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CN-122012493-A - Porphyrin-G quadruplex complex and application thereof in diagnosis and treatment of tumors

CN122012493ACN 122012493 ACN122012493 ACN 122012493ACN-122012493-A

Abstract

The invention relates to a porphyrin-G quadruplex complex and application thereof in diagnosis and treatment of tumors. Specifically, the invention provides a nucleic acid nanocluster, which comprises a nucleic acid aptamer AS 1411G-quadruplex sequence and a human telomerase-derived G-quadruplex sequence, can be used for loading porphyrin molecules such AS photosensitizer Ce6 and Her with peroxidase-like effect, and can be used for obtaining a porphyrin-nucleic acid nanocluster compound, so AS to play a role in diagnosing and/or treating tumors. The nucleic acid nanocluster provided by the invention has better targeting property, stability and higher drug loading capacity. The nucleic acid nanocluster loaded with Ce6 and Hem can realize the synergistic effect of multiple functions and has the enhanced function of inhibiting tumors.

Inventors

  • GUO GANG
  • CHEN BO
  • HAN BO

Assignees

  • 四川大学

Dates

Publication Date
20260512
Application Date
20241108

Claims (10)

  1. 1. A nucleic acid nanocluster is characterized in that the nucleic acid nanocluster is formed by agglomerating single-stranded nucleic acid, the single-stranded nucleic acid is formed by connecting a plurality of G-quadruplex (G4) structures in series, and the single-stranded nucleic acid comprises (1) a nucleic acid aptamer AS 1411G-quadruplex (AS 1411G 4) structural unit and (2) a human telomerase-derived G-quadruplex (Telo G) structural unit.
  2. 2. The nucleic acid nanocluster of claim 1, wherein the nucleic acid nanocluster is formed by rolling circle amplification from a template strand comprising (1) a complement of AS 1411G 4 and (2) a complement of Telo G4.
  3. 3. The nucleic acid nanocluster of claim 2 wherein the template strand has a structure according to formula I: Z1-G1-L-G2-Z2 (formula I) In the formula, Z1 and Z2 are primer segments; g1 is the complement of AS 1411G 4; L is none or a linker; g2 is the complement of Telo G4.
  4. 4. A porphyrin-nucleic acid nanocluster complex comprising the nucleic acid nanocluster of claim 1 and one or more porphyrin molecules, wherein the porphyrin molecules are incorporated into the G-quadruplex structure of the nucleic acid nanocluster.
  5. 5. The porphyrin-nucleic acid nanocluster complex of claim 4 wherein the porphyrin molecule is selected from the group consisting of chlorin e6 (Ce 6), methemoglobin (Hem), and combinations thereof.
  6. 6. A pharmaceutical composition comprising (1) the porphyrin-nucleic acid nanocluster complex of claim 4, and (2) a pharmaceutically acceptable carrier.
  7. 7. A complexing agent comprising AS 1411G 4 and Telo G molecules, wherein the AS 1411G 4 and Telo G molecules are present AS a single molecule or wherein the AS 1411G 4 and Telo G molecules are linked by a linker.
  8. 8. The use of the porphyrin-nucleic acid nanocluster complex as described in claim 4, for preparing a pharmaceutical composition for diagnosing and/or treating tumors.
  9. 9. A method of preparing the nucleic acid nanocluster of claim 1, comprising the steps of: (a) Providing a template strand comprising the complement of the AS 1411G 4 and Telo G4 sequences; (b) The template strand rolling circle amplification produces the nucleic acid nanoclusters under the reaction conditions of rolling circle amplification.
  10. 10. A method of preparing the porphyrin-nucleic acid nanocluster complex of claim 4, comprising the steps of: (i) Preparing the nucleic acid nanocluster of claim 1; (ii) Mixing a porphyrin molecule with the nucleic acid nanocluster to form the porphyrin-nucleic acid nanocluster complex.

Description

Porphyrin-G quadruplex complex and application thereof in diagnosis and treatment of tumors Technical Field The invention relates to the field of biological medicine, in particular to a porphyrin-G quadruplex complex and application thereof in diagnosis and treatment of tumors. Background The statistics show that the global cancer onset cases in 2020 are about 1930 ten thousand and the death cases are about 1000 ten thousand. Among them, three categories with the greatest number of diagnosis are female breast cancer (226 ten thousand), lung cancer (221 ten thousand) and prostate cancer (141 ten thousand). Although the incidence and mortality rate of cancer is reduced compared to the peak in 1991, it remains a global public medical problem. Breast cancer is a high-grade malignant tumor in females, has multiple types, and Triple Negative Breast Cancer (TNBC) is representative of high metastasis and poor prognosis, accounting for 11-23% of the total incidence rate of breast cancer. It shows negative Estrogen Receptor (ER), progestogen Receptor (PR) and human epidermal growth factor receptor 2 (HER 2), and the common treatment modes such as hormone treatment have poor effect due to lack of corresponding targets. And because TNBC has high transferability, the common clinical operation excision combined with radiotherapy and chemotherapy mode can not effectively solve the problems of transfer and recurrence, and can not show good curative effect. Photodynamic therapy (Photodynamic Therapy, PDT for short) is a novel therapy for treating diseases using photosensitizers and light of specific wavelengths. The photosensitizer is based on that the photosensitizer generates active oxygen under the irradiation of light with specific wavelength, and the active oxygen can destroy pathological tissues or pathogens, so that the aim of treatment is achieved. However, the hypoxic environment in tumor tissue may limit the efficacy of PDT. In addition, the efficiency of specific delivery of photosensitizers to tumor sites is also to be improved. Thus, there is a strong need in the art to develop complexes that target and recognize tumor cells and that are able to enhance the effect of photodynamic therapy. Disclosure of Invention The invention aims to provide a compound which can target and identify tumor cells and can enhance photodynamic therapy effect. In a first aspect of the invention, there is provided a nucleic acid nanocluster formed by agglomerating single-stranded nucleic acids, the single-stranded nucleic acids being formed by a plurality of G-quadruplex (G4) structures in series, and the single-stranded nucleic acids comprising (1) a nucleic acid aptamer AS 1411G-quadruplex (AS 1411G 4) structural unit, and (2) a human telomerase-derived G-quadruplex (Telo G) structural unit. In another preferred embodiment, the nucleic acid nanocluster comprises a structure in which the AS 1411G 4 building block and Telo G building block are sequentially repeated. In another preferred embodiment, the nucleic acid nanocluster comprises the structure of (AS 1411G 4-Telo G4) n, and subscript n represents the copy number of AS 1411G 4-Telo G4 and n is an integer of 10 or more (please review). In another preferred embodiment, the AS 1411G 4 building block and Telo G building block are connected by a linker. In another preferred embodiment, the single-stranded nucleic acid is single-stranded DNA. In another preferred embodiment, the nucleic acid nanoclusters are spherical in shape. In another preferred embodiment, the nucleic acid nanoclusters are 100-990nm in size, preferably 200-700nm, more preferably 300-600nm. In another preferred embodiment, the nucleic acid nanocluster has a polydispersity index (PDI) of 0.11±0.02. In another preferred embodiment, the nucleic acid nanoclusters are formed by rolling circle amplification from a template strand comprising (1) the complement of AS 1411G 4 and (2) the complement of Telo G4. In another preferred embodiment, the template strand has the structure shown in formula I below: Z1-G1-L-G2-Z2 (formula I) In the formula, Z1 and Z2 are primer segments; g1 is the complement of AS 1411G 4; L is none or a linker; g2 is the complement of Telo G4. In another preferred embodiment, the sequence of AS 1411G 4 is shown in SEQ ID NO. 1. In another preferred embodiment, the sequence of Telo G4 is shown as SEQ ID NO. 2. In another preferred embodiment, the L is a Poly A linker. In another preferred embodiment, the sequence of L is AAAAAAA or AAAAAAT. In another preferred embodiment, the template strand is linear. In another preferred embodiment, the linear template strand has the sequence shown in SEQ ID NO. 5. In another preferred embodiment, the template strand is a circular sequence of linear sequences joined end to end. In another preferred embodiment, the rolling circle amplification system further comprises a primer strand and a Φ29DNA polymerase. In another preferred embodiment, the 5 'end of the primer