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Liver cancer-specific gene-virus and application thereof

A specific, anti-cancer gene technology, applied in gene therapy, virus/phage, medical preparations containing active ingredients, etc., can solve problems such as non-targeting

Inactive Publication Date: 2012-11-28
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Oncolytic virus carrying gene becomes OV-gene, but Ad-gene is gene therapy (Ad has no targeting and no ability to replicate and multiply)

Method used

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  • Liver cancer-specific gene-virus and application thereof
  • Liver cancer-specific gene-virus and application thereof
  • Liver cancer-specific gene-virus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 CTGVT-LC Concrete Construction Method 1 Construction of Ad·enAFP·E1A·D55-(gene)

[0034] 1. If figure 2 as shown, Ad5(WT) It is a wild-type (i.e. normal) adenovirus 5 (Wild Type) adenovirus (Adenovirus, Ad, most of the Ad used now is type 5, so the word 5 is not specially marked), of which E1A and E1B are normal, without any modification, The market is available for sale, and our research group has reserves;

[0035] 2. ZD55 It is an oncolytic virus (Oncolytic Adenovirus, OncoAd), the gene of the 55K protein in the E1B region is deleted (deletion 55K or Δ55K, D55), and Z is the researcher's surname Zou, which is an OV that can target cancer cells , and finally dissolve it, but it has no liver cancer specificity. The construction of ZD55 is our laboratory patent (ZL 02157662.9 and ZL 200510026151.5), which has been clearly introduced, so there is no need to describe it;

[0036] 3. ZD55-(gene) (ie Ad·E1A·E1B(Δ55)-(gene)):

[0037] ZD55 is an OncoAd ve...

Embodiment 2

[0041] Embodiment 2: CTGVT-LC specific construction method 2

[0042] As mentioned above Ad·enAFP·E1A·D55-(SOCS-3), an anti-cancer gene is not effective, you can add a particularly strong anti-cancer effect, but non-tissue-specific anti-cancer gene, such as TRAIL added to the above In the same (LC) OncoAd, to form Ad·enAFP·E1A·D55-(TRAIL), the method is to insert the TRAIL expression cassette (with Bgl II cutting points at both ends) into the Bgl II site of D55, Both Ad·enAFP·E1A·D55-(SOCS-3) and Ad·enAFP·E1A·D55-(TRAIL) are shared as image 3 As shown, an excellent anticancer effect can be obtained.

[0043] In front of the two genes, different recombinants are constructed separately, and then used in combination, the anticancer effect can be greatly enhanced, because the two genes may have complementary or synergistic effects. If the two genes are connected by Linker and loaded into Ad·enAFP·E1A·D55-(TRAIL-Linker-SOCS-3), good anticancer effect can also be obtained, which ...

Embodiment 3

[0044] Embodiment 3: CTGVT-LC specific construction method 3

[0045] The enAFP promoter is used to control E1A, and the gene used is the common gene IL-24 with strong anticancer effect. Putting it into Ad·enAFP·E1A·ΔE1B also achieves unexpected anti-liver cancer effects. The cabinet frame of its construction method is as follows: Figure 4 shown.

[0046] 1. If Figure 4 as shown, Ad-Wt Same as Example 1 Medium Ad5(WT) .

[0047] 2. Construction of Ad·enAFP·E1A·ΔE1B: first use PCR to excise 19KD from D55 to form Ad·E1A·ΔE1B, and then delete E1A's natural promoter Ad·E1A(ΔNat.p) to form Ad·E1A(ΔNat .p)·ΔE1B, then add enAFP to Ad·E1A(ΔNat.p) to form Ad·enAFP·E1A·ΔE1B;

[0048] 3. Ad·enAFP·E1A·ΔE1B-(IL-24) : Ad·enAFP·E1A·ΔE1B is already a kind of oncolytic virus, which has anti-cancer effect, but not strong, such as adding an IL-24 gene expression cassette with strong anti-cancer effect, namely (IL-24), its The anti-cancer effect is stronger, and the method is to put t...

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Abstract

The invention discloses a liver cancer-specific gene-virus and application thereof. The virus is an oncolytic virus; an early gene E1 of the virus is divided into E1A and E1B; a promoter of the E1A is replaced by a liver cancer-specific promoter, and is preferably replaced by a liver cancer-specific alpha fetoprotein promoter; and an anticancer gene is carried by the virus. The anticancer gene is SOCS3, HCCS-1, TSLC-1, 1L-24, MnSOD or TRAIL. The anticancer gene is preferably IL-24. The liver cancer-specific gene-virus is used for specific medicines for treating liver cancer, has a relatively high targeting anti-hepatoma effect, basically does not affect normal cells, and also has a small effect on most of other cancer cells.

Description

technical field [0001] The present invention relates to cancer targeting gene-virus therapy (Cancer Targeting Gene-Viro-Therapy, CTGVT), in particular to liver cancer targeting gene-virus therapy (CTGVT-LC), specifically to liver cancer-specific gene-virus and its application. Background technique [0002] From 1999 to 2001, Liu Xinyuan created a cancer treatment strategy called Cancer Targeting Gene-Viro-Thearpy (CTGVT), which is to insert anti-cancer genes into oncolytic virus , OV), so the CTGVT strategy is OV-(gene) strategy or Gene Armed Oncolytic Virus Therapy (GAOVT) strategy, which is also OV-(gene), which combines the advantages of gene therapy and oncolytic virus therapy , because the oncolytic virus itself has an anti-cancer effect, and may specifically replicate hundreds of times in cancer cells, and the anti-cancer gene inserted into it may also replicate hundreds of times, so its anti-cancer effect is greatly increased. It is much better than the corresponding...

Claims

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Application Information

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IPC IPC(8): C12N7/01A61K39/235A61K48/00A61P35/00
Inventor 刘新垣韦睿成章康健曹欣
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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