Chiral cuprous sulfide nano-catalytic particles, and preparation method and application thereof

Abstract

The application discloses chiral cuprous sulfide nano-catalytic particles, a preparation method and application thereof. Copper salt, D-penicillamine or L-penicillamine and sodium citrate are mixed in an aqueous phase, sodium borohydride is added, and reduction self-assembly under coordination induction is carried out under neutral to weak alkaline conditions and in a protective atmosphere to obtain chiral cuprous sulfide nano-particles in D-configuration or L-configuration; further surface modification by polyethylene glycol-phenylphosphine can obtain D-Cu2S-TPP or L-Cu2S-TPP nano-catalytic particles with mitochondria targeting function. The nano-catalytic particles can be used for catalytic conversion of hydrogen peroxide and in-situ generation of hydroxyl radicals, and can also be used for preparing a drug composition for inducing tumor cell ferroptosis, which plays a role by enhancing intracellular lipid peroxidation. The application discloses the role of chiral configuration in regulating nano-catalytic Fenton reaction, and provides a new technical scheme for design and application of configuration-dependent nano-catalysts.

Description

Technical Field

[0001] This invention belongs to the field of nanocatalytic materials and biomedical materials technology, specifically relating to a chiral engineered cuprous sulfide nanocatalytic particle constructed with penicillamine, a method for preparing the nanocatalytic particle, and its application in the catalytic activation of hydrogen peroxide and the preparation of compositions that induce ferroptosis in tumor cells. Background Technology

[0002] Chirality, as a pervasive structural feature embedded in living systems, underpins stereoselective molecular recognition, catalytic specificity, membrane-associated signaling, immune responses, and cell fate regulation. This inherent stereochemical bias stems from the homochiral organization of biological building blocks, such as L-amino acid-based proteins, D-carbazide-containing nucleic acids, and chiral phospholipid membranes, enabling biological interfaces to distinguish mirror-image structures with high selectivity. In medicinal chemistry, the two enantiomers of chiral compounds can exhibit significantly different biological activities, with one producing the desired therapeutic effect while its mirror-image counterpart remains inactive or even toxic. Inspired by the central role of chirality in biological systems, nanoparticles with tunable size and surface chemistry offer unique opportunities to amplify chiral-mediated biological responses. Against this backdrop, chiral nanoparticles designed via ligand-mediated synthesis or chiral transfer strategies have recently attracted increasing interest because their stereochemical configurations can elicit diverse biological outcomes.

[0003] Hydrogen peroxide (H₂O₂) is an important reactant in various oxidation reactions and biological oxidative stress processes. Cuprous sulfide containing Cu(I) active sites can promote the cleavage of the OO bonds in hydrogen peroxide and generate highly reactive hydroxyl radicals through a Fenton-like reaction. Compared with free copper ions, nanoscale cuprous sulfide is easier to construct local catalytic interfaces, and its dispersibility and biocompatibility can be improved through surface modification.

[0004] Existing technologies for improving Fenton-like catalysis efficiency mainly focus on elemental doping, valence state modulation, defect construction, particle size optimization, and cascade reaction design. These approaches typically treat the catalytic interface as an achiral environment, paying less attention to the influence of the stereoconfiguration of surface ligands on reactant capture, orientation, and effective collisions. Therefore, under conditions of essentially the same composition and particle size, whether it is possible to regulate hydrogen peroxide activation efficiency solely through the chiral interface remains a question lacking a clear and feasible material approach.

[0005] Both D-penicillamine and L-penicillamine contain thiol groups that can coordinate with copper and have mirror-image stereoconfigurations. Using them in the interface construction of cuprous sulfide nanoparticles allows for the acquisition of D- and L-configurations while maintaining similar bulk chemical compositions, providing a basis for comparing and utilizing configuration-dependent catalytic performance. It should be noted that hydrogen peroxide itself is an achiral molecule; the "stereochemical regulation" mentioned in this invention refers to the configuration-dependent influence of chiral interfaces on the adsorption, orientation, and activation processes of hydrogen peroxide, rather than the separation of hydrogen peroxide enantiomers.

[0006] Therefore, it is necessary to provide a cuprous sulfide nanocatalytic particle with a well-defined composition, reproducible preparation conditions, and the ability to regulate the catalytic activation of hydrogen peroxide through chiral configuration, and to further establish the correspondence between its material structure, catalytic performance and biological effects. Summary of the Invention

[0007] This invention aims to address the shortcomings of existing cuprous sulfide-based Fenton nanocatalysts, which primarily rely on chemical composition or defect regulation and lack the utilization of chiral interfaces to enhance hydrogen peroxide capture and catalytic activation efficiency. It provides a nanocatalytic material capable of stably preparing a D / L mirror-image configuration, where the L configuration exhibits higher hydroxyl radical generation capacity. This invention utilizes copper ions (Cu... + Leveraging the catalytic advantages of copper nanoparticles, we rationally constructed chiral copper nanoparticles to serve as stereochemically tunable Fenton-like nanocatalysts for evaluating chiral-regulated •OH generation and ferroptosis-mediated antitumor therapy.

[0008] The technical solution adopted in this invention is as follows:

[0009] A chiral cuprous sulfide nanocatalytic particle, wherein the core of the nanocatalytic particle is cuprous sulfide Cu2S; the surface of Cu2S is bound to chiral ligands D-penicillamine or L-penicillamine (D / L-PEN) through thiol coordination bonds to obtain D / L-Cu2S nanoparticles; further functionalized with PEG-TPP to obtain mitochondrial-targeted D-Cu2S-TPP or L-Cu2S-TPP nanoparticles.

[0010] The nanoparticles are spherical or near-spherical in shape, with an average diameter of 75±5 nm.

[0011] The chiral ligands D-penicillamine or L-penicillamine directly participate in the construction of nanoparticles during the reductive self-assembly process. The chiral coordination layer is formed on the surface of cuprous sulfide nanoparticles by the coordination of D-penicillamine or L-penicillamine with copper through the coordination of thiol groups. The nanocatalytic particles have a circular dichroism signal corresponding to the D-penicillamine or L-penicillamine used.

[0012] The nanocatalytic particles possess chiral stereochemistry-dependent Fenton-like catalytic activity. Compared to D-Cu2S-TPP, L-Cu2S-TPP has a stronger H2O2 activation ability, which can stereoselectively amplify the generation of hydroxyl radicals (•OH) and achieve enantioselective induction of ferroptosis in tumor cells.

[0013] The preparation method of the chiral cuprous sulfide nanocatalytic particles includes the following steps:

[0014] (1) Cu + A copper source is mixed with chiral ligands D-PEN or L-PEN, utilizing the thiol groups of D / L-PEN and Cu... + Thiol-mediated coordination between them leads to reductive self-assembly under the guidance of chiral ligands, generating D / L-Cu2S nanoparticles.

[0015] (2) The D / L-Cu2S nanoparticles obtained in step (1) are functionalized by grafting with PEG-TPP and purified to obtain mitochondrial-targeted D-Cu2S-TPP or L-Cu2S-TPP nanocatalytic particles.

[0016] The copper source is a monovalent copper salt, selected from at least one of cuprous chloride and cuprous acetate; the reduction self-assembly is carried out in an aqueous system.

[0017] The number-average molecular weight of the polyethylene glycol is 1500–2500 Da.

[0018] More specifically, in step (1), copper salt, D-penicillamine or L-penicillamine and sodium citrate are dispersed in water and mixed for 20 to 40 min; sodium borohydride is added and the pH of the reaction system is adjusted to 7.0 to 8.0; the reaction is carried out under a protective atmosphere for 8 to 18 h, and after separation and purification, D-configuration or L-configuration chiral cuprous sulfide nanocatalytic particles are obtained.

[0019] In step (2), mitochondrial targeting function modification: chiral Cu2S nanoparticles and TPP-PEG-SH targeting molecules are uniformly dispersed in the aqueous phase, stirred at room temperature for 36-60 h, and purified by dialysis to obtain L-Cu2S-TPP or D-Cu2S-TPP.

[0020] The copper salt mentioned in step (1) is copper chloride; the molar ratio of copper chloride, D-penicillamine or L-penicillamine, sodium citrate and sodium borohydride is 1:(0.8-1.2):(0.8-1.2):(1.0-1.5).

[0021] The protective atmosphere in step (1) is nitrogen; the separation and purification includes mixing the reaction solution with acetone at a volume ratio of 1:(5-9) and centrifuging at 10000-14000 rpm for 10-20 min.

[0022] In step (2), the mass ratio of D / L-Cu2S nanoparticles to PEG-TPP is (4-6):1.

[0023] Functionalization step: The obtained chiral cuprous sulfide nanocatalyst particles were dispersed in water with mercapto-polyethylene glycol-triphenylphosphine at a mass ratio of (4-6):1, and reacted at room temperature for 36-60 h. After purification by dialysis and / or ultrafiltration, D-Cu2S-TPP or L-Cu2S-TPP nanocatalyst particles with mitochondrial targeting function were obtained.

[0024] This invention further provides the application of these nanocatalytic particles in the catalytic activation of hydrogen peroxide. The L-configuration particles can form a more favorable hydrogen peroxide interfacial environment near the catalytically active site, improving the local enrichment, stable orientation, and effective collision probability of hydrogen peroxide, thereby increasing the transient generation of surface-confined hydroxyl radicals.

[0025] Application of the chiral cuprous sulfide nanocatalyst particles in the catalytic activation of hydrogen peroxide to generate hydroxyl radicals.

[0026] In the above applications, the catalytic reaction is carried out in an aqueous system with pH 4.5–7.5 and temperature 25–40 °C; the concentration of the nanocatalytic particles is 50–200 μg / mL, and the concentration of hydrogen peroxide is 1–20 mM.

[0027] The chiral cuprous sulfide nanocatalyst particles are L-Cu2S-TPP nanocatalyst particles, and their chiral interface is used to improve the capture and activation efficiency of hydrogen peroxide near the catalytic active site.

[0028] In biological applications, the aforementioned nanocatalytic particles, especially L-Cu2S-TPP, can be used to prepare compositions that induce ferroptosis in tumor cells. Their mechanism of action includes enhancing intracellular oxidative stress, promoting lipid peroxidation, reducing glutathione-related antioxidant capacity, and inducing mitochondrial dysfunction.

[0029] An antitumor drug composition comprising the chiral cuprous sulfide nanocatalytic particles and pharmaceutically acceptable excipients.

[0030] A pharmaceutical composition for inducing ferroptosis in tumor cells comprises the aforementioned chiral cuprous sulfide nanocatalytic particles and a pharmaceutically acceptable carrier; wherein the chiral cuprous sulfide nanocatalytic particles are preferably L-Cu2S-TPP nanocatalytic particles.

[0031] The dosage form of the pharmaceutical composition is any one of injection, lyophilized powder for injection, or nano-dispersion.

[0032] In the above pharmaceutical composition, the chiral cuprous sulfide nanocatalytic particles are preferably L-Cu2S-TPP.

[0033] Compared to D-configuration nanoparticles, L-configuration nanoparticles exhibit superior stereoselectivity in catalytic H2O2-mediated hydroxyl radical (•OH) generation. L-configuration nanoparticles demonstrate enhanced Fenton-like catalytic activity for H2O2 activation, inducing higher levels of surface-confined •OH. This chirality-dependent catalytic amplification effect induces severe mitochondrial dysfunction, promotes lipid peroxide accumulation, and activates ferroptosis by regulating key ferroptosis-related pathways. Therefore, L-Cu2S-TPP exhibits significantly enhanced ferroptosis cytotoxicity in vitro and superior tumor growth inhibition in vivo.

[0034] A stereochemically regulated nanocatalytic strategy for the generation of endogenous hydroxyl radicals and targeted induction of tumor ferroptosis is proposed. L-Cu2S-TPP nanomaterials are constructed using L-penicillamine coordination. The unique chiral interface structure of L-Cu2S-TPP optimizes the H2O2 activation kinetics, amplifies the generation of surface-confined •OH, and activates tumor cell ferroptosis in a stereoselective manner to achieve targeted anti-tumor therapy.

[0035] Density functional theory (DFT) and molecular dynamics (MD) simulations revealed that L-configuration nanoparticles can efficiently catalyze the generation of higher levels of surface-confined •OH from endogenous hydrogen peroxide (H₂O₂). This enhanced catalytic activity is primarily attributed to higher molecular collision efficiency, more stable capture and orientation of H₂O₂ molecules, and an optimized reaction microenvironment associated with the L-configuration, thereby achieving chiral-dependent regulation of the Fenton-like reaction. Furthermore, in vitro and in vivo studies demonstrated that L-Cu₂S-TPP enhances •OH generation, leading to glutathione peroxidase 4 (GPX-4) dysregulation and lethal accumulation of lipid peroxides (LPO), thereby inducing ferroptosis and tumor cell death. Therefore, this study confirms that the designed chiral cuprous sulfide nanocatalytic particles can utilize stereochemistry to promote the generation of Fenton-like •OH, providing a mechanistic basis and novel strategy for chiral-mediated H₂O₂ activation regulation to induce ferroptosis.

[0036] Beneficial effects:

[0037] (1) The present invention uses D / L-penicillamine, which are mirror images of each other, as the interface building unit, and can obtain cuprous sulfide nanocatalytic particles with mirror circular dichroism signals without introducing different metal elements.

[0038] (2) Under the same reaction conditions, the L-configuration nanocatalytic particles have a higher catalytic activation ability for hydrogen peroxide than the D-configuration particles; the results of methylene blue degradation and electron spin resonance both indicate that the L-configuration can generate more hydroxyl radicals.

[0039] (3) The preparation process of the present invention is carried out under aqueous phase, neutral to weakly alkaline and room temperature conditions, and the raw materials and steps are clearly defined, which makes it easy to repeat the process.

[0040] (4) By modifying with polyethylene glycol-triphenylphosphine, the particles can be given good water dispersibility and mitochondrial localization ability, so that the configuration-dependent catalytic difference can be further transformed into lipid peroxidation and ferroptosis-induced difference. Attached Figure Description

[0041] Figure 1 TEM images of L-Cu2S-TPP (A) and D-Cu2S-TPP (B).

[0042] Figure 2 HR-TEM images and corresponding lattice structure analysis of L-Cu2S-TPP.

[0043] Figure 3 HAADF and EDS elemental mapping images of N, O, S and Cu in L-Cu2S-TPP.

[0044] Figure 4 CD spectra of L-Cu2S-TPP and D-Cu2S-TPP.

[0045] Figure 5 DLS curves of L-Cu2S-TPP and D-Cu2S-TPP.

[0046] Figure 6 XRD patterns of D-Cu2S and L-Cu2S.

[0047] Figure 7 Fourier transform infrared (FT-IR) spectrum.

[0048] Figure 8 Ultraviolet-Vis (UV-vis) spectrum

[0049] Figure 9 (A) Time-dependent degradation curves of MB treated with D-Cu2S-TPP and L-Cu2S-TPP. (B) ESR spectra of DMPO-OH adducts generated by D-Cu2S-TPP and L-Cu2S-TPP, and (C) corresponding quantitative analysis of •OH formation. (D) DFT calculations of the reaction mechanism of H2O2 activation and surface-limited •OH formation on Cu2S catalytic surface. Optimized geometric configurations of intermediates (INT1-INT3) and transition states (TS). HAT represents hydrogen atom transfer. (E) Corresponding Gibbs free energy distribution of the reaction pathway.

[0050] Figure 10Molecular dynamics simulations revealed the chirality-dependent interactions between H₂O₂ and L- or D-Cu₂S-TPP. Representative molecular dynamics simulations show the most stable interaction structures between (A) L-Cu₂S-TPP and H₂O₂, and (B) D-Cu₂S-TPP and H₂O₂. (C) Electrostatic interactions (Ele) between H₂O₂ and L- or D-Cu₂S-TPP. (D) Van der Waals interactions (vdW) between H₂O₂ and L- or D-Cu₂S-TPP. (E) Mean square shift (MSD) curves of the interaction between H₂O₂ and L- or D-Cu₂S-TPP. (F) Time-dependent changes in the number of hydrogen bonds formed between H₂O₂ and L- or D-Cu₂S-TPP. (G) Distribution of hydrogen bond distances between H₂O₂ and L- or D-Cu₂S-TPP. (H) Thermodynamic parameters (ΔH, ΔS and ΔG) of the interaction between H2O2 and L- or D-Cu2S-TPP based on molecular dynamics simulations.

[0051] Figure 11 (A) Colocalization of L-Cu2S-TPP and L-Cu2S with mitochondria in KYSE-150 cells stained with Mito Tracker Green, measured by CLSM. Red fluorescence indicates NPs. Scale bar: 25 μm. (B) Quantitative analysis of mitochondrial colocalization. Pearson correlation coefficient between NP fluorescence and Mito Tracker Green fluorescence in KYSE-150 cells.

[0052] Figure 12 CLSM images and corresponding surface maps of ROS levels in KYSE-150 cells after different treatments. Scale bar: 50 μm.

[0053] Figure 13 Transmission electron microscopy images of mitochondrial morphology in KYSE-150 cells after different treatments. Scale bar: 2 μm.

[0054] Figure 14 The expression levels of ACSL4, GPX-4 and FSP1 were analyzed by Western blotting.

[0055] Figure 15 :BODIPY TM 665 / 676 The distribution of LPO in KYSE-150 cells treated with L-Cu2S-TPP and L-Cu2S-TePP+Fer-1 was detected by staining. Scale bar: 50 μm. Detailed Implementation

[0056] The following examples are used to further illustrate the present invention, but should not be construed as limiting the scope of protection of the present invention. Operations not specifically specified can be performed under conventional conditions in the art. In the examples, D / L indicates that the corresponding products are prepared using D-configuration or L-configuration raw materials respectively, rather than a mixture of the two.

[0057] D-Penicillamine (D-PEN), L-Penicillamine (L-PEN), CuCl2·2H2O, sodium citrate, NaBH4, H2O2, Cy5.5, N-hydroxysuccinimide (NHS), sodium hydroxide (NaOH), N-ethylcarbodiimide hydrochloride (EDCl), methylene blue (MB), and iron stabilizer-1 (Fer-1) were all purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. Triphenylphosphine-PEG2000-mercapto (TPP-PEG2000-SH) was purchased from Shanghai Pengsheng Biotechnology Co., Ltd.

[0058] Example 1: Preparation of D-Cu2S and L-Cu2S nanoparticles

[0059] All glassware was washed with freshly prepared aqua regia and rinsed thoroughly with deionized water. In a three-necked flask, 0.6 mL of a 1 mol / L CuCl2·2H2O aqueous solution was added to 8 mL of deionized water; then 1.2 mL of a 0.5 mol / L D-penicillamine or L-penicillamine aqueous solution and 0.4 mL of a 1.5 mol / L sodium citrate aqueous solution were added, and the mixture was stirred vigorously at room temperature for 30 min.

[0060] Add 0.5 mL of a 1.5 mol / L NaBH4 aqueous solution to the above system, adjust the pH to 7.5 with 0.1 mol / L NaOH, and magnetically stir at room temperature for 12–16 h under nitrogen protection to obtain a deep purple dispersion. Mix the obtained dispersion with acetone at a volume ratio of 1:7, centrifuge at 12000 rpm for 15 min, collect the precipitate, and obtain D-Cu2S and L-Cu2S nanoparticles, respectively.

[0061] Based on the amount of feed, the molar ratio of CuCl2·2H2O, penicillamine, sodium citrate, and NaBH4 is 1:1:1:1.25. When using D-penicillamine and L-penicillamine, the amounts of feed and reaction conditions remain consistent, except for the ligand configuration.

[0062] Example 2 Preparation of D-Cu2S-TPP and L-Cu2S-TPP

[0063] Weigh out 50 mg of D-Cu2S or L-Cu2S prepared in Example 1, and mix with 10 mg of TPP-PEG. 2000-SH were co-dispersed in 8 mL of deionized water and magnetically stirred at room temperature for 48 h. Dialysis was performed using a dialysis bag with a molecular weight cutoff of 2000 Da, and unbound molecules were removed by ultrafiltration to obtain D-Cu2S-TPP or L-Cu2S-TPP aqueous dispersions, which were stored at 4 °C.

[0064] Transmission electron microscopy results showed that both types of particles were relatively uniformly spherical or near-spherical, with an average particle size of approximately 75 ± 5 nm. Dynamic light scattering measurements revealed that the hydrated particle sizes of D-Cu₂S-TPP and L-Cu₂S-TPP were approximately 105 nm and 114 nm, respectively. The similar particle size and zeta potential of both types of particles suggest that the subsequent differences in catalytic performance can be primarily attributed to the chiral interface rather than significant size or charge differences.

[0065] Example 3: Structure and Chirality Characterization

[0066] The particles obtained in Example 2 were characterized by transmission electron microscopy, high-resolution transmission electron microscopy, energy dispersive X-ray spectroscopy, circular dichroism spectroscopy, X-ray photoelectron spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and ultraviolet-visible absorption spectroscopy.

[0067] High-resolution images of L-Cu₂S-TPP show a crystal plane spacing of approximately 343.4 pm, and elemental mapping reveals the distribution of N, O, S, and Cu within the particles. D-Cu₂S-TPP and L-Cu₂S-TPP exhibit circular dichroism signals with opposite directions and corresponding shapes in the 250–650 nm range, demonstrating that both possess mirror-chiral optical responses.

[0068] D / L penicillamine at 2500–2600 cm⁻¹ -1 The thiol stretching vibration peak at the thiol group disappears after the formation of D / L-Cu2S, indicating that the thiol group participates in Cu-S coordination or bonding; TPP-PEG 2000 The value 2889 cm appears after the -SH modifier. -1 and 2858 cm -1 The nearby methylene stretching vibration peak confirms the introduction of the functional modification layer.

[0069] Example 4: Catalytic activation performance of hydrogen peroxide

[0070] Aqueous dispersions of D-Cu₂S-TPP and L-Cu₂S-TPP (100 μg / mL) were prepared and stirred at 37 °C under nitrogen protection. Methylene blue and H₂O₂ were added to bring the final H₂O₂ concentration to 10 mmol / L. The absorbance at 664 nm was measured at 0, 3, 6, 9, 12, and 15 min, and the residual methylene blue concentration was used to evaluate hydroxyl radical-mediated oxidative degradation.

[0071] The results showed that, within the same reaction time, the residual methylene blue content in the L-Cu2S-TPP group was lower than that in the D-Cu2S-TPP group; in the representative endpoint results, the residual methylene blue content in the D-configuration group and the L-configuration group was approximately 39% and 20%, respectively. This result indicates that the L-configuration particles have a higher catalytic activation capacity for hydrogen peroxide.

[0072] Furthermore, 100 μg / mL of D-Cu₂S-TPP or L-Cu₂S-TPP was reacted with 10 mmol / L H₂O₂ and 50 mmol / L 5,5-dimethyl-1-pyrroline-N-oxide, respectively, at pH 5.0 for 10 min, and electron spin resonance detection was performed. Both groups showed the characteristic 1:2:2:1 quartet of the DMPO-OH adduct, with the L-Cu₂S-TPP group exhibiting a higher signal intensity than the D-Cu₂S-TPP group. The results of methylene blue degradation and electron spin resonance detection, two independent methods, corroborated each other.

[0073] Example 5 Catalytic Mechanism Analysis

[0074] Density functional theory calculations were performed using the Cu(I)-penicillamine coordination site as a model. The results show that H₂O₂ first adsorbs at the Cu(I) site to form an intermediate, followed by electron transfer and OO bond cleavage, forming Cu(II)-OH and hydroxyl radicals near the active site; the calculated energy barrier for the OO bond cleavage step is approximately 28.8 kcal / mol. These hydroxyl radicals have short lifetimes and high reactivity, mainly exhibiting transient surface-confined species generated near the active site.

[0075] Molecular dynamics simulations further revealed that, compared to the D-configuration system, the L-configuration system exhibits a more negative interaction energy between the H₂O₂ and nanoparticle interfaces, resulting in a greater number of hydrogen bonds with shorter and more concentrated bond lengths; its enthalpy change and Gibbs free energy changes are also more favorable. These results support the view that the L-configuration interface can enhance the interfacial trapping, orientation stability, and effective collision frequency of H₂O₂.

[0076] It should be noted that density functional theory is used to explain the basic reaction pathway of Cu(I) site activation of H2O2, and molecular dynamics is used to compare the influence of different chiral interfaces on the interfacial behavior of H2O2; together with the experimental results of Example 4, the two constitute the evidence chain of material configuration-interfacial interaction-catalytic performance.

[0077] Example 6: Verification of the role of mitochondrial localization and ferroptosis

[0078] Cy5.5-labeled L-Cu2S or L-Cu2S-TPP were incubated with KYSE-150 cells for 4 h, and mitochondria were labeled with Mito-Tracker Green. Confocal microscopy results showed that the Pearson correlation coefficient between L-Cu2S-TPP and mitochondrial signaling was approximately 0.88, higher than the 0.41 of unmodified L-Cu2S, indicating that TPP modification improved the mitochondrial localization ability of particles. D-Cu2S-TPP showed similar mitochondrial localization to L-Cu2S-TPP, ruling out significant targeting differences as the main reason for the difference in L / D catalytic effects.

[0079] At the same dosage, L-Cu2S-TPP induced higher levels of intracellular ROS signaling, decreased mitochondrial membrane potential, and lipid peroxidation than D-Cu2S-TPP. The addition of 5 μmol / L ferrostatin-1 alleviated the L-Cu2S-TPP-induced decrease in cell viability, mitochondrial damage, and lipid peroxidation.

[0080] Protein analysis showed that ACSL4 expression was upregulated after L-Cu2S-TPP treatment, while GPX-4 and FSP1 expression were downregulated. Combined with the ferrostatin-1 rescue results, this indicates that the composition can promote ferroptosis through configuration-dependent hydrogen peroxide activation and lipid peroxidation. The above biological results are used to demonstrate the usability of the catalytic material, and do not use changes in the expression of a single protein as the sole criterion for determining ferroptosis.

[0081] D-Cu2S-TPP and L-Cu2S-TPP are similar in morphology, particle size, and mitochondrial localization, but D-Cu2S-TPP exhibits lower methylene blue degradation and DMPO-OH signal under the conditions described in Example 4, demonstrating that the L / D catalytic difference cannot be explained solely by macroscopic particle size or TPP targeting differences. Industrial Applicability

[0082] The raw materials used in this invention are readily available, and the reaction conditions are mild, enabling the production of both D-configuration and L-configuration cuprous sulfide nanocatalytic particles using the same process. The resulting materials can be used as configuration-dependent hydrogen peroxide catalytic activators for the research and preparation of aqueous oxidation reactions, in-situ hydroxyl radical generation systems, and ferroptosis-related drug compositions, demonstrating reproducibility and application value.

Claims

1. A chiral cuprous sulfide nanocatalytic particle, characterized in that, The core of the nanocatalytic particle is a cuprous sulfide nanoparticle and a chiral coordination layer bound to the surface of the cuprous sulfide nanoparticle; the chiral coordination layer is formed by the coordination of D-penicillamine or L-penicillamine with copper through the coordination of thiol groups, and the nanocatalytic particle has a circular dichroism signal corresponding to the D-penicillamine or L-penicillamine used.

2. The chiral cuprous sulfide nanocatalyst particles according to claim 1, characterized in that, The nanoparticles are spherical or near-spherical in shape, and the average particle size measured by transmission electron microscopy is 75±5 nm.

3. The method for preparing the chiral cuprous sulfide nanocatalytic particles according to claim 1, characterized in that, Includes the following steps: (1) Mix the copper source with the chiral ligand D-PEN or L-PEN, and utilize the D / L-PEN thiol group to react with Cu + Thiol-mediated coordination between them leads to reductive self-assembly under the guidance of chiral ligands, generating D / L-Cu2S nanoparticles. (2) The D / L-Cu2S nanoparticles obtained in step (1) are functionalized by grafting with PEG-TPP and purified to obtain mitochondrial-targeted D-Cu2S-TPP or L-Cu2S-TPP nanocatalytic particles.

4. The preparation method according to claim 3, characterized in that, The copper source is a monovalent copper salt, selected from at least one of cuprous chloride and cuprous acetate; the reduction self-assembly is carried out in an aqueous system.

5. The preparation method according to claim 3, characterized in that, In step (1), the molar ratio of copper chloride, D-penicillamine or L-penicillamine is 1:(0.8 to 1.2).

6. The preparation method according to claim 3, characterized in that, In step (2), the mass ratio of D / L-Cu2S nanoparticles to PEG-TPP is (4-6):

1.

7. The application of the chiral cuprous sulfide nanocatalyst particles according to claim 1 or 2 in the catalytic activation of hydrogen peroxide to generate hydroxyl radicals.

8. The application according to claim 7, characterized in that, The catalytic reaction was carried out in an aqueous system with pH 4.5–7.5 and temperature 25–40 °C; the concentration of the nanocatalytic particles was 50–200 μg / mL, and the concentration of hydrogen peroxide was 1–20 mM.

9. The use of the chiral cuprous sulfide nanocatalytic particles according to claim 1 or 2 in the preparation of a pharmaceutical composition that induces ferroptosis in tumor cells.

10. A pharmaceutical composition for inducing ferroptosis in tumor cells, characterized in that, It comprises the chiral cuprous sulfide nanocatalyst particles as described in claim 1 or 2 and a pharmaceutically acceptable support; the chiral cuprous sulfide nanocatalyst particles are preferably L-Cu2S-TPP nanocatalyst particles.

Images