Tumor-targeting drug carrier as well as preparation method and application thereof

Abstract

The invention discloses a tumor-targeting drug carrier, a preparation method and application of the tumor-targeting drug carrier to preparation of an antitumor drug. The antitumor targeting drug carrier is synthesized from cell penetrating peptide, bovine serum albumin and cucurbit[n]uril. The cell penetrating peptide has a cell penetrating functional zone and a shielding zone after modification,the cell penetrating functional zone is activated in a special microenvironment of tumor cell tissue, and the drug carrier is mediated to carry the drug to enter tumor cells and play an antitumor effect. Compared with exclusively used antitumor chemotherapy drugs, the drug carrier combined with the antitumor drug has good tumor cell specificity and safety and has good application prospects.

Description

technical field

[0001] The invention belongs to the technical field of biology and medicine, and in particular relates to a drug carrier with tumor-targeting effect and its preparation method and application. Background technique

[0002] Cancer is one of the malignant diseases that seriously threaten human health, and its morbidity and mortality are increasing year by year. According to estimates by the International Agency for Research on Cancer (IARC), cancer affected more than 18.1 million people worldwide in 2018, resulting in 9.6 million deaths worldwide. Timely and accurate treatment is an important means to improve the survival rate of cancer patients. As one of the main options for clinical cancer treatment, chemotherapy has achieved remarkable achievements in the past decades, with multiple anticancer drugs approved simultaneously, such as doxorubicin (DOX), oxaliplatin, and gemcitabine. However, the clinical performance of chemotherapy is poor due to the ineffic...

Examples

Embodiment 1

[0047] Example 1 Preparation and verification of Pep@BSA@CB[7]

[0048] The preparation method is as follows

[0049] A. Mix 2 μM bovine serum albumin and 40 μM cucurbit[7]uril in PBS solution and incubate at 25°C for 2 hours to form a complex BSA@CB[n] composed of cucurbit[7]uril and bovine serum albumin.

[0050] B. BSA@CB was incubated with 10 μM SPDP protein crosslinker for 2 hours in the dark, then 10 μM cell penetrating peptide Pep was added and the reaction was continued for 4 hours with stirring.

[0051] C. Centrifuge the solution in an ultrafiltration centrifuge tube (MWCO 30K) at a speed of 14,000 for 10 minutes, collect the upper unfiltered part, and dissolve it in PBS, which is Pep@BSA@CB[7].

[0052] The amino acid sequence of the polycation motif of the above cell-penetrating peptide Pep is SEQ ID NO.1, specifically R 9 GGC, the amino acid sequence of the polyanion motif is SEQ ID NO.2, specifically E 9 , the amino acid sequence of the linking motif is SEQ ID...

Embodiment 2

[0060] Example 2 Preparation and pharmacological experiments of Pep@BSA@CB[7]@DOX

[0061] 1. The preparation method is as follows

[0062] A. Mix 2 μM bovine serum albumin and 40 μM cucurbit[7]uril in PBS solution and incubate at 25°C for 2 hours to form a complex BSA@CB[7] composed of cucurbit[7]uril and bovine serum albumin.

[0063] B. Dissolve 2 μM BSA@CB[7] and 10 μM DOX in PBS (pH 7.4) and incubate for 1 hour to form a BSA@CB[7]@DOX complex, then put the solution in an ultrafiltration centrifuge tube at 14000 rpm Centrifuge for 10 minutes, collect the upper unfiltered fraction, and dissolve in PBS.

[0064] C. Incubate 2 μM BSA@CB[7]@DOX complex with 10 μM SPDP protein crosslinker for 2 hours in the dark, then add 10 μM cell penetrating peptide Pep and keep stirring for 4 hours. Centrifuge the solution in an ultrafiltration centrifuge tube at a speed of 14000 for 10 minutes, collect the upper unfiltered part, and dissolve it in PBS to obtain the Pep@BSA@CB[7] DOX comp...

technical field

[0001] The invention belongs to the technical field of biology and medicine, and in particular relates to a drug carrier with tumor-targeting effect and its preparation method and application. Background technique

[0002] Cancer is one of the malignant diseases that seriously threaten human health, and its morbidity and mortality are increasing year by year. According to estimates by the International Agency for Research on Cancer (IARC), cancer affected more than 18.1 million people worldwide in 2018, resulting in 9.6 million deaths worldwide. Timely and accurate treatment is an important means to improve the survival rate of cancer patients. As one of the main options for clinical cancer treatment, chemotherapy has achieved remarkable achievements in the past decades, with multiple anticancer drugs approved simultaneously, such as doxorubicin (DOX), oxaliplatin, and gemcitabine. However, the clinical performance of chemotherapy is poor due to the ineffic...

Examples

Embodiment 1

[0047] Example 1 Preparation and verification of Pep@BSA@CB[7]

[0048] The preparation method is as follows

[0049] A. Mix 2 μM bovine serum albumin and 40 μM cucurbit[7]uril in PBS solution and incubate at 25°C for 2 hours to form a complex BSA@CB[n] composed of cucurbit[7]uril and bovine serum albumin.

[0050] B. BSA@CB was incubated with 10 μM SPDP protein crosslinker for 2 hours in the dark, then 10 μM cell penetrating peptide Pep was added and the reaction was continued for 4 hours with stirring.

[0051] C. Centrifuge the solution in an ultrafiltration centrifuge tube (MWCO 30K) at a speed of 14,000 for 10 minutes, collect the upper unfiltered part, and dissolve it in PBS, which is Pep@BSA@CB[7].

[0052] The amino acid sequence of the polycation motif of the above cell-penetrating peptide Pep is SEQ ID NO.1, specifically R 9 GGC, the amino acid sequence of the polyanion motif is SEQ ID NO.2, specifically E 9 , the amino acid sequence of the linking motif is SEQ ID...

Embodiment 2

[0060] Example 2 Preparation and pharmacological experiments of Pep@BSA@CB[7]@DOX

[0061] 1. The preparation method is as follows

[0062] A. Mix 2 μM bovine serum albumin and 40 μM cucurbit[7]uril in PBS solution and incubate at 25°C for 2 hours to form a complex BSA@CB[7] composed of cucurbit[7]uril and bovine serum albumin.

[0063] B. Dissolve 2 μM BSA@CB[7] and 10 μM DOX in PBS (pH 7.4) and incubate for 1 hour to form a BSA@CB[7]@DOX complex, then put the solution in an ultrafiltration centrifuge tube at 14000 rpm Centrifuge for 10 minutes, collect the upper unfiltered fraction, and dissolve in PBS.

[0064] C. Incubate 2 μM BSA@CB[7]@DOX complex with 10 μM SPDP protein crosslinker for 2 hours in the dark, then add 10 μM cell penetrating peptide Pep and keep stirring for 4 hours. Centrifuge the solution in an ultrafiltration centrifuge tube at a speed of 14000 for 10 minutes, collect the upper unfiltered part, and dissolve it in PBS to obtain the Pep@BSA@CB[7] DOX comp...

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