Tumor ECM degrading and/or inhibiting agent and complete set kit and application thereof

A technology of inhibitors and tumors, which is applied in the direction of anti-tumor drugs, drug combinations, pharmaceutical formulations, etc., and can solve the problems of not being able to disperse and kill cancer cells

Pending Publication Date: 2019-11-05
SUZHOU KANGJU BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The actual technical problem to be solved by the present invention is to overcome that the drugs used in the prior art for treating solid tumors (including chemotherapy drugs, protein drugs, nanop

Method used

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  • Tumor ECM degrading and/or inhibiting agent and complete set kit and application thereof
  • Tumor ECM degrading and/or inhibiting agent and complete set kit and application thereof
  • Tumor ECM degrading and/or inhibiting agent and complete set kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1. Sequence Design

[0086] The amino acid sequences involved in the embodiments of the present invention include Collagenase1, PH20, HAS2 (hyaluronidase synthesis gene). The NCBI accession number of the amino acid sequence of PH20 is NP_694859.1 (cDNA is NM_153189.2); the NCBI accession number of HAS2 is NP_005319.1 (cDNA is NM_005328.3).

[0087] The amino acid sequence information of the protein used in the present invention is shown in Table 1, and the base sequence encoding the amino acid of HAS2 is shown in Table 2.

[0088] Table 1. List of amino acid sequences

[0089]

[0090]

[0091] Table 2. Base sequence list

[0092]

[0093] Genewiz Company was commissioned to synthesize the HAS2 sequence.

Embodiment 2

[0094] Example 2. Vector Construction

[0095] The base sequence of HAS2 obtained in Example 1 was digested with NheI and BamHI, ligated with DNA Ligation Kit and inserted into the corresponding site of pcDNA3.1-Hyg vector (Invitrogen). The ligation products were transformed into competent Escherichia coli (Top10). On the second day after transformation, the clones were picked and cultured in LB liquid medium containing 100 μg / mL ampicillin (Saigon Biotechnology), and sequenced for identification. The clones with correct sequencing results were selected and inoculated into LB liquid medium containing 100 μg / mL ampicillin at a scale of 50 ml and cultured for about 16 hours. Bacteria were collected, and plasmids were extracted and purified using a plasmid extraction kit (Qiagen).

[0096] The resulting HAS2-pcDNA3.1-Hyg plasmid was linearized with FspI. The digested product was extracted with phenol-chloroform to obtain a preliminary linearized plasmid. The restriction endon...

Embodiment 3

[0098] Embodiment 3. Preparation of preparation of the present invention

[0099] Different doses of hyaluronidase PH20 (Rhinobio), collagenase (Rhinobio), Losartan (Sigma) and buffer (disodium hydrogen phosphate), cryoprotectant (sucrose), excipient ( Mannitol), enzyme protection agent (which belongs to stabilizer, including disodium edetate, calcium chloride) and surfactant (Tween 20) are dissolved in water for injection, and the volume is adjusted to 6000mL. The preparation of 10,000 vials was divided into 0.6mL / vials for lyophilization.

[0100] The preparation can also be made into a solution, and the corresponding buffer (disodium hydrogen phosphate), stabilizer (sucrose, edetate disodium, calcium chloride) and surfactant (Tween 20) can be prepared according to Doses from Table 3 continued to be formulated.

[0101] Table 3. Tumor injection preparation formula

[0102]

[0103]

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PUM

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Abstract

The invention discloses a tumor ECM degrading and/or inhibiting agent and a complete set kit and application thereof. The tumor ECM degrading and/or inhibiting agent comprises at least two kinds of ananti-hyaluronidase agent, an anti-collagen agent, and an anti-tissue agent. The tumor ECM degrading and/or inhibiting agent can effectively degrade or restrain generation of a tumor extracellular matrix in a broadspectrum manner, and the tumor penetrability is improved, so that drugs can effectively enter tumors to be in contact with and kill tumor cells, and improve the medication effect. The combination manner can be applied in an intratumoral injection manner, so that the applying concentration of single components can be reduced, and the side effect is low. The complete set kit can increase the medication effect, the medicament dose can be reduced, and dosage required for achieving the same effect by an antitumor intratumoral injection manner is lower than that used for venous or other nonintratumoral injection, the antitumor dosage is reduced, and besides, the antitumous effects are increased.

Description

technical field [0001] The invention specifically relates to a tumor ECM degradation and / or inhibitor, a complete kit and application thereof. Background technique [0002] Cancer, also known as malignant tumor, is the most serious disease and persistent disease that seriously endangers human health and causes death of modern humans. According to statistics from the United Nations, one out of every eight deaths in the world is currently caused by cancer, which is higher than the number of deaths caused by AIDS, tuberculosis and malaria. Moreover, more than 10 million people around the world are diagnosed with cancer every year ("World Cancer Report 2014" released by the World Health Organization). [0003] Currently used chemotherapeutic drugs, protein drugs, nanoparticle drugs, and oncolytic virus drugs have limited therapeutic effects on solid tumors, and cell therapy methods with a high cure rate on hematological tumors have very limited effects on solid tumors. A key f...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61P35/00
CPCA61K45/06A61P35/00
Inventor 王征徐云霞
Owner SUZHOU KANGJU BIOTECHNOLOGY CO LTD
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