阅读说明：本技术 用于选择性消除细菌并消融癌细胞的aie活性光敏剂 (AIE-active photosensitizers for selective eradication of bacteria and ablation of cancer cells ) 是由 唐本忠 李琦瑶 王建国 于 2020-07-24 设计创作，主要内容包括：本主题涉及基于AIE的光敏剂(PS),其可用于在白光照射下选择性杀死癌细胞并消除细菌病原体。PS表现出适度水溶性、明亮发射时的高ROS效率、良好生物相容性、对靶标的高特异性、低的暗毒性、显著的光毒性以及有效的单线态氧(～(1)O-(2))生成。PS示出了聚集诱导的ROS生成(AIROSG)效应。(The present subject matter relates to AIE-based Photosensitizers (PS) that can be used to selectively kill cancer cells and eliminate bacterial pathogens under white light illumination. PS exhibits moderate water solubility, high ROS efficiency at bright emission, good biocompatibility, high specificity to target, low dark toxicity, significant phototoxicity, and potent singlet oxygen: ( 1 O 2 ) And (4) generating. PS shows the aggregation-induced ROS production (aiorg) effect.)

1. A photosensitizer, comprising a compound having the following backbone structure:

wherein R is₁Selected from the group consisting of H and alkyl;

wherein each R₂And R₃Independently selected from the group consisting of H, alkyl, unsaturated alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and diphenylamine; and is

X^-Selected from the group consisting of PF^6-、BF^4-、CH₃COO^-、CF₃COO^-、ClO^4-、F^-、Cl^-、Br^-、I^-、(F₃CSO₂)N^-And PO₄ ^3–Group (d) of (a).

2. The photosensitizer of claim 1, wherein the compound is:

3. a method of generating singlet oxygen comprising illuminating the compound of claim 1 with white light.

4. A method of killing cancer cells, comprising:

incubating a target cancer cell with a compound of claim 1; and

while the compound is in contact with the target cancer cell, white light illumination is applied to the target cancer cell to kill the target cancer cell.

5. The method of claim 4, wherein the cancer cells are incubated with the compound for about 12 hours.

6. The method of claim 4, wherein the target cancer cell is in a living animal.

7. A method of killing a target pathogen comprising:

incubating the target pathogen with the compound of claim 1; and

while the compound is in contact with the target pathogen, white light illumination of the target pathogen is performed, thereby killing the target pathogen.

8. The method of claim 7, wherein the target pathogen comprises at least one of a gram-negative bacterium and a gram-positive bacterium.

9. The method of claim 7, wherein the target pathogen is incubated with the compound for about 15 minutes.

10. The method of claim 7, wherein the target pathogen is in a living animal.

11. A photosensitizer, comprising a compound having the formula:

12. a method of generating singlet oxygen comprising illuminating the compound of claim 11 with white light.

13. A method of killing cancer cells, comprising:

incubating a target cancer cell with the compound according to claim 12; and

while the compound is in contact with the target cancer cell, white light illumination is applied to the target cancer cell, thereby killing the target cancer cell.

14. The method of claim 13, wherein the cancer cells are incubated with the compound for about 12 hours.

15. The method of claim 13, wherein the target cancer cell is in a living animal.

16. A method of killing a target pathogen comprising:

incubating the target pathogen with the compound of claim 12; and

while the compound is in contact with the target pathogen, white light illumination of the target pathogen is performed, thereby killing the target pathogen.

17. The method of claim 16, wherein the target pathogen comprises at least one of a gram-negative bacterium and a gram-positive bacterium.

18. The method of claim 16, wherein the target pathogen is incubated with the compound for about 15 minutes.

19. The method of claim 16, wherein the target pathogen is in a living animal.

Technical Field

The present subject matter relates generally to photosensitizers for photodynamic therapy, and in particular to AIE active photosensitizers for use in eliminating bacteria and killing cancer cells.

Background

Infection with pathogens leads to serious illness and high mortality worldwide. For decades since the first discovery of penicillin in 1928, antibiotics have been used primarily to treat diseases caused by bacteria. However, antibiotics are limited to targets and have common side effects such as diarrhea, nausea, and stomach upset, among others. In addition, the emergence of drug-resistant bacteria poses a serious threat to human health after long-term abuse of antibiotics. Recent studies have shown that about 70% of normal e.coli (e.coli) are resistant after 3 hours of exposure to antibiotics. In addition, the development of new antibiotics has not kept up with the development of drug resistant bacteria. Therefore, the development of alternative antibacterial therapeutic agents is imminent, and such development has important significance for clinical applications.

At the same time, cancer is another major public health problem. Globally, about one sixth of the deaths are due to cancer. Approximately 1,762,450 new cancer cases and approximately 606,880 cancer-related death cases were predicted in the united states in 2019. To date, while various treatment modalities such as surgery, chemotherapy, and radiation therapy have been developed, each of these modalities is associated with risks and/or deleterious side effects. In particular, surgery may be at risk because it is invasive to the human body. In addition, surgery often fails to completely and accurately ablate tumors without damaging healthy tissue. Chemotherapeutic drugs such as cisplatin can cause nephrotoxicity, increasing the risk of infection and drug resistance. On the other hand, patients receiving radiation therapy may experience fatigue and hair loss in the treatment area.

In recent years, photodynamic therapy (PDT) has become a promising approach against pathogenic bacteria and a powerful clinical protocol for cancer treatment. PDT employs Photosensitizers (PS) to treatThe destructive singlet oxygen is generated by endogenous oxygen molecules under illumination (¹O₂) Or other Reactive Oxygen Species (ROS). Thus, combining the advantages of PS and light, PDT exhibits several significant advantages, including non-invasiveness, non-drug resistance, low cytotoxicity, selective targeting, spatiotemporal precision, and synergy with conventional therapeutic modalities. Among the major achievements in the development of PDT, fluorescence-induced PDT (FL-PDT) with the unique advantage of real-time monitoring has attracted increasing attention. PS is the key to FL-PDT and plays a crucial role in the therapeutic effect. However, most conventional organic PSs have the problem of aggregation-induced quenching (ACQ) in biological systems, resulting in undesirably weak emission and poor ROS generation. Fortunately, the discovery of the aggregation-induced emission (AIE) phenomenon provides a convenient solution. Unlike ACQ luminophores, AIE luminophores emit very weakly in dilute solutions. However, due to the restricted mechanisms of intramolecular movement (RIM), they show strong emission and ROS generation in the aggregated state. Therefore, AIE-based PS has a wide potential for development in PDT applications. Although the AIE-based PS research has progressed rapidly in anticancer and antibacterial therapies, there are still a number of issues to be addressed. A general problem is that to achieve selective binding, specific targeting ligands must be introduced into the PS. The method is not only complicated in synthesis, but also high in cost. Another significant challenge is that for most existing PS it is rather difficult to modulate their targets by merely modulating the external conditions without changing their molecular structure.

Therefore, there is a strong need for efficient production without additional modification¹O₂And AIE PS selective for pathogens and cancer cells.

Disclosure of Invention

The present subject matter relates to AIE-based Photosensitizers (PS) that can be used to selectively kill cancer cells and eliminate bacterial pathogens under white light illumination. PS can exhibit moderate water solubility, high ROS efficiency at bright emission, good biocompatibility, high specificity to target, low dark toxicity, significant phototoxicity, and efficient singlet oxygen: (¹O₂) And (4) generating. PS can show that aggregation induces ROS production(AIROSG) effect.

PS can recognize and eliminate different targets under different conditions without changing its structure. For example, when the PS is incubated with the first target for a first period of time, the PS can kill or eliminate the first target under white light illumination. Further, when the PS is incubated with the second target for a second period of time, the PS can kill or eliminate the second target under white light illumination. For example, PS can effectively eliminate drug-resistant e.coli infection under white light illumination, while host tissues are unaffected. By extending the incubation time to 12 hours, PS can kill cancer cells with little damage to normal cells.

In one embodiment, the Photosensitizer (PS) comprises a compound having the following backbone structure formula:

wherein R is₁Selected from the group consisting of H and alkyl;

wherein each R₂And R₃Independently selected from the group consisting of H, alkyl, unsaturated alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and diphenylamine; and is

X^-Selected from the group consisting of PF^6-、BF^4-、CH₃COO^-、CF₃COO^-、ClO^4-、F^-、Cl^-、Br^-、I^-、(F₃CSO₂)N^-And PO₄ ^3–Group (d) of (a).

In one embodiment, the compound is:

in one embodiment, the method of generating singlet oxygen may comprise illuminating the compound with white light.

In one embodiment, a method of killing cancer cells can include incubating target cancer cells with a compound and, while the compound is in contact with the target cancer cells, subjecting the target cancer cells to white light illumination to kill the target cancer cells.

In one embodiment, a method of killing a target pathogen may include incubating the target pathogen with a compound and, while the compound is in contact with a target cancer cell, white light illumination of the target pathogen to kill the target pathogen.

Drawings

Various embodiments will now be described in detail with reference to the accompanying drawings.

FIGS. 1A to 1B show that 1(A) has different water fractions (f)_W) PL spectrum of 4TPA-BQ in DMSO/water mixture; FIG. 1(B) is a graph showing relative emission intensity (I/I)₀) To f_WThe figure (a). Illustration is shown: in solution under irradiation of a hand-held UV lamp (f)_W0%) and aggregate (f)_WFluorescence image of 4TPA-BQ in 70%) state. Excitation wavelength: 380 nm; concentration: 10 μ M.

Figure 2 shows the ultraviolet-visible (UV-vis) spectrum of 4TPA-BQ in DMSO solution.

FIG. 3 shows that at f_wGraph of particle size distribution of 4TPA-BQ aggregates in 70% DMSO/water mixture.

FIGS. 4A to 4D show 4(A) anion-. pi.in the single crystal structure of 4TPA-BQ⁺And F-H interaction; 4(B) dihedral angle; 4(C) intermolecular hydrogen bonds; and 4(D) C-H … pi interaction. aE is defined as pi calculated by a single point using the M062X/6-31+ G (d, p) method based on a single crystal structure⁺Interaction energy with anions.

Fig. 5 shows a crystal stack of 4 TPA-BQ.

FIGS. 6A to 6E show UV-vis spectra of 6(A) the PS-free singlet oxygen indicator N, N '-bis (2, 3-dihydroxypropylanthracenedipropionamide) (N, N' -di (2,3-dihydroxypropyl) -9, 10-anthracenedipropanoamide, DHPA); 6(B) UV-vis spectrum of 4TPA-BQ in solution; 6(C) UV-vis spectrum of Ce 6; 6(D) UV-vis spectrum of Rose Bengal (RB); and 6(E) under white light irradiation at f_w99% 4TPA-BQ in DMSO/water mixture in aggregated stateUV-vis spectroscopy. [ PS ]]＝9×10^-6M，[DHPA]＝4.5×10^-5M。

FIGS. 7A to 7D show UV-vis spectra of 7(A) RB and 4 TPA-BQ; 7(B) UV-vis spectra of DHPA and RB; 7(C) at f_wUV-vis spectrum of 4TPA-BQ under white light irradiation in 99% DMSO/water mixture; and 7(D) decomposition rates of DHPA in 4TPA-BQ and RB, respectively. [ DHPA ]]＝5×[PS]Recording the time interval: for 30 s.

FIGS. 8A to 8B show nanosecond transient absorption (ns-TA) spectra of 8(A) 4TPA-BQ in an aqueous solution saturated with argon; and 8(B) triplet lifetime of 4TPA-BQ in argon saturated aqueous solution. Excitation wavelength: 355 nm.

FIGS. 9A to 9D show graphs of the relative absorbance of DHPA under white light illumination of 9(A) without and with different PS, where A₀And a is the absorbance of DHPA at 378nm before and after white light irradiation, respectively. [ PS ]]＝9×10^-6M，[DHPA]＝4.5×10^-5M; 9(B) decomposition rates of DHPA in 4TPA-BQ and RB, respectively; 9(C) calculated energy level diagrams between singlet and triplet states; and 9(D) the Natural Transition Orbital (NTO) of 4 TPA-BQ.

Fig. 10 shows CLSM images of e.coli (e.coli) and s.epidermidis (s.epidermidis) incubated with 5 μ M4TPA-BQ for 15 min.

FIG. 11 shows CLSM images of COS-7 cells and HLF cells incubated with 5 μ M4TPA-BQ for 15 min.

FIG. 12 shows the morphology of E.coli and S.epidermidis incubated with 5 μ M4TPA-BQ with and without white light illumination. Control group: no treatment is carried out; dark group: treatment with 4TPA-BQ only; PDT group: treated with 4TPA-BQ and white light irradiation.

FIGS. 13A to 13F show the bactericidal efficiency of 13(A)4TPA-BQ on E.coli; 13(B) the bactericidal efficiency of 4TPA-BQ on Staphylococcus epidermidis; 13(C) the bactericidal efficiency of 4TPA-BQ on ampicillin-resistant Escherichia coli; 13(D) photographs of culturing E.coli and Staphylococcus epidermidis on agar plates supplemented with 5. mu.M 4 TPA-BQ; and 13(E) to 13(F) cell viability after incubation of COS-7 and HLF cells with 4TPA-BQ for 15 min.

Figure 14 shows CLSM images of PI stained e.coli and HLF cells and staphylococcus epidermidis and HLF cells after incubation with 5 μ M4TPA-BQ under white light illumination. The dashed line indicates HLF cells.

FIGS. 15A to 15D show the antibacterial activity against ampicillin resistant E.coli infection in vivo. 15(A) show photographs of burn wounds of bacterially infected mice treated with PBS only (control), 4TPA-BQ only (dark), and 4TPA-BQ + white light irradiation (PDT), respectively. 15(B) shows the variation in size of infected wounds with different treatments; 15(C) is a photograph of a plate showing the bacterial load of skin wounds and organs at 100-fold dilution on day 5; and 15(D) shows hematoxylin and eosin staining of infected skin sections on days 5 and 10. B: a blood vessel. H: the hair follicle.

FIGS. 16A to 16B show CLSM images of 16(A) COS-7 cells and 16(B) HeLa cells incubated with 10. mu.M 4TPA-BQ over different staining times.

FIGS. 17A to 17B show CLSM images of 17(A) COS-7 cells and 17(B) HeLa cells incubated with 10 μ M4TPA-BQ, followed by illumination with white light, over different staining times.

Detailed Description

The following definitions are provided to understand the subject matter of the present invention and to construct the appended claims.

Definition of

It should be understood that the drawings described above or below are for illustration purposes only. The drawings are not necessarily to scale, emphasis instead generally being placed upon illustrating the principles of the teachings. The drawings are not intended to limit the scope of the present invention in any way.

Throughout this application, where a composition is described as having, including, or comprising a particular component, or where a method is described as having, including, or comprising a particular method step, it is contemplated that the composition of the present teachings can also consist essentially of, or consist of, the recited component, and that the method of the present teachings can also consist essentially of, or consist of, the recited method step.

In the present application, where an element or component is referred to as being included in and/or selected from a list of recited elements or components, it is to be understood that the element or component can be any one of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components. Moreover, it should be understood that elements and/or features of the described compositions, devices, or methods may be combined in various ways, whether explicit or implicit, without departing from the spirit and scope of the invention.

The use of the terms "including" or "having" should generally be understood as open-ended and non-limiting unless otherwise specifically stated.

The use of the singular in the present invention includes the plural (and vice versa) unless specifically stated otherwise. In addition, where the term "about" is used before a quantitative value, the present teachings also include the specific quantitative value itself, unless specifically stated otherwise. As used herein, unless otherwise specified or inferred, the term "about" means a variation of ± 10% from the nominal value.

It should be understood that the order of steps or order of performing certain actions is immaterial so long as the invention remains operable. Further, two or more steps or actions may be performed simultaneously.

As used herein, "heteroaryl" refers to an aromatic monocyclic ring system containing at least one ring heteroatom selected from the group consisting of oxygen (O), nitrogen (N), sulfur (S), silicon (Si), and selenium (Se), or a polycyclic ring system in which at least one ring present in the ring system is an aromatic ring and contains at least one ring heteroatom. Polycyclic heteroaryl groups include two or more heteroaryl rings fused together and a monocyclic heteroaryl ring fused to one or more aromatic carbocyclic, non-aromatic carbocyclic and/or non-aromatic cycloheteroalkyl rings. The heteroaryl group as a whole may have, for example, 5 to 22 ring atoms and contain 1 to 5 ring heteroatoms (i.e., a 5-to 20-membered heteroaryl group). Heteroaryl groups can be attached to the defined chemical structure at any heteroatom or carbon atom, resulting in a stable structure. Typically, the heteroaryl ring does not contain an O-O, S-S or S-O bond. However, one or more of the N or S atoms in the heteroaryl group can be oxidized (e.g., pyridine N-oxide, thiophene S, S-dioxide). Examples of heteroaryl groups may include, for example, a 5-membered monocyclic ring system or a 6-membered monocyclic ring system and a 5-6 bicyclic ring system as shown below:

wherein T is O, S, NH, N-alkyl, N-aryl, N- (arylalkyl) (e.g., N-benzyl), SiH₂SiH (alkyl), Si (alkyl)₂SiH (arylalkyl), Si (arylalkyl)₂Or Si (alkyl) (arylalkyl). Examples of such heteroaryl rings may include pyrrolyl, furyl, thienyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, isothiazolyl, thiazolyl, thiadiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, indolyl, isoindolyl, benzofuranyl, benzothienyl, quinolinyl, 2-methylquinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, benzotriazolyl, benzimidazolyl, benzothiazolyl, benzisothiazolyl, benzisoxazolyl, benzooxadiazolyl, benzoxazolyl, cinnazinyl, 1H-indazolyl, 2H-indazolyl, indolizinyl, isobenzofuranyl, naphthyridinyl, phthalazinyl, pteridinyl, purinyl, oxazolopyridyl, thiazolopyridyl, imidazopyridyl, furopyridinyl, thienopyridinyl, Pyridopyrimidinyl, pyridopyrazinyl, pyridopyridazinyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, and the like. Other examples of heteroaryl groups include 4,5,6, 7-tetrahydroindolyl, tetrahydroquinolinyl, benzothienopyridyl, benzofuropyridinyl, and the like. In some embodiments, heteroaryl groups may be substituted as described herein.

"halo" or "halogen" as used herein refers to fluoro, chloro, bromo and iodo.

As used herein, "alkyl" refers to a straight or branched chain saturated hydrocarbon group. Examples of alkyl groups can include methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, sec-butyl, tert-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), hexyl, and the like. In various embodiments, the alkyl group can have 1 to 40 carbon atoms (i.e., C1-40 alkyl), such as 1 to 30 carbon atoms (i.e., C1-30 alkyl), 1 to 20 carbon atoms, or 1 to 10 carbon atoms. In some embodiments, alkyl groups may have 1 to 6 carbon atoms, and may be referred to as "lower alkyl". Examples of the lower alkyl group may include methyl, ethyl, propyl (e.g., n-propyl and isopropyl), and butyl (e.g., n-butyl, isobutyl, sec-butyl, tert-butyl). In some embodiments, alkyl groups may be substituted as described herein. An alkyl group is typically not substituted with another alkyl, alkenyl, or alkynyl group.

"cycloalkyl" as used herein refers to a saturated, non-aromatic, monovalent mono-or multicyclic radical having 3 to 10, preferably 3 to 6, carbon atoms. The term may be further exemplified by radicals such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, and adamantyl.

As used herein, "heterocycloalkyl" refers to a monovalent saturated 5-to 9-membered monocyclic or bicyclic ring system containing one, two, or three ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon atoms. In the case of monocyclic heterocycloalkyl, the ring is preferably 5-or 6-membered, and in the case of bicyclic heterocycloalkyl, the bicyclic ring is preferably 7-, 8-or 9-membered. As used herein, "heterocycloalkyl" is unsubstituted or substituted. Examples of substituents on heterocycloalkyl may be independently selected from alkyl, hydroxy, hydroxyalkyl, benzyl, oxo, -C (O) Oalkyl, cycloalkyl, alkylene-O-alkyl, -C (O) haloalkyl, -C (O) -alkylene-O-alkyl, cyanoalkyl, alkylene-S (O)_xAlkyl, -alkylene-C (O) N (alkyl)₂Halogen, haloalkyl and alkoxy, wherein x is 0, 1 or 2.

As used herein, "heteroalkyl" refers to an alkyl group, as defined herein, wherein one or more of the constituent carbon atoms is replaced with nitrogen, oxygen, or sulfur. In some embodiments, heteroalkyl groups may also be substituted with 1,2, 3, or 4 substituents of alkyl groups as described herein. Examples of heteroalkyl substituents may include "alkoxy" as used herein, meaning alkyl-O- (e.g., methoxy and ethoxy). Heteroalkylene is a divalent heteroalkyl group.

As used herein, "alkenyl" refers to straight or branched chain alkyl groups having one or more carbon-carbon double bonds. Examples of the alkenyl group may include vinyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, and the like. The one or more carbon-carbon double bonds may be internal (e.g., double bond in 2-butene) or terminal (e.g., double bond in 1-butene). In various embodiments, the alkenyl group can have 2 to 40 carbon atoms (i.e., a C2-40 alkenyl group), for example, 2 to 20 carbon atoms (i.e., a C2-20 alkenyl group) or 2-20 carbon atoms (i.e., a C1-10 alkenyl group). In some embodiments, alkenyl groups may be substituted as described herein. An alkenyl group is typically not substituted with another alkenyl, alkyl, or alkynyl group.

As used herein, "fused ring" or "fused ring moiety" refers to a polycyclic ring system having at least two rings, wherein at least one ring is aromatic and such aromatic ring (carbocyclic or heterocyclic) has a bond in common with at least one other ring (which ring may be aromatic or non-aromatic and is carbocyclic or heterocyclic). These polycyclic systems may be highly p-conjugated and optionally substituted as described herein.

As used herein, "heteroatom" refers to an atom of any element other than carbon or hydrogen, and includes, for example, nitrogen, oxygen, silicon, sulfur, phosphorus, and selenium.

As used herein, "aryl" refers to an aromatic monocyclic hydrocarbon ring system or a polycyclic ring system in which two or more aromatic hydrocarbon rings are fused (i.e., have a common bond) together or at least one aromatic monocyclic hydrocarbon ring is fused with one or more cycloalkyl and/or cycloheteroalkyl rings. The aryl group can have 6 to 24 carbon atoms in its ring system (e.g., a C6-24 aryl group), which can include multiple fused rings. In some embodiments, the polycyclic aryl group can have 8 to 24 carbon atoms. Any suitable ring position of the aryl group may be as definedThe chemical structure is covalently linked. Examples of the aryl group having only an aromatic carbocyclic ring may include phenyl, 1-naphthyl (bicyclic), 2-naphthyl (bicyclic), anthracenyl (tricyclic), phenanthrenyl (tricyclic), condensed pentaphenyl (pentacyclic), and the like groups. Examples of polycyclic ring systems in which at least one aromatic carbocyclic ring is fused to one or more cycloalkyl and/or cycloheteroalkyl rings may include benzo derivatives of cyclopentane (i.e., indanyl, which is a 5, 6-bicyclic cycloalkyl/aryl ring system), benzo derivatives of cyclohexane (i.e., tetrahydronaphthyl, which is a 6, 6-bicyclic cycloalkyl/aryl ring system), benzo derivatives of imidazoline (i.e., benzimidazolinyl, which is a 5, 6-bicyclic cycloheteroalkyl/aryl ring system), and benzo derivatives of pyran (i.e., benzopyranyl, which is a 6, 6-bicyclic cycloheteroalkyl/aryl ring system). Other examples of aryl groups may include benzodioxanyl, benzodioxolyl, chromanyl, indolinyl, and the like. In some embodiments, aryl groups may be substituted as described herein. In some embodiments, an aryl group may have one or more halogen substituents, and may be referred to as a "haloaryl". Perhaloaryl, i.e. aryl in which all hydrogen atoms have been replaced by halogen atoms (e.g. -C)₆F₅) Included within the definition of "haloaryl". In certain embodiments, an aryl group is substituted with another aryl group and may be referred to as a biaryl group. Each aryl group in the biaryl group may be substituted as disclosed herein.

As used herein, a "diagnostic therapeutic agent" refers to an organic material having both diagnostic and therapeutic capabilities.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the presently described subject matter belongs.

Where a range of values is provided, for example, a concentration range, a percentage range, or a ratio range, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit and any other stated or intervening value in the stated range, is encompassed within the subject matter described. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and these embodiments are also encompassed within the described subject matter, subject to any specifically excluded limit in the stated range. Where a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the described subject matter.

Throughout this application, the description of the various embodiments uses the expression "comprising". However, those skilled in the art will understand that in some particular cases the expression "consisting essentially of or" consisting of "may be used instead to describe embodiments.

In order to better understand the teachings of the present invention and in no way limit the scope of the teachings thereof, all numbers expressing quantities, percentages or proportions, and other numerical values used in the specification and claims are to be understood as being modified in all instances by the term "about", unless otherwise indicated. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

Photosensitizers

The present subject matter relates to photosensitizers including compounds that exhibit an aggregation-induced ROS generation (aiorg) effect. Based on single crystal analysis, it is believed that the anion of this compound-pi⁺The interaction and the highly distorted conformation are responsible for the AIE effect (fig. 4A-4D). The inherent positive charge renders the molecule moderately water soluble and useful for biological applications. Furthermore, the compounds show a height of up to 100%¹O₂Quantum yield.

Photosensitizers can be used in photodynamic therapy (PDT) to selectively eliminate different targets without changing the structure of the compound. For example, time-dependent photodynamic therapy (PDT) can be achieved by controlling the incubation time of PS with different targets. The compounds can generate Reactive Oxygen Species (ROS) in photodynamic therapy (PDT) for selective bacterial eliminationAnd cancer cell ablation. For example, the compounds can be used as photosensitizers in PDT to generate singlet oxygen with high efficiency¹O₂)。

In one embodiment, the photosensitizer comprises a compound having the following backbone structure:

wherein R is₁Selected from the group consisting of H and alkyl;

wherein each R₂And R₃Independently selected from the group consisting of H, alkyl, unsaturated alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and diphenylamine; and is

X^-Selected from the group consisting of PF^6-、BF^4-、CH₃COO^-、CF₃COO^-、ClO^4-、F^-、Cl^-、Br^-、I^-、(F₃CSO₂)N^-And PO₄ ^3-Group (d) of (a).

With respect to the structural formulas of the compounds provided above, it is understood that R₂May be the same or different, and R₃May be the same or different. Furthermore, R₂And R₃May be the same as or different from each other.

In one embodiment, the compound is:

sufficiently small energy gap (Δ E) between excited singlet and excited triplet states_ST) Can be arranged in¹O₂Plays a major role in generating the required inter-system crossing (ISC) process. The compounds of the invention can exhibit extremely high levels of aggregation as compared with commercially available Photosensitizers (PS)¹O₂Quantum yield (fig. 6A to 6E and fig. 7A to 7D). For example, 4TPA-BQ exhibits in aggregate form¹O₂The quantum yield was about 98.7%.

Selectively killing cancer cells and eliminating pathogens

As set forth in detail herein, the compounds of the present invention can selectively target cancer cells rather than normal cells after incubation with cancer cells. The compounds of the present invention may also provide extremely high reactive oxygen (e.g., singlet oxygen) generation efficiency when exposed to white light illumination. Thus, the compounds of the present invention can be used as photosensitizers and provide selective cytotoxicity against cancer cells. In one embodiment, the compounds of the invention are useful for selective ablation of cancer cells. In image-guided PDT, one or more compounds of the present invention may be effective photosensitizers.

In one embodiment, a method of killing cancer cells can include incubating target cancer cells with one or more compounds of the present invention and, while the compounds are in contact with the target cancer cells, subjecting the target cancer cells to white light illumination to kill the target cancer cells. In one embodiment, the cancer cells are incubated with the compound for about 12 hours. In one embodiment, white light illumination of the target cancer cells may comprise using an illumination power of about 60mW cm^-2The lamp (2) was illuminated for about 30 minutes. In one embodiment, the target cancer cell is in a live animal.

For example, incubation of cancer cells with 4TPA-BQ for 12 hours by photodynamic therapy PDT specifically killed cancer cells with low toxicity to normal cells (fig. 11, fig. 16A to 16B, and fig. 17A to 17B).

In one embodiment, a method of killing a target pathogen may comprise incubating the target pathogen with one or more compounds of the invention and, while the compounds are in contact with the target pathogen, white light illumination of the target pathogen to kill the target pathogen. The target pathogen may be selected from at least one of gram-negative and gram-positive bacteria. The target pathogen may be incubated with the compound for about 15 minutes. In one embodiment, white light illumination of the target cancer cells may comprise using an illumination power of about 60mW cm^-2The lamp (2) was illuminated for about 30 minutes. The target pathogen may be in a living animal. The animal can be a human or a mammal.

For example, after incubation of 4TPA-BQ with bacteria for 15min, broad spectrum and potent antibacterial activity was obtained (fig. 10). After light exposure, 4TPA-BQ targeted and killed both gram-negative and gram-positive bacteria at low concentrations of 2 μ M, while normal cells were unaffected (fig. 12). Both in vitro and in vivo experiments showed that 4TPA-BQ has excellent bacterial elimination properties against ampicillin resistant E.coli infections (FIGS. 13A to 13F). It is believed that this time-dependent PDT is achieved by a synergistic effect of electrostatic attraction and hydrophobic interaction between 4TPA-BQ and the corresponding target.

The present teachings are illustrated by the following examples.

Examples

Materials and instruments

And (3) characterization: measurement with Bruker ARX 400NMR Spectroscopy using chloroform, MeOD or DMSO as solvents and tetramethylsilane (TMS;. delta. ═ 0) as internal standard¹H and¹³c NMR spectrum. High Resolution Mass Spectra (HRMS) were recorded using a Finnigan MAT TSQ7000 mass spectrometry system operating in MALDI-TOF mode. The absorption spectra were recorded using a Shimadzu UV-3600 spectrophotometer. By usingThe spectrophotometer records the Photoluminescence (PL) spectrum. Single crystal X-ray diffraction measurements were performed using a Bruker-Nonius Smart Apex CCD diffractometer with graphite monochromatic Mo Ka radiation. Photoluminescence quantum yield was determined using Hamamatsu absolute PL quantum yield spectrometer C11347 Quantaurus-QY. Transient PL at room temperature was measured using a Quantaurus-Tau fluorescence lifetime measurement system (C11367-03, Hamamatsu Photonics Co., Japan). Powder and film X-ray diffraction was performed using a Philips PW 1830X-ray diffractometer.

Example 1

Synthesis of Compound 1

Baking in proper directionBox dried 25mL round bottom flask to which Pd (PPh) was added₃)₂Cl₂(0.078mmol, 58mg), CuI (0.13mmol, 24.8mg) and 4-bromotriphenylamine (1.3mmol, 421.5mg), and the flask was purged with argon. Anhydrous toluene (8mL) and DBU (7.8mmol, 1.17mL) were added sequentially via syringe as the argon sparge. Ice-cold trimethylsilylacetylene (0.65mmol, 92uL) was then added via syringe followed immediately by distilled water (0.52mmol, 9.36 uL). The reaction flask was covered with aluminum foil and stirred at 80 ℃ for 20h at high speed. The reaction mixture was then extracted in DCM and distilled water. The organic layer was washed with 10% HCl and brine, and over MgSO₄And (5) drying. The crude product was purified by silica gel column chromatography with hexane/ethyl acetate (5:1, v/v) in 65% yield.¹H NMR(400MHz,CD₂Cl₂),δ(ppm):7.39–7.31(m,4H),7.34–7.23(m,8H),7.18–7.02(m,12H),7.03–6.94(m,4H)。¹³C NMR(100MHz,CD₂Cl₂),δ(ppm):147.08,146.58,131.59,129.00,128.73,124.33,123.14,122.90,121.78,121.59,115.71,88.06。HRMS(MALDI-TOF):m/z:[M+H]⁺C₃₈H₂₈N₂Calculated values: 512.2252, respectively; measured value: 512.2235.

example 2

Synthesis of 4TPA-BQ

To a 25mL pressure bottle were added 2-methylallylamine (0.15mmol, 14.03mg), 4' - ((1, 2-ethynyldiyl) bis [ N, N-diphenylaniline ]](0.45mmol, 229mg), copper acetate (0.75mmol, 149.7mg), [ Cp × RhCl₂]₂(0.0075mmol, 4.63mg), sodium tetrafluoroborate (0.225mmol, 24.7mg) and methanol. The resulting solution was stirred at 130 ℃ overnight and over anhydrous MgSO₄And (5) drying. The crude product was purified by silica gel column chromatography with DCM: MeOH (25:1, v: v) in 81% yield.¹H NMR(400MHz,CD₂Cl₂),δ(ppm):8.60(s,1H),7.97(s,1H),7.90(d,J＝9.6Hz,1H),7.42–7.27(m,21H),7.21–7.03(m,24H),6.98–6.96(m,5H),6.89–6.84(m,4H),2.50(s,3H)。¹³C NMR(100MHz,CD₂Cl₂),δ(ppm):152.03,150.03,149.72,148.12,147.91,147.56,147.30,145.33,143.39,142.17,139.33,139.06,137.01,136.34,133.37,133.08,132.35,131.79,131.28,131.19,130.50,130.34,130.23,130.10,129.95,128.37,127.24,126.88,126.15,125.82,125.13,124.98,124.60,123.96,123.57,123.12,122.10,120.31,118.24,112.78,18.94.HRMS(MALDI-TOF):m/z:[M-PF6]⁺C₈₀H₆₀N₅ ⁺Calculated values: 1090.4843, respectively; measured value: 1090.4847.

an exemplary reaction scheme for preparing the 4TPA-BQ compound can include a one-step synthetic process, as follows:

example 3

Photophysical Properties of 4TPA-BQ

4TPA-BQ was well characterized by NMR, high resolution mass spectrometry and single crystal X-ray diffraction analysis (Table 1). The data obtained have good agreement with the proposed structure (fig. 1 to 3, fig. 5).

TABLE 1.4 photophysical properties of TPA-BQ

Will be provided with^aτ is defined by the formula τ ═ Σ a_i(τ_i)²/ΣA_iτ_iThe calculated average fluorescence lifetime is calculated as the average fluorescence lifetime,

wherein A is_iPre-exponential factor tau for life_i。^bk_r＝Φ/τ。^ck_nr＝1/τ-k_r。

The subject matter of the invention is described above, it being obvious that it may be modified or varied in many ways. Such modifications and variations are not to be regarded as a departure from the spirit and scope of the inventive subject matter, and all such modifications and variations are intended to be included within the scope of the following claims.