the flushed area of ​​the skin, and perform one session of low-energy phototherapy at a wavelength of 500–1200 nm (energy density 10 J / cm²). 2 After intervention, skin inflammation (redness, swelling) significantly improved (see [link]). Figure 6 Furthermore, the proliferation and dilation of skin capillaries were significantly inhibited (see [reference]). Figure 7 ).

[0053] Example 5: Novel compound combined with phototherapy inhibits macrophage infiltration

[0054] The specific plan is as follows:

[0055] (1) Prepare skin for mice after anesthetizing them with isoflurane;

[0056] (2) Apply 50 μL of a 0.05% solution of the novel compound to the skin of mice, incubate in the dark for 10 minutes, and then clean it off. Repeat once a day for two consecutive days; or apply a medical gel containing 0.05% of the novel compound to the skin of mice, followed by IPL irradiation with 500 nm to 1200 nm light (energy density 10 J / cm²) once. 2 );

[0057] (3) Seven days after the intervention, the mice were sacrificed, and skin tissue samples were taken from the intervention area, fixed, and immunofluorescence staining (DAPI and F4 / 80 double staining) was performed to detect and analyze the infiltration level of macrophages and other inflammatory cells in the skin tissue.

[0058] like Figure 8 As shown, both topical application of the novel compound and the combination of the novel compound with phototherapy can inhibit PMA-induced infiltration of inflammatory cells, including macrophages, in skin tissue, and the effect of the novel compound combined with phototherapy is better.

[0059] Example 6: Novel Compound Medical Device Combined with Phototherapy Control Experiment

[0060] Compared to commonly used phototherapy gel medical devices (ultrasound coupling agents) in clinical practice, the novel compound in this application is more effective and stable in treating chronic skin inflammation and vasodilation, such as... Figure 9 As shown, when applying the same dose of phototherapy, the novel compound group exhibited a significantly lower number of dilated capillaries compared to existing gel medical devices. This demonstrates that, compared to currently used clinical phototherapy gel medical devices (i.e., ultrasound coupling agents), the novel compound is more effective and stable in treating chronic skin inflammation and vasodilation.

Claims

1. A compound, characterized in that... The structural formula is as follows: 。 2. Use of the compound according to claim 1 in the preparation of a medicament for treating a skin disease, which is a vascular malformation or a telangiectatic disease.

3. Use of the compound according to claim 1 in the preparation of a medicament for treating a skin disease, which is a vascular malformation or a telangiectatic disease.

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