Blood vessel targeting compound, medical instrument thereof and application of compound in phototherapy red removal

Abstract

The invention relates to a compound which is a multi-element heterocyclic compound I as shown in a formula I. Through verification, the compound provided by the invention is a novel compound integrated with targeted calcitonin gene related peptide (CGRP) on the basis of ephedrine. When the phototherapy material is in contact with the skin, due to the photoactivated cell biological function regulation effect and the blocking effect of the multi-element heterocyclic compound I structure on a 5-hydroxytryptamine receptor, the treatment effect on skin vascular malformation and capillary dilatation diseases is achieved, the effective rate of phototherapy for improving chronic skin vascular diseases is improved, and skin wound healing is promoted. # imgabs0 #

Description

Technical Field

[0001] The invention relates to a compound, in particular to a compound with a targeted effect on vascular nerve regulation, which can seal proliferating and abnormally dilated blood vessels through light treatment, and its application in skin wound treatment. Background Art

[0002] Skin lesions with capillary malformations and telangiectasias as pathological manifestations are the most common and chronic skin diseases in clinical practice, including port wine stains (also known as port wine stains), acne, rosacea and infected wounds. Such diseases are distributed in all age groups and can occur in both men and women. They are mainly manifested as erythema and flushing of the skin. Infected patients may develop inflammatory papules and pustules. In severe cases, hyperplastic nodules and scars may remain, affecting the appearance. Existing treatments have limited effects on chronic erythema and flushing, as well as recurrent and persistent infected lesions; many patients suffer great psychological distress due to the obvious changes in appearance caused by the above symptoms. Therefore, a more effective treatment for erythema, flushing and inflammatory non-healing wounds is urgently needed in clinical practice.

[0003] At present, relying on phototherapy (including low-energy light, intense pulsed light and pulsed dye laser, etc.) for "redness removal" and photodynamic sterilization treatment is one of the most promising treatment methods. Under the light of wavelength range of 400-1200nm, the photobiological regulation of light on the skin regulates the microenvironment of vascular endothelial cells and improves cell energy metabolism, thereby inhibiting angiogenesis; light activates intracellular ROS substances to kill bacteria, improves excessive local inflammatory response of difficult-to-heal wounds, and promotes wound repair. However, the current topical drugs and phototherapy have a slow effect on the control of refractory capillary diseases, and the control of acne inflammation caused by Propionibacterium acnes infection is not ideal, and has skin irritation and photosensitivity adverse reactions. Although the existing low-energy phototherapy technology (phototherapy) can relieve symptoms through repeated and multiple treatments, there are still problems of unstable efficacy and easy recurrence, and repeated blue light irradiation treatment is prone to skin pigmentation. The gel currently used for phototherapy is mainly ultrasound imaging gel to improve the penetration and uniformity of light in skin tissue, but there is currently no gel itself that can improve symptoms through the neuromodulatory effect of targeting blood vessels. Summary of the invention

[0004] One object of the present invention is to provide a compound that has targeting properties on blood vessels and achieves a neuromodulatory effect on regulating new blood vessels without causing direct damage to the blood vessels.

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

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

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

[0008] A compound includes a polycyclic heterocyclic compound I as shown in formula I.

[0009]

[0010] The compound of the present invention further comprises a group R, which is covalently linked to the compound I and ephedrine or its analogues. The group R is an organic compound selected from one or more of an ester group, an ether group, an amide group, a C1-C10 hydrocarbon group, or a polymer.

[0011] Another compound is shown in formula II.

[0012]

[0013] It has been verified that the compound provided by the present invention is a novel compound integrating targeted calcitonin gene-related peptide (CGRP) on the basis of ephedrine. When in contact with the skin, due to the light-activated cell biological function regulation effect and the blocking effect of the polycyclic heterocyclic compound I structure on 5-hydroxytryptamine receptors, the therapeutic effect on skin vascular malformations and capillary dilation diseases is achieved, and the efficiency of phototherapy for improving chronic skin vascular diseases (such as erythema, flushing and sensitivity, including port wine stains (also known as port wine stains), hemangiomas, acne, rosacea, hypertrophic scars, keloids and chronic wounds, etc.) is improved, and the healing of skin wounds is promoted.

[0014] The compounds of the present invention are used to prepare drugs and medicated medical devices (such as medicated gels, sprays, creams and dressings, etc.), which are applied to phototherapy of various types of skin vascular diseases to achieve efficient and safe redness removal. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0016] Figure 2 This is the NMR result diagram of the novel compound obtained in the present invention;

[0017] Figure 3 This is a diagram showing the skin redness-removing effect of a novel compound prepared in one embodiment of the present invention;

[0018] Figure 4 This is a diagram showing the quantitative analysis results of the vascular distribution of a novel compound according to an embodiment of the present invention;

[0019] Figure 5 This is a statistical graph 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 This is a photo of the skin redness-removing effect of the novel compound in one embodiment of the present invention combined with phototherapy;

[0021] Figure 7 This is a quantitative analysis result diagram of the effect of a novel compound combined with phototherapy on blood vessel distribution according to an embodiment of the present invention;

[0022] Figure 8 This is a graph showing the results of TUNEL staining detection of the apoptosis of skin tissue cells by the novel compound combined with phototherapy according to one embodiment of the present invention;

[0023] Fig. 9 This is a comparison chart of the phototherapy effects of the new compound medical device of one embodiment of the present invention and the control medical device; wherein A is a photograph of the redness removal 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 DESCRIPTION

[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 rather than to limit it. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the invention can be modified or replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.

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

[0026] 1 H NMR spectra were measured using a Bruker instrument (400 MHz), and chemical shifts were expressed in ppm, with tetramethylsilane being used as an internal reference standard (i.e., 0.00 ppm). 1 H NMR notation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of a doublet. Coupling constants, when given, are given in Hz.

[0027] The preparation methods provided by the present invention should be understood as examples for the purpose of full disclosure, rather than as limitations on the scope of protection claimed by the present invention. A person skilled in the art can prepare the various compounds provided by the present invention according to the guidance of textbooks, experimental manuals or the examples listed in the present invention. The preparation of these compounds is also a general skill that a person of ordinary skill in the art should possess, and can be accomplished under the guidance of the prior art. Specifically, the preparation routes of some of the compounds provided by the present invention are as follows Figure 1 shown.

[0028] In the following examples of the present invention, one-way analysis of variance (ANOVA) and post hoc Tukey test were used for multiple group comparisons. For paired comparisons, a two-tailed Student's t test was used. GraphPad Prism software was used to analyze the data and draw the graphs. Each experiment was repeated three times and the results were expressed as mean ± standard error (SEM). p<0.05 indicates that the difference between the groups is statistically significant. "*" indicates p<0.05, and "**" indicates p<0.01.

[0029] Example 1 Preparation of Compounds

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

[0031] (1) Synthesis of 2-bromo-1-phenylpropan-1-one. NBS and silica gel were added to a methanol solution of acetone. The reaction mixture was stirred at 80°C for 5 hours. The mixture was filtered and the filtrate was concentrated in vacuo. 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 in vacuo 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-phenylpropan-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one. TEA and 2-bromo-1-phenylpropan-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 in vacuo. 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 in vacuo. 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-phenylpropan-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one. To a MeOH solution of 1-(1-(1-oxo-1-phenylpropan-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one was added NaBH4, 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 in vacuo. 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-phenylpropan-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one. 1-(1-(1-hydroxy-1-phenylpropan-2-yl)piperidin-4-yl)-1,3-dihydro-2H-benzo[d]imidazol-2-one was purified by SFC to obtain a white solid compound, which was recorded as a novel compound.

[0035] The obtained novel compound was analyzed by NMR (see Figure 2) and MS identification, the specific data are as follows: 1H NMR (400MHz, DMSO-d6) δ10.84(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 externally to the skin to remove redness

[0037] The specific plan is as follows:

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

[0039] (2) The compound prepared in Example 1 was diluted to 1 mg / ml with a phosphate buffer solution of pH 7.4 and added to glycerol to a final concentration of 1.0%, i.e., 0.01 mg of the novel compound was contained in 1 ml of glycerol;

[0040] (3) Apply the glycerol solution of the new compound to the flushed area of ​​the skin, incubate in the dark for 20 minutes, and then clean it off. Do this once a day for 2 consecutive days. Figure 3 As shown in Figure 2, skin inflammatory edema and capillary dilation were significantly improved. The skin capillary dilation index was significantly inhibited (see Figure 4 ).

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

[0042] The specific plan is as follows:

[0043] (1) Tumor necrosis factor α (TNFα) at ​​a final concentration of 20 ng / ml was used to intervene in human vascular endothelial cells (e.g., human umbilical vein endothelial cells HUVEC) cultured in vitro for 24 h to induce an inflammatory response in HUVEC cells;

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

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

[0046] (4) HUVEC cells were lysed to extract total intracellular RNA. After reverse transcription, real-time fluorescence quantitative PCR (qPCR) was used to detect changes in the transcription levels of granulocyte-macrophage colony-stimulating factor (GMCSF) and interleukin-6 (IL6) genes secreted by human umbilical vein endothelial cells;

[0047] (5) The HUVEC cell culture supernatant was collected and the enzyme-linked immunosorbent assay (ELISA) was used to detect the 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 proinflammatory cytokines (IL-6 and GM-CSF) in endothelial cells and the synthesis and secretion of the corresponding proteins were inhibited by the novel compound, which was statistically significant.

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

[0050] The specific plan is as follows:

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

[0052] (2) Apply the medical coupling agent containing the new compound to the flushed area of ​​the skin, and give low-energy phototherapy with a wavelength of 500 to 1200 nm (energy density 10 J / cm 2 After the intervention, skin inflammation (redness and swelling) was significantly improved (see Figure 6 ), and the proliferation and dilation of skin capillaries were significantly inhibited (see Figure 7 ).

[0053] Example 5: Novel compounds combined with phototherapy inhibit macrophage infiltration

[0054] The specific plan is as follows:

[0055] (1) Prepare the skin of mice after isoflurane anesthesia;

[0056] (2) 50 μL of 0.05% glycerol solution of the novel compound was applied to the mouse skin, incubated in dark for 10 minutes, and then cleaned, once a day for 2 consecutive days; or a medical gel containing 0.05% of the novel compound was applied to the mouse skin, followed by irradiation with 500nm-1200nm IPL light once (energy density 10J / cm 2 );

[0057] (3) Seven days after the intervention, the mice were killed, and the skin tissues in the intervention area were sampled and fixed. Immunofluorescence staining (DAPI and F4 / 80 double staining) was performed to detect and analyze the infiltration levels of macrophages and other inflammatory cells in the skin tissues.

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

[0059] Example 6: Comparative experiment of novel compound medical device combined with phototherapy

[0060] Compared with the phototherapy gel medical devices (ultrasound coupling agents) commonly used in clinical practice, the new compounds in this application are more effective and stable in treating chronic skin inflammation and vasodilation, such as Fig. 9 As shown in the figure, when the same dose of phototherapy is performed, the number of dilated capillaries in the new compound group is significantly lower than that of the existing gel medical device. It can be seen that compared with the phototherapy gel medical device (i.e., ultrasound coupling agent) commonly used in clinical practice, the new compound is more effective and stable in treating chronic skin inflammation and vascular dilation.

Claims

1. A compound comprising a polycyclic heterocyclic compound I as shown in formula I.

2. The compound according to claim 1, characterized in that Also included is a group R, wherein the group R is covalently linked to compound I and ephedrine or its analogs, respectively.

3. The compound according to claim 2, characterized in that The group R is an organic substance, selected from one or more of an ester group, an ether group, an amide group, a C1-C10 hydrocarbon group, or a polymer.

4. The compound according to claim 2, characterized in that As shown in formula II.

5. Use of the compound according to claim 1 in the preparation of medicines for treating skin diseases.

6. The use according to claim 5, characterized in that The skin disease is skin vascular malformation or telangiectasia disease.

7. Use of the compound according to claim 1 in the preparation of a drug for removing redness from the skin.

8. A drug for implementing phototherapy, characterized in that The invention comprises the compound according to any one of claims 1 to 4.

9. A medical device, characterized in that The invention comprises the compound according to any one of claims 1 to 4.

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